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NOVEL METHODS OF DIAGNOSIS OF ANGIOGENESIS, COMPOSITIONS AND METHODS
OF SCREENING FOR ANGIOGENESIS MODULATORS
FIELD OF THE INVENTION
The invention relates to the identification of expression profiles and the
nucleic acids involved in
angiogenesis, and to the use of such expression profiles and nucleic acids in
diagnosis of
angiogenesis. The invention further relates to methods for identifying
candidate agents and/or targets
which modulate angiogenesis.
BACKGROUND OF THE INVENTION
New blood vessel development comprises the formation of veins (vasculogenesis)
and arteries
(angiogenesis). Angiogenesis plays a normal role in embryonic development, as
well as menstration,
wound healing. Angiogenesis also plays a crucial pathogenic role in a variety
of disease states,
including cancer, proliferative diabetic retinopathy, and maintaining blood
flow to chronic inflammatory
sites.
Angiogenesis has a number of stages. The early stages of angiogenesis include
endothelial cell
protease production, migration of cells and proliferation. The early stages
also appear to require some
growth factors, with VEGF, TGF-a, angiostatin, and selected chemokines all
putatively playing a role.
Later stages of angiogenesis include the population of the vessels with mural
cells (pericytes or
smooth muscle cells), basement membrane production and the induction of vessel
bed
specializations. The final stages of vessel formation include what is known as
"remodeling", wherein a
2 0 forming vasculature becomes a stable, mature vessel bed.
Thus, understanding the genes, proteins and regulatory mechanisms that occur
during angiogenesis
would be desirable. Accordingly, it is an object of the invention to provide
methods that can be used to
screen candidate bioactive agents for the ability to modulate angiogenesis.
Additionally, it is an object
to provide molecular targets for therapeutic intervention in disease states
which either have an
undesirable excess or a deficit in angiogenesis.
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SUMMARY OF THE INVENTION
The present invention provides novel methods for diagnosis and prognosis
evaluation for
angiogenesis, as well as methods for screening for compositions which modulate
angiogenesis.
Methods of treatment of disorders associated with angiogenesis, as well as
compositions are also
provided herein.
In one aspect, a method of screening drug candidates comprises providing a
cell that expresses an
expression profile gene or fragments thereof. or fragments thereof. Preferred
embodiments of the
expression profile gene are genes which are differentially expressed in
angiogenesis cells, compared
to other cells. Preferred embodiments of expression profile genes used in the
methods herein include
but are not limited to the group consisting of AAA4, AAA1, Edg-1, alpha 5
beta1 integrin, endomucin
and matrix metalloproteinase 10; fragments of the proteins of this group are
also preferred. It is
understood that molecules for use in the present invention may be from any
figure or any subset of
listed molecules. Therefore, for example, any one or more of the genes listed
above can be used in
the methods herein. In another embodiment, a nucleic acid is selected from
Tables 1, 2, 3, 4 or 5.
Preferred nucleic acids are in Table 4, and most preferably Table 5. The
method further includes
adding a drug candidate to the cell and determining the effect of the drug
candidate on the expression
of the expression profile gene.
In one embodiment, the method of screening drug candidates includes comparing
the level of
expression in the absence of the drug candidate to the level of expression in
the presence of the drug
2 0 candidate, wherein the concentration of the drug candidate can vary when
present, and wherein the
comparison can occur after addition or removal of the drug candidate. In a
preferred embodiment, the
cell expresses at least two expression profile genes. The profile genes may
show an increase or
decrease.
Also provided herein is a method of screening for a bioactive agent capable of
binding to an
angiogenesis modulator protein (AMP), the method comprising combining the AMP
and a candidate
bioactive agent, and determining the binding of the candidate agent to the
AMP. Preferably the AMP
is a protein or fragment thereof selected from the group consisting of AAA4,
AAA1, Edg-1, alpha 5
beta1 integrin, endomucin and matrix metalloproteinase 10. In another
embodiment, the proteins is
encoded by a nucleic acid selected from Tables 1, 2, 3, 4 or 5. Preferred
nucleic acids are in Table 4,
3 0 and most preferably Table 5.
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Further provided herein is a method for screening for a bioactive agent
capable of modulating the
activity of an AMP. In one embodiment the method comprises combining the AMP
and a candidate
bioactive agent, and determining the effect of the candidate agent on the
bioactivity of the AMP.
Preferably the AMP is a protein or fragment thereof selected from the group
consisting of AAA4,
AAA1, Edg-1, alpha 5 beta1 integrin, endomucin and matrix metalloproteinase
10. In another
embodiment, the proteins is encoded by a nucleic acid selected from Tables 1,
2, 3, 4 or 5. Preferred
nucleic acids are in Table 4, and most preferably Table 5.
Also provided is a method of evaluating the effect of a candidate angiogenesis
drug comprising
administering the drug to a transgenic animal expressing or over-expressing
the AMP, or an animal
lacking the AMP, for example as a result of a gene knockout.
Additionally, provided herein is a method of evaluating the effect of a
candidate angiogenesis drug
comprising administering the drug to a patient and removing a cell sample from
the patient. The
expression profile of the cell is then determined. This method may further
comprise comparing the
expression profile to an expression profile of a healthy individual. In a
preferred embodiment, the
expression profile includes a gene of Table 1, Table 2, Table 3, Table 4 or
Table 5.
Moreover, provided herein is a biochip comprising one or more nucleic acid
segments which encode
an angiogenesis protein, preferable selected from the group consisting of
AAA4, AAA1, Edg-1, alpha 5
beta1 integrin, endomucin and matrix metalloproteinase , or fragment thereof,
wherein the biochip
comprises fewer than 1000 nucleic acid probes. Preferably at least two nucleic
acid segments are
2 0 included. In another embodiment, the nucleic acid selected from Tables 1,
2, 3, 4 or 5. Preferred
nucleic acids are in Table 4, and most preferably Table 5.
Furthermore, a method of diagnosing a disorder associated with angiogenesis is
provided. The
method comprises determining the expression of a gene which encodes an
angiogenesis protein
preferable selected from the group consisting of AAA4, AAA1, Edg-1, alpha 5
beta1 integrin,
2 5 endomucin and matrix metalloproteinase 10, or fragment thereof in a first
tissue type of a first
individual, and comparing the distribution to the expression of the gene from
a second normal tissue
type from the first individual or a second unaffected individual. In another
embodiment, the proteins is
encoded by a nucleic acid selected from Tables 1, 2, 3, 4 or 5. Preferred
nucleic acids are in Table 4,
and most preferably Table 5. A difference in the expression indicates that the
first individual has a
3 0 disorder associated with angiogenesis.
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In another aspect, the present invention provides an antibody which
specifically binds to an
angiogenesis preferably selected from the group consisting of AAA4, AAA1, Edg-
1, alpha 5 beta1
integrin, endomucin and matrix metalloproteinase 10 or fragment thereof. . In
another embodiment,
the proteins is encoded by a nucleic acid selected from Tables 1, 2, 3, 4 or
5. Preferred nucleic acids
are in Table 4, and most preferably Table 5. In a preferred embodiment the
fragment of AAA1 is
selected from AAA1 p1 or AAA1 p2. Other preferred fragments for the
angiogenesis proteins are
shown in the figures.
In one embodiment a method for screening for a bioactive agent capable of
interfering with the binding
of a angiogenesis modulating protein (AMP) or a fragment thereof and an
antibody which binds to said
AMP or fragment thereof. In a preferred embodiment, the method comprises
combining an AMP or
fragment thereof, a candidate bioactive agent and an antibody which binds to
said AMP or fragment
thereof. The method further includes determining the binding of said AMP or
fragment thereof and
said antibody. Wherein there is a change in binding, an agent is identified as
an interfering agent.
The interfering agent can be an agonist or an antagonist. Preferably, the
agent inhibits angiogenesis.
In a further aspect, a method for inhibiting angiogenesis is provided. In one
embodiment, the method
comprises administering to a cell a composition comprising an antibody to an
angiogenesis
modulating protein, preferably selected from the group consisting of AAA4,
AAA1, Edg-1, alpha 5
beta1 integrin, endomucin and matrix metalloproteinase 10, or fragment
thereof. In another
embodiment, the proteins is encoded by a nucleic acid selected from Tables 1,
2, 3, 4 or 5. Preferred
2 0 nucleic acids are in Table 4, and most preferably Table 5. The method can
be performed in vitro or in
vivo, preferably in vivo to an individual. In a preferred embodiment the
method of inhibiting
angiogenesis is provided to an individual with a disorder associated with
angiogenesis such as cancer.
As described herein, methods of inhibiting angiogenesis can be performed by
administering an
inhibitor of the activity of an angiogenesis protein, including an antisense
molecule to the gene or its
2 5 gene products, and preferable small molecules.
Also provided herein are methods of eliciting an immune response in an
individual. In one
embodiment a method provided herein comprises administering to an individual a
composition
comprising an angiogenesis modulating protein, preferably selected from the
group consisting of
AAA4, AAA1, Edg-1, alpha 5 beta1 integrin, endomucin and matrix
metalloproteinase 10, or fragment
3 0 thereof. In another embodiment, the proteins is encoded by a nucleic acid
selected from Tables 1, 2,
3, 4 or 5. Preferred nucleic acids are in Table 4, and most preferably Table
5. In another aspect, said
composition comprises a nucleic acid comprising a sequence encoding an
angiogenesis modulating
protein, preferably selected from the group consisting of AAA4, AAA1, Edg-1,
alpha 5 beta1 integrin,
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endomucin and matrix metalloproteinase 10, or fragment thereof. In another
embodiment, the
proteins is encoded by a nucleic acid selected from Tables 1, 2, 3, 4 or 5.
Preferred nucleic acids are
in Table 4, and most preferably Table 5.
Further provided herein are compositions capable of eliciting an immune
response in an individual. In
one embodiment, a composition provided herein comprises an angiogenesis
modulating protein,
preferably selected from the group consisting of AAA4, AAA1, Edg-1, alpha 5
beta1 integrin,
endomucin and matrix metalloproteinase 10, or fragment thereof. In another
embodiment, the
proteins is encoded by a nucleic acid selected from Tables 1, 2, 3, 4 or 5.
Preferred nucleic acids are
in Table 4, and most preferably Table 5. In another embodiment, said
composition comprises a
nucleic acid comprising a sequence encoding an angiogenesis modulating
protein, preferably selected
from the group consisting of AAA4, AAA1, Edg-1, alpha 5 beta1 integrin,
endomucin and matrix
metalloproteinase 10, or fragment thereof, and a pharmaceutically acceptable
carrier.
In another embodiment the nucleic acid selected from Tables 1, 2, 3, 4 or 5.
Preferred nucleic acids
are in Table 4, and most preferably Table 5.
A method of neutralizing the effect of an angiogenesis protein, preferably
selected from the group
consisting of AAA4, AAA1, Edg-1, alpha 5 beta1 integrin, endomucin and matrix
metalloproteinase 10,
or fragment thereof, comprising contacting an agent specific for said protein
with said protein in an
amount sufficient to effect neutralization. In another embodiment, the
proteins is encoded by a nucleic
acid selected from Tables 1, 2, 3, 4 or 5. Preferred nucleic acids are in
Table 4, and most preferably
2 0 Table 5.
In another aspect of the invention, a method of treating an individual for a
disorder associated with
angiogenesis is provided. In one embodiment, the method comprises
administering to said individual
an inhibitor of Edg-1. In another embodiment, the method comprises
administering to a patient having
a disorder with angiogenesis an antibody to Edg-1 conjugated to a therapeutic
moiety. Such a
2 S therapeutic moiety can be a cytotoxic agent or a radioisotope.
Novel sequences are provided herein. Compounds and compositions are also
provided. Other
aspects of the invention will become apparent to the skilled artisan by the
following description of the
invention.
DETAILED DESCRIPTION OF THE TABLES AND FIGURES
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Table 1 provides the Accession numbers for 1774 genes, including expression
sequence tags,
(incorporated in their entirety here and throughout the application where
Accession numbers are
provided), whose expression levels change as a function of time in tissue
undergoing angiogenesis
compared to tissue that is not.
Table 2 provides the Accession numbers for a preferred subset of 559 genes,
including expression
sequence tags (incorporated in their entirety here and throughout the
application where Accession
numbers are provided), whose expression levels change as a function of time in
tissue undergoing
angiogenesis compared to tissue that is not. The sequences are characterized
as predicted to
encode secreted proteins (SS), or transmembrane proteins (TM) proteins.
Table 3 provides the Accession numbers for 1916 genes including expression
sequence tags
(incorporated in their entirety here and throughout the application where
Accession numbers are
provided), whose expression levels change as a function of time in tissue
undergoing angiogenesis
compared to tissue that is not.
Table 4 provides a preferred subset of 558 Accession numbers identified in
Figure 4 whose
expression levels change as a function of time in tissue undergoing
angiogenesis compared to tissue
that is not.
Table 5 provides a preferred subset of 20 Accession numbers identified in
Figure 4 whose expression
levels change as a function of time in tissue undergoing angiogenesis compared
to tissue that is not.
Figure 1 is a graph of expression levels of sequences identified in Figure 1.
Expression profiles are
2 0 clustered into 4 groups. C1 (blue), C2 (red), C3 (green) and C4 (mustard).
Figure 2 shows an embodiment of a nucleic acid (mRNA) which includes a
sequence encoding an
angiogenesis protein, AAA4. The start and stop codons are underlined.
Figure 3 shows the open reading frame of a nucleic acid sequence encoding
AAA4. The start and
stop codons are underlined.
Figure 4 shows an embodiment of the amino acid sequence of AAA4. The signal
peptide is double
underlined, and the transmembrane sequence is underlined. In one embodiment
herein, AAA4 is
soluble. Thus, the signal peptide can be omitted, and the transmembrane domain
deleted,
inactivated, or truncated.
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Figure 5 shows peptides AAA4p1 and AAA4p2.
Figure 6 shows the expression of AAA4 in angiogenesis models over time and in
other, non-
angiogenic tissues.
Figure 7 shows an embodiment of a nucleic acid sequence encoding an
angiogenesis protein, AAA1.
A putative stop codon is underlined.
Figure 8 shows an embodiment of an amino acid sequence for AAA1. A
transmembrane domain is
underlined. In one embodiment, AAA1 is soluble. In preferred embodiments, the
transmembrane
domain is deleted or inactivated, or AAA1 is truncated to delete the
transmembrane domain.
Figure 9 shows AAA1 p1 and AAA1 p2.
Figure 10 shows a graph showing the relative expression of AAA1 in various
tissues at different time
points. "Exp 3" is an angiogenesis model showing tube formation over time
using endothelial cells.
Figure 11 shows an embodiment of a nucleic acid, mRNA, which comprises a
sequence encoding an
angiogenesis protein, Edg-1. The start and stop codons are underlined.
Figure 12 shows the open reading frame encoding Edg-1, wherein the start and
stop codons are
underlined.
Figure 13 shows an embodiment of an amino acid sequence for an angiogenesis
protein, Edg-1,
wherein the transmembrane domains are underlined. In a preferred embodiment
herein, a soluble
2 0 form of Edg-1 is provided. In one embodiment, the transmembrane domains
are deleted, inactivated,
and/or the protein is truncated so as to exclude the domains (with or without
re-ligation of remaining
soluble regions).
Figure 14 depicts four peptide sequences provided herein and their respective
solubilities.
Figure 15 shows the expression of Edg-1 over a variety of tissues.
Figure 16 shows the time course of induction of Edg-1 in a model for
angiogenesis (Expt 1, Expt 2,
Expt 3) in which low passage human endothelial cells form into tube structures
over a period of a few
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days in culture. The reproducible induction of Edg-1 occurred in a time frame
consistent with its role in
the tube forming process.
Figure 17 shows an embodiment of a nucleic acid sequence which includes the
coding sequence for a
tissue remodeling protein, alpha 5 beta 1 integrin (sometimes referred to as
VLA-5), wherein the start
and stop codon are underlined.
Figure 18 shows an embodiment of an amino acid sequence of a tissue remodeling
protein, alpha 5
beta 1 integrin, wherein a transmembrane domain is underlined.
Figure 19 shows a bar graph depicting the results of 5 expression profiles of
alpha 5 beta 1 integrin
throughout the time course of tube formation. In particular, tube models 1, 2
and 3 show models
which form tube structures from single isolated human endothelial cells; the
"EC/PMA" model shows
endothelial cells stimulated with pokeweed mitogen antigen, and the body atlas
profile shows
expression in various normal cell types and tissues.
Figures 20A and 20B show the results of antagonism of tube formation wherein
Figure 20A is an
isotype control and Figure 20B shows specific antibody antagonism after 48
hours.
Figure 21 shows an embodiment of a nucleic acid sequence which includes the
coding sequence for
an angiogenesis protein, endomucin, wherein the start and stop codon are
boxed.
Figure 22 shows an embodiment of an amino acid sequence of an angiogenesis
protein, endomucin,
wherein a signal sequence is bolded and a transmembrane domain is underlined.
Figure 23 shows an embodiment of a nucleic acid sequence which includes the
coding sequence for
2 0 an angiogenesis protein, matrix metalloproteinase 10 (also called
stromolysin 2), wherein the start and
stop codon are boxed.
Figure 24 shows expression of matrix metalloproteinase 10 over a variety of
tissues.
Figure 25 shows expression of matrix metalloproteinase 10 over a variety of
tissues.
DETAILED DESCRIPTION OF THE INVENTION
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In accordance with the objects outlined above, the present invention provides
novel methods for
diagnosis of disorders associated with angiogenesis (sometimes referred to
herein as angiogenesis
disorders or AD), as well as methods for screening for compositions which
modulate angiogenesis.
By "disorder associated with angiogenesis" or "disease associated with
angiogenesis" herein is meant
a disease state which is marked by either an excess or a deficit of vessel
development. Angiogenesis
disorders include, but are not limited to, cancer and proliferative diabetic
retinopathy. Also provided
are method for treating AD.
In one aspect, the expression levels of genes are determined in different
patient samples for which
diagnosis information is desired, to provide expression profiles. An
expression profile of a particular
sample is essentially a "fingerprint" of the state of the sample; while two
states may have any
particular gene similarly expressed, the evaluation of a number of genes
simultaneously allows the
generation of a gene expression profile that is unique to the state of the
cell. That is, normal tissue
may be distinguished from AD tissue. By comparing expression profiles of
tissue in known different
angiogenesis states, information regarding which genes are important
(including both up- and down-
regulation of genes) in each of these states is obtained. The identification
of sequences that are
differentially expressed in angiogenic versus non-angiogenic tissue allows the
use of this information
in a number of ways. For example, the evaluation of a particular treatment
regime may be evaluated:
does a chemotherapeutic drug act to down-regulate angiogenesis and thus tumor
growth or
recurrence in a particular patient. Similarly, diagnosis may be done or
confirmed by comparing patient
2 0 samples with the known expression profiles. Furthermore, these gene
expression profiles (or
individual genes) allow screening of drug candidates with an eye to mimicking
or altering a particular
expression profile; for example, screening can be done for drugs that suppress
the angiogenic
expression profile. This may be done by making biochips comprising sets of the
important
angiogenesis genes, which can then be used in these screens. These methods can
also be done on
2 5 the protein basis; that is, protein expression levels of the angiogenic
proteins can be evaluated for
diagnostic purposes or to screen candidate agents. In addition, the angiogenic
nucleic acid
sequences can be administered for gene therapy purposes, including the
administration of antisense
nucleic acids, or the angiogenic proteins (including antibodies and other
modulators thereof)
administered as therapeutic drugs.
3 0 Thus the present invention provides nucleic acid and protein sequences
that are differentially
expressed in angiogenesis, herein termed "angiogenesis sequences". As outlined
below,
angiogenesis sequences include those that are up-regulated (i.e. expressed at
a higher level) in
disorders associated with angiogenesis, as well as those that are down-
regulated (i.e. expressed at a
lower level). In a preferred embodiment, the angiogenesis sequences are from
humans; however, as
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will be appreciated by those in the art, angiogenesis sequences from other
organisms may be useful
in animal models of disease and drug evaluation; thus, other angiogenesis
sequences are provided,
from vertebrates, including mammals, including rodents (rats, mice, hamsters,
guinea pigs, etc.),
primates, farm animals (including sheep, goats, pigs, cows, horses, etc).
Angiogenesis sequences
from other organisms may be obtained using the techniques outlined below.
Angiogenesis sequences can include both nucleic acid and amino acid sequences.
In a preferred
embodiment, the angiogenesis sequences are recombinant nucleic acids. By the
term "recombinant
nucleic acid" herein is meant nucleic acid, originally formed in vitro, in
general, by the manipulation of
nucleic acid by polymerases and endonucleases, in a form not normally found in
nature. Thus an
isolated nucleic acid, in a linear form, or an expression vector formed in
vitro by ligating DNA
molecules that are not normally joined, are both considered recombinant for
the purposes of this
invention. It is understood that once a recombinant nucleic acid is made and
reintroduced into a host
cell or organism, it will replicate non-recombinantly, i.e. using the in vivo
cellular machinery of the host
cell rather than in vitro manipulations; however, such nucleic acids, once
produced recombinantly,
although subsequently replicated non-recombinantly, are still considered
recombinant for the purposes
of the invention.
Similarly, a "recombinant protein" is a protein made using recombinant
techniques, i.e. through the
expression of a recombinant nucleic acid as depicted above. A recombinant
protein is distinguished
from naturally occurring protein by at least one or more characteristics. For
example, the protein may
2 0 be isolated or purified away from some or all of the proteins and
compounds with which it is normally
associated in its wild type host, and thus may be substantially pure. For
example, an isolated protein
is unaccompanied by at least some of the material with which it is normally
associated in its natural
state, preferably constituting at least about 0.5%, more preferably at least
about 5% by weight of the
total protein in a given sample. A substantially pure protein comprises at
least about 75% by weight of
2 5 the total protein, with at least about 80% being preferred, and at least
about 90% being particularly
preferred. The definition includes the production of an angiogenesis protein
from one organism in a
different organism or host cell. Alternatively, the protein may be made at a
significantly higher
concentration than is normally seen, through the use of an inducible promoter
or high expression
promoter, such that the protein is made at increased concentration levels.
Alternatively, the protein
3 0 may be in a form not normally found in nature, as in the addition of an
epitope tag or amino acid
substitutions, insertions and deletions, as discussed below.
In a preferred embodiment, the angiogenesis sequences are nucleic acids. As
will be appreciated by
those in the art and is more fully outlined below, angiogenesis sequences are
useful in a variety of
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applications, including diagnostic applications, which will detect naturally
occurring nucleic acids, as
well as screening applications; for example, biochips comprising nucleic acid
probes to the
angiogenesis sequences can be generated. In the broadest sense, then, by
"nucleic acid" or
"oligonucleotide" or grammatical equivalents herein means at least two
nucleotides covalently linked
together. A nucleic acid of the present invention will generally contain
phosphodiester bonds, although
in some cases, as outlined below, nucleic acid analogs are included that may
have alternate
backbones, comprising, for example, phosphoramidate (Beaucage et al.,
Tetrahedron 49(10):1925
(1993) and references therein; Letsinger, J. Org. Chem. 35:3800 (1970);
Sprinzl et al., Eur. J.
Biochem. 81:579 (1977); Letsinger et al., Nucl. Acids Res. 14:3487 (1986);
Sawai et al, Chem. Lett.
805 (1984), Letsinger et al., J. Am. Chem. Soc. 110:4470 (1988); and Pauwels
et al., Chemica Scripts
26:141 91986)), phosphorothioate (Mag et al., Nucleic Acids Res. 19:1437 (1991
); and U.S. Patent
No. 5,644,048), phosphorodithioate (Briu et al., J. Am. Chem. Soc. 111:2321
(1989), O-
methylphophoroamidite linkages (see Eckstein, Oligonucleotides and Analogues:
A Practical
Approach, Oxford University Press), and peptide nucleic acid backbones and
linkages (see Egholm, J.
Am. Chem. Soc. 114:1895 (1992); Meier et al., Chem. Int. Ed. Engl. 31:1008
(1992); Nielsen, Nature,
365:566 (1993); Carlsson et al., Nature 380:207 (1996), all of which are
incorporated by reference).
Other analog nucleic acids include those with positive backbones (Denpcy et
al., Proc. Natl. Acad. Sci.
USA 92:6097 (1995); non-ionic backbones (U.S. Patent Nos. 5,386,023,
5,637,684, 5,602,240,
5,216,141 and 4,469,863; Kiedrowshi et al., Angew. Chem. Intl. Ed. English
30:423 (1991); Letsinger
2 0 et al., J. Am. Chem. Soc. 110:4470 (1988); Letsinger et al., Nucleoside &
Nucleotide 13:1597 (1994);
Chapters 2 and 3, ASC Symposium Series 580, "Carbohydrate Modifications in
Antisense Research",
Ed. Y.S. Sanghui and P. Dan Cook; Mesmaeker et al., Bioorganic & Medicinal
Chem. Lett. 4:395
(1994); Jeffs et al., J. Biomolecular NMR 34:17 (1994); Tetrahedron Lett.
37:743 (1996)) and non-
ribose backbones, including those described in U.S. Patent Nos. 5,235,033 and
5,034,506, and
2 5 Chapters 6 and 7, ASC Symposium Series 580, "Carbohydrate Modifications in
Antisense Research",
Ed. Y.S. Sanghui and P. Dan Cook. Nucleic acids containing one or more
carbocyclic sugars are also
included within one definition of nucleic acids (see Jenkins et al., Chem.
Soc. Rev. (1995) pp169-
176). Several nucleic acid analogs are described in Rawls, C & E News June 2,
1997 page 35. All of
these references are hereby expressly incorporated by reference. These
modifications of the ribose-
3 0 phosphate backbone may be done for a variety of reasons, for example to
increase the stability and
half-life of such molecules in physiological environments or as probes on a
biochip.
As will be appreciated by those in the art, all of these nucleic acid analogs
may find use in the present
invention. In addition, mixtures of naturally occurring nucleic acids and
analogs can be made;
alternatively, mixtures of different nucleic acid analogs, and mixtures of
naturally occurring nucleic
3 5 acids and analogs may be made.
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Particularly preferred are peptide nucleic acids (PNA) which includes peptide
nucleic acid analogs.
These backbones are substantially non-ionic under neutral conditions, in
contrast to the highly
charged phosphodiester backbone of naturally occurring nucleic acids. This
results in two
advantages. First, the PNA backbone exhibits improved hybridization kinetics.
PNAs have larger
changes in the melting temperature (Tm) for mismatched versus perfectly
matched basepairs. DNA
and RNA typically exhibit a 2-4°C drop in Tm for an internal mismatch.
With the non-ionic PNA
backbone, the drop is closer to 7-9°C. Similarly, due to their non-
ionic nature, hybridization of the
bases attached to these backbones is relatively insensitive to salt
concentration. In addition, PNAs
are not degraded by cellular enzymes, and thus can be more stable.
The nucleic acids may be single stranded or double stranded, as specified, or
contain portions of both
double stranded or single stranded sequence. As will be appreciated by those
in the art, the depiction
of a single strand ("Watson") also defines the sequence of the other strand
("Crick"); thus the
sequences described herein also includes the complement of the sequence. The
nucleic acid may be
DNA, both genomic and cDNA, RNA or a hybrid, where the nucleic acid contains
any combination of
deoxyribo- and ribo-nucleotides, and any combination of bases, including
uracil, adenine, thymine,
cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine, isoguanine,
etc. As used herein, the
term "nucleoside" includes nucleotides and nucleoside and nucleotide analogs,
and modified
nucleosides such as amino modified nucleosides. In addition, "nucleoside"
includes non-naturally
occurring analog structures. Thus for example the individual units of a
peptide nucleic acid, each
2 0 containing a base, are referred to herein as a nucleoside.
An angiogenesis sequence can be initially identified by substantial nucleic
acid and/or amino acid
sequence homology to the angiogenesis sequences outlined herein. Such homology
can be based
upon the overall nucleic acid or amino acid sequence, and is generally
determined as outlined below,
using either homology programs or hybridization conditions.
2 5 The angiogenesis screen included comparing genes identified in an in vitro
model of angiogenesis as
described in Hiraoka, Cell 95:365 (1998), which is expressly incorporated by
reference, with genes
identified in controls. Samples of normal tissue and tissue undergoing
angiogenesis are applied to
biochips comprising nucleic acid probes. The samples are first microdissected,
if applicable, and
treated as is known in the art for the preparation of mRNA. Suitable biochips
are commercially
3 0 available, for example from Affymetrix. Gene expression profiles as
described herein are generated
and the data analyzed.
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In a preferred embodiment, the genes showing changes in expression as between
normal and
disease states are compared to genes expressed in other normal tissues,
including, but not limited to
lung, heart, brain, liver, breast, kidney, muscle, prostate, small intestine,
large intestine, spleen, bone
and placenta. In a preferred embodiment, those genes identified during the
angiogenesis screen that
are expressed in any significant amount in other tissues are removed from the
profile, although in
some embodiments, this is not necessary. That is, when screening for drugs, it
is preferable that the
target be disease specific, to minimize possible side effects.
In a preferred embodiment, angiogenesis sequences are those that are up-
regulated in angiogenesis
disorders; that is, the expression of these genes is higher in the disease
tissue as compared to normal
tissue. "Up-regulation" as used herein means at least about a two-fold change,
preferably at least
about a three fold change, with at least about five-fold or higher being
preferred. All accession
numbers herein are for the GenBank sequence database and the sequences of the
accession
numbers are hereby expressly incorporated by reference. GenBank is known in
the art, see, e.g.,
Benson, DA, et al., Nucleic Acids Research 26:1-7 (1998) and
http://www.ncbi.nlm.nih.gov/. In
addition, these genes were found to be expressed in a limited amount or not at
all in heart, brain, lung,
liver, breast, kidney, prostate, small intestine and spleen.
In a preferred embodiment, angiogenesis sequences are those that are down-
regulated in the
angiogenesis disorder; that is, the expression of these genes is lower in
angiogenic tissue as
compared to normal tissue. "Down-regulation" as used herein means at least
about a two-fold change,
2 0 preferably at least about a three fold change, with at least about five-
fold or higher being preferred.
Angiogenesis sequences according to the invention may be classified into
discrete clusters of
sequences based on common expression profiles of the sequences. Expression
levels of
angiogenesis sequences may increase or decrease as a function of time in a
manner that correlates
with the induction of angiogenesis. Alternatively, expression levels of
angiogenesis sequences may
2 5 both increase and decrease as a function of time. For example, expression
levels of some
angiogenesis sequences are temporarily induced or diminished during the switch
to the angiogenesis
phenotype, followed by a return to baseline expression levels. Table 1 depicts
1774 genes, the
expression of which varies as a function of time in angiogenesis tissue when
compared to normal
tissue. Figure 1 depicts 4 discrete expression profiles of angiogenesis genes
identified in Table 1.
3 0 A particularly preferred embodiment includes the sequences as described in
Table 2 which depicts a
preferred subset of 559 angiogenesis sequences, the expression of which is
altered in angiogenesis
when compared to normal tissue.
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An additional embodiment includes the sequences as described in Table 3, which
depicts 1916 genes
including expression sequence tags (incorporated in their entirety here and
throughout the application
where Accession numbers are provided), whose expression levels change as a
function of time in
tissue undergoing angiogenesis compared to tissue that is not.
A preferred embodiment includes the sequences as described in Table 4 which
depicts a preferred
subset of 558 genes identified in Table 3 whose expression levels change as a
function of time in
tissue undergoing angiogenesis compared to tissue that is not.
A particularly preferred embodiment includes the sequences as described in
Table 5 which provides a
preferred subset of 20 Accession numbers identified in Table 3 whose
expression levels change as a
function of time in tissue undergoing angiogenesis compared to tissue that is
not.
In a particularly preferred embodiment, angiogenesis sequences are those that
are induced for a
period of time followed by a return to the baseline levels. Sequences that are
temporarily induced
provide a means to target angiogenesis tissue, for example neovascularized
tumors, while avoiding
rapidly growing tissue that require perpetual vascularization. Such positive
angiogenic factors include
aFGF, bFGF, VEGF, angiogenin and the like.
Induced angiogenesis sequences also are further categorized with respect to
the timing of induction.
For example, some angiogenesis genes may be induced at an early time period,
such as with 10
minutes of the induction of angiogenesis. Others may be induced later, such as
between 5 and 60
minutes, while yet others may be induced for a time period of about two hours
or more followed by a
2 0 return to baseline expression levels.
In another preferred embodiment are angiogenesis sequences that are inhibited
or reduced as a
function of time followed by a return to "normal" expression levels.
Inhibitors of angiogenesis are
examples of molecules that have this expression profile. These sequences also
can be further divided
into groups depending on the timing of diminished expression. For example,
some molecules may
2 5 display reduced expression with 10 minutes of the induction of
angiogenesis. Others may be
diminished later, such as between 5 and 60 minutes, while others may be
diminished for a time period
of about two hours or more followed by a return to baseline. Examples of such
negative angiogenic
factors include thrombospondin and endostatin to name a few.
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In yet another preferred embodiment are angiogenesis sequences that are
induced for prolonged
periods. These sequences are typically associated with induction of
angiogenesis and may participate
in induction and/or maintenance of the angiogenesis phenotype.
In another preferred embodiment are angiogenesis sequences, the expression of
which is reduced or
diminished for prolonged periods in angiogenic tissue. These sequences are
typically angiogenesis
inhibitors and their diminution is correlated with an increase in
angiogenesis.
Angiogenesis proteins of the present invention may be classified as secreted
proteins,
transmembrane proteins or intracellular proteins. In a preferred embodiment
the angiogenesis protein
is an intracellular protein. Intracellular proteins may be found in the
cytoplasm and/or in the nucleos.
Intracellular proteins are involved in all aspects of cellular function and
replication (including, for
example, signaling pathways); aberrant expression of such proteins results in
unregulated or
disregulated cellular processes. For example, many intracellular proteins have
enzymatic activity such
as protein kinase activity, protein phosphatase activity, protease activity,
nucleotide cyclase activity,
polymerase activity and the like. Intracellular proteins also serve as docking
proteins that are involved
in organizing complexes of proteins, or targeting proteins to various
subcellular localizations, and are
involved in maintaining the structural integrity of organelles.
An increasingly appreciated concept in characterizing intracellular proteins
is the presence in the
proteins of one or more motifs for which defined functions have been
attributed. In addition to the
highly conserved sequences found in the enzymatic domain of proteins, highly
conserved sequences
2 0 have been identified in proteins that are involved in protein-protein
interaction. For example, Src-
homology-2 (SH2) domains bind tyrosine-phosphorylated targets in a sequence
dependent manner.
PTB domains, which are distinct from SH2 domains, also bind tyrosine
phosphorylated targets. SH3
domains bind to proline-rich targets. In addition, PH domains,
tetratricopeptide repeats and WD
domains to name only a few, have been shown to mediate protein-protein
interactions. Some of these
may also be involved in binding to phospholipids or other second messengers.
As will be appreciated
by one of ordinary skill in the art, these motifs can be identified on the
basis of primary sequence;
thus, an analysis of the sequence of proteins may provide insight into both
the enzymatic potential of
the molecule and/or molecules with which the protein may associate.
In a preferred embodiment, the angiogenesis sequences are transmembrane
proteins.
3 0 Transmembrane proteins are molecules that span the phospholipid bilayer of
a cell. They may have
an intracellular domain, an extracellular domain, or both. The intracellular
domains of such proteins
may have a number of functions including those already described for
intracellular proteins. For
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example, the intracellular domain may have enzymatic activity and/or may serve
as a binding site for
additional proteins. Frequently the intracellular domain of transmembrane
proteins serves both roles.
For example certain receptor tyrosine kinases have both protein kinase
activity and SH2 domains. In
addition, autophosphorylation of tyrosines on the receptor molecule itself,
creates binding sites for
additional SH2 domain containing proteins.
Transmembrane proteins may contain from one to many transmembrane domains. For
example,
receptor tyrosine kinases, certain cytokine receptors, receptor guanylyl
cyclases and receptor
serine/threonine protein kinases contain a single transmembrane domain.
However, various other
proteins including channels and adenylyl cyclases contain numerous
transmembrane domains. Many
important cell surface receptors are classified as "seven transmembrane
domain" proteins, as they
contain 7 membrane spanning regions. Important transmembrane protein receptors
include, but are
not limited to insulin receptor, insulin-like growth factor receptor, human
growth hormone receptor,
glucose transporters, transferrin receptor, epidermal growth factor receptor,
low density lipoprotein
receptor, epidermal growth factor receptor, leptin receptor, interleukin
receptors, e.g. IL-1 receptor,
IL-2 receptor, etc.
Characteristics of transmembrane domains include approximately 20 consecutive
hydrophobic amino
acids that may be followed by charged amino acids. Therefore, upon analysis of
the amino acid
sequence of a particular protein, the localization and number of transmembrane
domains within the
protein may be predicted.
2 0 The extracellular domains of transmembrane proteins are diverse; however,
conserved motifs are
found repeatedly among various extracellular domains. Conserved structure
and/or functions have
been ascribed to different extracellular motifs. For example, cytokine
receptors are characterized by a
cluster of cysteines and a WSXWS (W= tryptophan, S= serine, X=any amino acid)
motif.
Immunoglobulin-like domains are highly conserved. Mucin-like domains may be
involved in cell
2 5 adhesion and leucine-rich repeats participate in protein-protein
interactions.
Many extracellular domains are involved in binding to other molecules. In one
aspect, extracellular
domains are receptors. Factors that bind the receptor domain include
circulating ligands, which may
be peptides, proteins, or small molecules such as adenosine and the like. For
example, growth
factors such as EGF, FGF and PDGF are circulating growth factors that bind to
their cognate
3 0 receptors to initiate a variety of cellular responses. Other factors
include cytokines, mitogenic factors,
neurotrophic factors and the like. Extracellular domains also bind to cell-
associated molecules. In this
respect, they mediate cell-cell interactions. Cell-associated ligands can be
tethered to the cell for
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example via a glycosylphosphatidylinositol (GPI) anchor, or may themselves be
transmembrane
proteins. Extracellular domains also associate with the extracellular matrix
and contribute to the
maintenance of the cell structure.
Putative transmembrane angiogenesis proteins include those encoded by the
sequences labeled with
"Y" in the TM column depicted in Table 2.
Angiogenesis proteins that are transmembrane are particularly preferred in the
present invention as
they are good targets for immunotherapeutics, as are described herein. In
addition, as outlined below,
transmembrane proteins can be also useful in imaging modalities.
It will also be appreciated by those in the art that a transmembrane protein
can be made soluble by
removing transmembrane sequences, for example through recombinant methods.
Furthermore,
transmembrane proteins that have been made soluble can be made to be secreted
through
recombinant means by adding an appropriate signal sequence.
In a preferred embodiment, the angiogenesis proteins are secreted proteins;
the secretion of which
can be either constitutive or regulated. These proteins have a signal peptide
or signal sequence that
targets the molecule to the secretory pathway. Secreted proteins are involved
in numerous
physiological events; by virtue of their circulating nature, they serve to
transmit signals to various other
cell types. The secreted protein may function in an autocrine manner (acting
on the cell that secreted
the factor), a paracrine manner (acting on cells in close proximity to the
cell that secreted the factor) or
an endocrine manner (acting on cells at a distance). Thus secreted molecules
find use in modulating
2 0 or altering numerous aspects of physiology. Angiogenesis proteins that are
secreted proteins are
particularly preferred in the present invention as they serve as good targets
for diagnostic markers, for
example for blood tests.
Putative secreted angiogenesis proteins include those encoded by the sequences
depicted in Table 2
that are labeled with "Y" in the SS column, but a "N" in the TM column.
2 5 An angiogenesis sequence is initially identified by substantial nucleic
acid and/or amino acid sequence
homology to the angiogenesis sequences outlined herein. Such homology can be
based upon the
overall nucleic acid or amino acid sequence, and is generally determined as
outlined below, using
either homology programs or hybridization conditions.
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As used herein, a nucleic acid is an "angiogenesis nucleic acid" if the
overall homology of the nucleic
acid sequence to one of the nucleic acids of Table 1, Table 2, Table 3, Table
4 or Table 5 is preferably
greater than about 75%, more preferably greater than about 80%, even more
preferably greater than
about 85% and most preferably greater than 90%. In some embodiments the
homology will be as
high as about 93 to 95 or 98%. Homology in this context means sequence
similarity or identity, with
identity being preferred. A preferred comparison for homology purposes is to
compare the sequence
containing sequencing errors to the correct sequence. This homology will be
determined using
standard techniques known in the art, including, but not limited to, the local
homology algorithm of
Smith & Waterman, Adv. Appl. Math. 2:482 (1981 ), by the homology alignment
algorith of Needleman
& Wunsch, J. Mol. Biool. 48:443 (1970), by the search for similarity method of
Pearson & Lipman,
PNAS USA 85:2444 (1988), by computerized implementations of these algorithms
(GAP, BESTFIT,
FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics
Computer Group, 575
Science Drive, Madison, WI), the Best Fit sequence program described by
Devereux et al., Nucl. Acid
Res. 12:387-395 (1984), preferably using the default settings, or by
inspection.
In a preferred embodiment, the sequences which are used to determine sequence
identity or similarity
are selected from the sequences set forth in the tables and figures,
preferable those represented in
Table 4, more preferably those represented in table 5, still more preferably
those of Figures 2, 3, 7, 11,
12, 17, 21, 23 and fragments thereof. In one embodiment the sequences utilized
herein are those set
forth in the tables and figures. In another embodiment, the sequences are
naturally occurring allelic
2 0 variants of the sequences set forth in the tables and figures. In another
embodiment, the sequences
are sequence variants as further described herein.
One example of a useful algorithm is PILEUP. PILEUP creates a multiple
sequence alignment from a
group of related sequences using progressive, pairwise alignments. It can also
plot a tree showing the
clustering relationships used to create the alignment. PILEUP uses a
simplification of the progressive
alignment method of Feng & Doolittle, J. Mol. Evol. 35:351-360 (1987); the
method is similar to that
described by Higgins & Sharp CABIOS 5:151-153 (1989). Useful PILEUP parameters
including a
default gap weight of 3.00, a default gap length weight of 0.10, and weighted
end gaps.
Another example of a useful algorithm is the BLAST algorithm, described in
Altschul et al., J. Mol. Biol.
215, 403-410, (1990) and Karlin et al., PNAS USA 90:5873-5787 (1993). A
particularly useful BLAST
3 0 program is the WU-BLAST-2 program which was obtained from Altschul et al.,
Methods in
Enzymology, 266: 460-480 (1996); http://blast.wustll. WU-BLAST-2 uses several
search parameters,
most of which are set to the default values. The adjustable parameters are set
with the following
values: overlap span =1, overlap fraction = 0.125, word threshold (T) = 11.
The HSP S and HSP S2
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parameters are dynamic values and are established by the program itself
depending upon the
composition of the particular sequence and composition of the particular
database against which the
sequence of interest is being searched; however, the values may be adjusted to
increase sensitivity.
A % amino acid sequence identity value is determined by the number of matching
identical residues
divided by the total number of residues of the "longer" sequence in the
aligned region. The "longer"
sequence is the one having the most actual residues in the aligned region
(gaps introduced by WU-
Blast-2 to maximize the alignment score are ignored).
Thus, "percent (%) nucleic acid sequence identity" is defined as the
percentage of nucleotide residues
in a candidate sequence that are identical with the nucleotide residues of the
nucleic acids of the
figures. A preferred method utilizes the BLASTN module of WU-BLAST-2 set to
the default
parameters, with overlap span and overlap fraction set to 1 and 0.125,
respectively.
The alignment may include the introduction of gaps in the sequences to be
aligned. In addition, for
sequences which contain either more or fewer nucleotides than those of the
nucleic acids of the
figures, it is understood that the percentage of homology will be determined
based on the number of
homologous nucleosides in relation to the total number of nucleosides. Thus,
for example, homology
of sequences shorter than those of the sequences identified herein and as
discussed below, will be
determined using the number of nucleosides in the shorter sequence.
In one embodiment, the nucleic acid homology is determined through
hybridization studies. Thus, for
example, nucleic acids which hybridize under high stringency to the nucleic
acids identified in the
2 0 figures, or their complements, are considered an angiogenesis sequence.
High stringency conditions
are known in the art; see for example Maniatis et al., Molecular Cloning: A
Laboratory Manual, 2d
Edition, 1989, and Short Protocols in Molecular Biology, ed. Ausubel, et al.,
both of which are hereby
incorporated by reference. Stringent conditions are sequence-dependent and
will be different in
different circumstances. Longer sequences hybridize specifically at higher
temperatures. An
extensive guide to the hybridization of nucleic acids is found in Tijssen,
Techniques in Biochemistry
and Molecular Biology--Hybridization with Nucleic Acid Probes, "Overview of
principles of hybridization
and the strategy of nucleic acid assays" (1993). Generally, stringent
conditions are selected to be
about 5-10°C lower than the thermal melting point (Tm) for the specific
sequence at a defined ionic
strength pH. The Tm is the temperature (under defined ionic strength, pH and
nucleic acid
3 0 concentration) at which 50% of the probes complementary to the target
hybridize to the target
sequence at equilibrium (as the target sequences are present in excess, at Tm,
50% of the probes are
occupied at equilibrium). Stringent conditions will be those in which the salt
concentration is less than
about 1.0 M sodium ion, typically about 0.01 to 1.0 M sodium ion concentration
(or other salts) at pH
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7.0 to 8.3 and the temperature is at least about 30°C for short probes
(e.g. 10 to 50 nucleotides) and
at least about 60°C for long probes (e.g. greater than 50 nucleotides).
Stringent conditions may also
be achieved with the addition of destabilizing agents such as formamide.
In another embodiment, less stringent hybridization conditions are used; for
example, moderate or low
stringency conditions may be used, as are known in the art; see Maniatis and
Ausubel, supra, and
Tijssen, supra.
In addition, the angiogenesis nucleic acid sequences of the invention are
fragments of larger genes,
i.e. they are nucleic acid segments. "Genes" in this context includes coding
regions, non-coding
regions, and mixtures of coding and non-coding regions. Accordingly, as will
be appreciated by those
in the art, using the sequences provided herein, additional sequences of the
angiogenesis genes can
be obtained, using techniques well known in the art for cloning either longer
sequences or the full
length sequences; see Maniatis et al., and Ausubel, et al., supra, hereby
expressly incorporated by
reference.
Once the angiogenesis nucleic acid is identified, it can be cloned and, if
necessary, its constituent
parts recombined to form the entire angiogenesis nucleic acid. Once isolated
from its natural source,
e.g., contained within a plasmid or other vector or excised therefrom as a
linear nucleic acid segment,
the recombinant angiogenesis nucleic acid can be further-used as a probe to
identify and isolate other
angiogenesis nucleic acids, for example additional coding regions. It can also
be used as a
"precursor" nucleic acid to make modified or variant angiogenesis nucleic
acids and proteins.
2 0 The angiogenesis nucleic acids of the present invention are used in
several ways. In a first
embodiment, nucleic acid probes to the angiogenesis nucleic acids are made and
attached to biochips
to be used in screening and diagnostic methods, as outlined below, or for
administration, for example
for gene therapy and/or antisense applications. Alternatively, the
angiogenesis nucleic acids that
include coding regions of angiogenesis proteins can be put into expression
vectors for the expression
2 5 of angiogenesis proteins, again either for screening purposes or for
administration to a patient.
In a preferred embodiment, nucleic acid probes to angiogenesis nucleic acids
(both the nucleic acid
sequences outlined in the figures and/or the complements thereof) are made.
The nucleic acid
probes attached to the biochip are designed to be substantially complementary
to the angiogenesis
nucleic acids, i.e. the target sequence (either the target sequence of the
sample or to other probe
3 0 sequences, for example in sandwich assays), such that hybridization of the
target sequence and the
probes of the present invention occurs. As outlined below, this
complementarity need not be perfect;
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there may be any number of base pair mismatches which will interfere with
hybridization between the
target sequence and the single stranded nucleic acids of the present
invention. However, if the
number of mutations is so great that no hybridization can occur under even the
least stringent of
hybridization conditions, the sequence is not a complementary target sequence.
Thus, by
"substantially complementary" herein is meant that the probes are sufficiently
complementary to the
target sequences to hybridize under normal reaction conditions, particularly
high stringency conditions,
as outlined herein.
A nucleic acid probe is generally single stranded but can be partially single
and partially double
stranded. The strandedness of the probe is dictated by the structure,
composition, and properties of
the target sequence. In general, the nucleic acid probes range from about 8 to
about 100 bases long,
with from about 10 to about 80 bases being preferred, and from about 30 to
about 50 bases being
particularly preferred. That is, generally whole genes are not used. In some
embodiments, much
longer nucleic acids can be used, up to hundreds of bases.
In a preferred embodiment, more than one probe per sequence is used, with
either overlapping
probes or probes to different sections of the target being used. That is, two,
three, four or more
probes, with three being preferred, are used to build in a redundancy for a
particular target. The
probes can be overlapping (i.e. have some sequence in common), or separate.
As will be appreciated by those in the art, nucleic acids can be attached or
immobilized to a solid
support in a wide variety of ways. By "immobilized" and grammatical
equivalents herein is meant the
2 0 association or binding between the nucleic acid probe and the solid
support is sufficient to be stable
under the conditions of binding, washing, analysis, and removal as outlined
below. The binding can be
covalent or non-covalent. By "non-covalent binding" and grammatical
equivalents herein is meant one
or more of either electrostatic, hydrophilic, and hydrophobic interactions.
Included in non-covalent
binding is the covalent attachment of a molecule, such as, streptavidin to the
support and the non-
2 5 covalent binding of the biotinylated probe to the streptavidin. By
"covalent binding" and grammatical
equivalents herein is meant that the two moieties, the solid support and the
probe, are attached by at
least one bond, including sigma bonds, pi bonds and coordination bonds.
Covalent bonds can be
formed directly between the probe and the solid support or can be formed by a
cross linker or by
inclusion of a specific reactive group on either the solid support or the
probe or both molecules.
3 0 Immobilization may also involve a combination of covalent and non-covalent
interactions.
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In general, the probes are attached to the biochip in a wide variety of ways,
as will be appreciated by
those in the art. As described herein, the nucleic acids can either be
synthesized first, with
subsequent attachment to the biochip, or can be directly synthesized on the
biochip.
The biochip comprises a suitable solid substrate. By "substrate" or "solid
support" or other
grammatical equivalents herein is meant any material that can be modified to
contain discrete
individual sites appropriate for the attachment or association of the nucleic
acid probes and is
amenable to at least one detection method. As will be appreciated by those in
the art, the number of
possible substrates are very large, and include, but are not limited to, glass
and modified or
functionalized glass, plastics (including acrylics, polystyrene and copolymers
of styrene and other
materials, polypropylene, polyethylene, polybutylene, polyurethanes, TefIonJ,
etc.), polysaccharides,
nylon or nitrocellulose, resins, silica or silica-based materials including
silicon and modified silicon,
carbon, metals, inorganic glasses, plastics, etc. In general, the substrates
allow optical detection and
do not appreciably fluorescese. A preferred substrate is described in
copending application entitled
Reusable Low Fluorescent Plastic Biochip, U.S. Application Serial No.
09/270,214, filed March 15,
1999, herein incorporated by reference in its entirety.
Generally the substrate is planar, although as will be appreciated by those in
the art, other
configurations of substrates may be used as well. For example, the probes may
be placed on the
inside surface of a tube, for flow-through sample analysis to minimize sample
volume. Similarly, the
substrate may be flexible, such as a flexible foam, including closed cell
foams made of particular
2 0 plastics.
In a preferred embodiment, the surface of the biochip and the probe may be
derivatized with chemical
functional groups for subsequent attachment of the two. Thus, for example, the
biochip is derivatized
with a chemical functional group including, but not limited to, amino groups,
carboxy groups, oxo
groups and thiol groups, with amino groups being particularly preferred. Using
these functional
2 5 groups, the probes can be attached using functional groups on the probes.
For example, nucleic
acids containing amino groups can be attached to surfaces comprising amino
groups, for example
using linkers as are known in the art; for example, homo-or hetero-
bifunctional linkers as are well
known (see 1994 Pierce Chemical Company catalog, technical section on cross-
linkers, pages
155-200, incorporated herein by reference). In addition, in some cases,
additional linkers, such as
3 0 alkyl groups (including substituted and heteroalkyl groups) may be used.
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In this embodiment, the oligonucleotides are synthesized as is known in the
art, and then attached to
the surface of the solid support. As will be appreciated by those skilled in
the art, either the 5' or 3'
terminus may be attached to the solid support, or attachment may be via an
internal nucleoside.
In an additional embodiment, the immobilization to the solid support may be
very strong, yet non-
covalent. For example, biotinylated oligonucleotides can be made, which bind
to surfaces covalently
coated with streptavidin, resulting in attachment.
Alternatively, the oligonucleotides may be synthesized on the surface, as is
known in the art. For
example, photoactivation techniques utilizing photopolymerization compounds
and techniques are
used. In a preferred embodiment, the nucleic acids can be synthesized in situ,
using well known
photolithographic techniques, such as those described in WO 95/25116; WO
95/35505; U.S. Patent
Nos. 5,700,637 and 5,445,934; and references cited within, all of which are
expressly incorporated by
reference; these methods of attachment form the basis of the Affimetrix
GeneChipT"' technology.
In a preferred embodiment, angiogenesis nucleic acids encoding angiogenesis
proteins are used to
make a variety of expression vectors to express angiogenesis proteins which
can then be used in
screening assays, as described below. The expression vectors may be either
self-replicating
extrachromosomal vectors or vectors which integrate into a host genome.
Generally, these
expression vectors include transcriptional and translational regulatory
nucleic acid operably linked to
the nucleic acid encoding the angiogenesis protein. The term "control
sequences" refers to DNA
sequences necessary for the expression of an operably linked coding sequence
in a particular host
2 0 organism. The control sequences that are suitable for prokaryotes, for
example, include a promoter,
optionally an operator sequence, and a ribosome binding site. Eukaryotic cells
are known to utilize
promoters, polyadenylation signals, and enhancers.
Nucleic acid is "operably linked" when it is placed into a functional
relationship with another nucleic
2 5 acid sequence. For example, DNA for a presequence or secretory leader is
operably linked to DNA
for a polypeptide if it is expressed as a preprotein that participates in the
secretion of the polypeptide;
a promoter or enhancer is operably linked to a coding sequence if it affects
the transcription of the
sequence; or a ribosome binding site is operably linked to a coding sequence
if it is positioned so as to
facilitate translation. Generally, "operably linked" means that the DNA
sequences being linked are
3 0 contiguous, and, in the case of a secretory leader, contiguous and in
reading phase. However,
enhancers do not have to be contiguous. Linking is accomplished by ligation at
convenient restriction
sites. If such sites do not exist, the synthetic oligonucleotide adaptors or
linkers are used in
accordance with conventional practice. The transcriptional and translational
regulatory nucleic acid
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will generally be appropriate to the host cell used to express the
angiogenesis protein; for example,
transcriptional and translational regulatory nucleic acid sequences from
Bacillus are preferably used to
express the angiogenesis protein in Bacillus. Numerous types of appropriate
expression vectors, and
suitable regulatory sequences are known in the art for a variety of host
cells.
In general, the transcriptional and translational regulatory sequences may
include, but are not limited
to, promoter sequences, ribosomal binding sites, transcriptional start and
stop sequences,
translational start and stop sequences, and enhancer or activator sequences.
In a preferred
embodiment, the regulatory sequences include a promoter and transcriptional
start and stop
sequences.
Promoter sequences encode either constitutive or inducible promoters. The
promoters may be either
naturally occurring promoters or hybrid promoters. Hybrid promoters, which
combine elements of
more than one promoter, are also known in the art, and are useful in the
present invention.
In addition, the expression vector may comprise additional elements. For
example, the expression
vector may have two replication systems, thus allowing it to be maintained in
two organisms, for
example in mammalian or insect cells for expression and in a procaryotic host
for cloning and
amplification. Furthermore, for integrating expression vectors, the expression
vector contains at least
one sequence homologous to the host cell genome, and preferably two homologous
sequences which
flank the expression construct. The integrating vector may be directed to a
specific locus in the host
cell by selecting the appropriate homologous sequence for inclusion in the
vector. Constructs for
2 0 integrating vectors are well known in the art.
In addition, in a preferred embodiment, the expression vector contains a
selectable marker gene to
allow the selection of transformed host cells. Selection genes are well known
in the art and will vary
with the host cell used.
The angiogenesis proteins of the present invention are produced by culturing a
host cell transformed
2 5 with an expression vector containing nucleic acid encoding an angiogenesis
protein, under the
appropriate conditions to induce or cause expression of the angiogenesis
protein. The conditions
appropriate for angiogenesis protein expression will vary with the choice of
the expression vector and
the host cell, and will be easily ascertained by one skilled in the art
through routine experimentation.
For example, the use of constitutive promoters in the expression vector will
require optimizing the
3 0 growth and proliferation of the host cell, while the use of an inducible
promoter requires the
appropriate growth conditions for induction. In addition, in some embodiments,
the timing of the
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WO 01/11086 PCT/US00/22061
harvest is important. For example, the baculoviral systems used in insect cell
expression are lytic
viruses, and thus harvest time selection can be crucial for product yield.
Appropriate host cells include yeast, bacteria, archaebacteria, fungi, and
insect and animal cells,
including mammalian cells. Of particular interest are Drosophila melangaster
cells, Saccharomyces
cerevisiae and other yeasts, E. coli, Bacillus subtilis, Sf9 cells, C129
cells, 293 cells, Neurospora,
BHK, CHO, COS, HeLa cells, HUVEC (human umbilical vein endothelial cells),THP1
cells (a
macrophage cell line) and human cells and lines.
In a preferred embodiment, the angiogenesis proteins are expressed in
mammalian cells. Mammalian
expression systems are also known in the art, and include retroviral systems.
A preferred expression
vector system is a retroviral vector system such as is generally described in
PCT/US97/01019 and
PCT/US97/01048, both of which are hereby expressly incorporated by reference.
Of particular use as
mammalian promoters are the promoters from mammalian viral genes, since the
viral genes are often
highly expressed and have a broad host range. Examples include the SV40 early
promoter, mouse
mammary tumor virus LTR promoter, adenovirus major late promoter, herpes
simplex virus promoter,
and the CMV promoter. Typically, transcription termination and polyadenylation
sequences
recognized by mammalian cells are regulatory regions located 3' to the
translation stop codon and
thus, together with the promoter elements, flank the coding sequence. Examples
of transcription
terminator and polyadenlytion signals include those derived form SV40.
The methods of introducing exogenous nucleic acid into mammalian hosts, as
well as other hosts, is
2 0 well known in the art, and will vary with the host cell used. Techniques
include dextrin-mediated
transfection, calcium phosphate precipitation, polybrene mediated
transfection, protoplast fusion,
electroporation, viral infection, encapsulation of the polynucleotide(s) in
liposomes, and direct
microinjection of the DNA into nuclei.
In a preferred embodiment, angiogenesis proteins are expressed in bacterial
systems. Bacterial
2 5 expression systems are well known in the art. Promoters from bacteriophage
may also be used and
are known in the art. In addition, synthetic promoters and hybrid promoters
are also useful; for
example, the tic promoter is a hybrid of the trp and lac promoter sequences.
Furthermore, a bacterial
promoter can include naturally occurring promoters of non-bacterial origin
that have the ability to bind
bacterial RNA polymerise and initiate transcription. In addition to a
functioning promoter sequence,
3 0 an efficient ribosome binding site is desirable. The expression vector may
also include a signal
peptide sequence that provides for secretion of the angiogenesis protein in
bacteria. The protein is
either secreted into the growth media (gram-positive bacteria) or into the
periplasmic space, located
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between the inner and outer membrane of the cell (gram-negative bacteria). The
bacterial expression
vector may also include a selectable marker gene to allow for the selection of
bacterial strains that
have been transformed. Suitable selection genes include genes which render the
bacteria resistant to
drugs such as ampicillin, chloramphenicol, erythromycin, kanamycin, neomycin
and tetracycline.
Selectable markers also include biosynthetic genes, such as those in the
histidine, tryptophan and
leucine biosynthetic pathways. These components are assembled into expression
vectors.
Expression vectors for bacteria are well known in the art, and include vectors
for Bacillus subtilis, E.
coli, Streptococcus cremoris, and Streptococcus lividans, among others. The
bacterial expression
vectors are transformed into bacterial host cells using techniques well known
in the art, such as
calcium chloride treatment, electroporation, and others.
In one embodiment, angiogenesis proteins are produced in insect cells.
Expression vectors for the
transformation of insect cells, and in particular, baculovirus-based
expression vectors, are well known
in the art.
In a preferred embodiment, angiogenesis protein is produced in yeast cells.
Yeast expression
systems are well known in the art, and include expression vectors for
Saccharomyces cerevisiae,
Candida albicans and C. maltosa, Hansenula polymorpha, Kluyveromyces fragilis
and K. lactis, Pichia
guillerimondii and P. pasforis, Schizosaccharomyces pombe, and Yarrowia
lipolytica.
The angiogenesis protein may also be made as a fusion protein, using
techniques well known in the
art. Thus, for example, for the creation of monoclonal antibodies, if the
desired epitope is small, the
2 0 angiogenesis protein may be fused to a carrier protein to form an
immunogen. Alternatively, the
angiogenesis protein may be made as a fusion protein to increase expression,
or for other reasons.
For example, when the angiogenesis protein is an angiogenesis peptide, the
nucleic acid encoding the
peptide may be linked to other nucleic acid for expression purposes.
In one embodiment, the angiogenesis nucleic acids, proteins and antibodies of
the invention are
2 5 labeled. By "labeled" herein is meant that a compound has at least one
element, isotope or chemical
compound attached to enable the detection of the compound. In general, labels
fall into three classes:
a) isotopic labels, which may be radioactive or heavy isotopes; b) immune
labels, which may be
antibodies or antigens; and c) colored or fluorescent dyes. The labels may be
incorporated into the
angiogenesis nucleic acids, proteins and antibodies at any position. For
example, the label should be
3 0 capable of producing, either directly or indirectly, a detectable signal.
The detectable moiety may be a
radioisotope, such as 3H, "C, 32P, ssS, or '251, a fluorescent or
chemiluminescent compound, such as
fluorescein isothiocyanate, rhodamine, or luciferin, or an enzyme, such as
alkaline phosphatase, beta
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galactosidase or horseradish peroxidase. Any method known in the art for
conjugating the antibody to
the label may be employed, including those methods described by Hunter et al.,
Nature, 144:945
(1962); David et al., Biochemistry, 13:1014 (1974); Pain et al., J. Immunol.
Meth., 40:219 (1981); and
Nygren, J. Histochem. and Cytochem., 30:407 (1982).
Accordingly, the present invention also provides angiogenesis protein
sequences. An angiogenesis
protein of the present invention may be identified in several ways. "Protein"
in this sense includes
proteins, polypeptides, and peptides. As will be appreciated by those in the
art, the nucleic acid
sequences of the invention can be used to generate protein sequences. There
are a variety of ways
to do this, including cloning the entire gene and verifying its frame and
amino acid sequence, or by
comparing it to known sequences to search for homology to provide a frame,
assuming the
angiogenesis protein has homology to some protein in the database being used.
Generally, the
nucleic acid sequences are input into a program that will search all three
frames for homology. This is
done in a preferred embodiment using the following NCBI Advanced BLAST
parameters. The program
is blastx or blastn. The database is nr. The input data is as "Sequence in
FASTA format". The
organism list is "none". The "expect" is 10; the filter is default. The
"descriptions" is 500, the
"alignments" is 500, and the "alignment view" is pairwise. The "Query Genetic
Codes" is standard (1 ).
The matrix is BLOSUM62; gap existence cost is 11, per residue gap cost is 1;
and the lambda ratio is
85 default. This results in the generation of a putative protein sequence.
Also included within one embodiment of angiogenesis proteins are amino acid
variants of the naturally
2 0 occurring sequences, as determined herein. Preferably, the variants are
preferably greater than about
75% homologous to the wild-type sequence, more preferably greater than about
80%, even more
preferably greater than about 85% and most preferably greater than 90%. In
some embodiments the
homology will be as high as about 93 to 95 or 98%. As for nucleic acids,
homology in this context
means sequence similarity or identity, with identity being preferred. This
homology will be determined
2 5 using standard techniques known in the art as are outlined above for the
nucleic acid homologies.
Angiogenesis proteins of the present invention may be shorter or longer than
the wild type amino acid
sequences. Thus, in a preferred embodiment, included within the definition of
angiogenesis proteins
are portions or fragments of the wild type sequences. herein. In addition, as
outlined above, the
angiogenesis nucleic acids of the invention may be used to obtain additional
coding regions, and thus
3 0 additional protein sequence, using techniques known in the art.
In a preferred embodiment, the angiogenesis proteins are derivative or variant
angiogenesis proteins
as compared to the wild-type sequence. That is, as outlined more fully below,
the derivative
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angiogenesis peptide will contain at least one amino acid substitution,
deletion or insertion, with amino
acid substitutions being particularly preferred. The amino acid substitution,
insertion or deletion may
occur at any residue within the angiogenesis peptide.
Also included within one embodiment of angiogenesis proteins of the present
invention are amino acid
sequence variants. These variants fall into one or more of three classes:
substitutional, insertional or
deletional variants. These variants ordinarily are prepared by site specific
mutagenesis of nucleotides
in the DNA encoding the angiogenesis protein, using cassette or PCR
mutagenesis or other
techniques well known in the art, to produce DNA encoding the variant, and
thereafter expressing the
DNA in recombinant cell culture as outlined above. However, variant
angiogenesis protein fragments
having up to about 100-150 residues may be prepared by in vitro synthesis
using established
techniques. Amino acid sequence variants are characterized by the
predetermined nature of the
variation, a feature that sets them apart from naturally occurring allelic or
interspecies variation of the
angiogenesis protein amino acid sequence. The variants typically exhibit the
same qualitative
biological activity as the naturally occurring analogue, although variants can
also be selected which
have modified characteristics as will be more fully outlined below.
While the site or region for introducing an amino acid sequence variation is
predetermined, the
mutation per se need not be predetermined. For example, in order to optimize
the performance of a
mutation at a given site, random mutagenesis may be conducted at the target
codon or region and the
expressed angiogenesis variants screened for the optimal combination of
desired activity. Techniques
2 0 for making substitution mutations at predetermined sites in DNA having a
known sequence are well
known, for example, M13 primer mutagenesis and PCR mutagenesis. Screening of
the mutants is
done using assays of angiogenesis protein activities.
Amino acid substitutions are typically of single residues; insertions usually
will be on the order of from
about 1 to 20 amino acids, although considerably larger insertions may be
tolerated. Deletions range
2 5 from about 1 to about 20 residues, although in some cases deletions may be
much larger.
Substitutions, deletions, insertions or any combination thereof may be used to
arrive at a final
derivative. Generally these changes are done on a few amino acids to minimize
the alteration of the
molecule. However, larger changes may be tolerated in certain circumstances.
When small
alterations in the characteristics of the angiogenesis protein are desired,
substitutions are generally
3 0 made in accordance with the following chart:
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Chart I
Original Residue Exemplary Substitutions
Ala Ser
Arg Lys
Asn Gln, His
Asp Glu
Cys Ser
Gln Asn
Glu Asp
Gly Pro
His Asn, Gln
Ile Leu, Val
Leu Ile, Val
Lys Arg, Gln, Glu
Met Leu, Ile
Phe Met, Leu, Tyr
Ser Thr
Thr Ser
Trp Tyr
2 0 Tyr Trp, Phe
Val Ile, Leu
Substantial changes in function or immunological identity are made by
selecting substitutions that are
less conservative than those shown in Chart I. For example, substitutions may
be made which more
significantly affect: the structure of the polypeptide backbone in the area of
the alteration, for example
2 5 the alpha-helical or beta-sheet structure; the charge or hydrophobicity of
the molecule at the target
site; or the bulk of the side chain. The substitutions which in general are
expected to produce the
greatest changes in the polypeptide's properties are those in which (a) a
hydrophilic residue, e.g. seryl
or threonyl, is substituted for (or by) a hydrophobic residue, e.g. leucyl,
isoleucyl, phenylalanyl, valyl or
alanyl; (b) a cysteine or proline is substituted for (or by) any other
residue; (c) a residue having an
3 0 electropositive side chain, e.g. lysyl, arginyl, or histidyl, is
substituted for (or by) an electronegative
residue, e.g. glutamyl or aspartyl; or (d) a residue having a bulky side
chain, e.g. phenylalanine, is
substituted for (or by) one not having a side chain, e.g. glycine.
The variants typically exhibit the same qualitative biological activity and
will elicit the same immune
response as the naturally-occurring analogue, although variants also are
selected to modify the
3 5 characteristics of the angiogenesis proteins as needed. Alternatively, the
variant may be designed
such that the biological activity of the angiogenesis protein is altered. For
example, glycosylation sites
may be altered or removed.
Covalent modifications of angiogenesis polypeptides are included within the
scope of this invention.
One type of covalent modification includes reacting targeted amino acid
residues of an angiogenesis
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polypeptide with an organic derivatizing agent that is capable of reacting
with selected side chains or
the N-or C-terminal residues of an angiogenesis polypeptide. Derivatization
with bifunctional agents is
useful, for instance, for crosslinking angiogenesis polypeptides to a water-
insoluble support matrix or
surface for use in the method for purifying anti-angiogenesis polypeptide
antibodies or screening
assays, as is more fully described below. Commonly used crosslinking agents
include, e.g., 1,1-
bis(diazoacetyl)-2-phenylethane, glutaraldehyde, N-hydroxysuccinimide esters,
for example, esters
with 4-azidosalicylic acid, homobifunctional imidoesters, including
disuccinimidyl esters such as 3,3'-
dithiobis(succinimidylpropionate), bifunctional maleimides such as bis-N-
maleimido-1,8-octane and
agents such as methyl-3-[(p-azidophenyl)dithio]propioimidate.
Other modifications include deamidation of glutaminyl and asparaginyl residues
to the corresponding
glutamyl and aspartyl residues, respectively, hydroxylation of proline and
lysine, phosphorylation of
hydroxyl groups of Beryl, threonyl or tyrosyl residues, methylation of the a-
amino groups of lysine,
arginine, and histidine side chains [T.E. Creighton, Proteins: Structure and
Molecular Properties, W.H.
Freeman & Co., San Francisco, pp. 79-86 (1983)], acetylation of the N-terminal
amine, and amidation
of any C-terminal carboxyl group.
Another type of covalent modification of the angiogenesis polypeptide included
within the scope of this
invention comprises altering the native glycosylation pattern of the
polypeptide. "Altering the native
2 0 glycosylation pattern" is intended for purposes herein to mean deleting
one or more carbohydrate
moieties found in native sequence angiogenesis polypeptide, and/or adding one
or more glycosylation
sites that are not present in the native sequence angiogenesis polypeptide.
Addition of glycosylation sites to angiogenesis polypeptides may be
accomplished by altering the
2 5 amino acid sequence thereof. The alteration may be made, for example, by
the addition of, or
substitution by, one or more serine or threonine residues to the native
sequence angiogenesis
polypeptide (for O-linked glycosylation sites). The angiogenesis amino acid
sequence may optionally
be altered through changes at the DNA level, particularly by mutating the DNA
encoding the
angiogenesis polypeptide at preselected bases such that codons are generated
that will translate into
3 0 the desired amino acids.
Another means of increasing the number of carbohydrate moieties on the
angiogenesis polypeptide is
by chemical or enzymatic coupling of glycosides to the polypeptide. Such
methods are described in
the art, e.g., in WO 87/05330 published 11 September 1987, and in Aplin and
Wriston, CRC Crit. Rev.
Biochem., pp. 259-306 (1981).
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Removal of carbohydrate moieties present on the angiogenesis polypeptide may
be accomplished
chemically or enzymatically or by mutational substitution of codons encoding
for amino acid residues
that serve as targets for glycosylation. Chemical deglycosylation techniques
are known in the art and
described, for instance, by Hakimuddin, et al., Arch. Biochem. Biophys.,
259:52 (1987) and by Edge et
al., Anal. Biochem., 118:131 (1981 ). Enzymatic cleavage of carbohydrate
moieties on polypeptides
can be achieved by the use of a variety of endo-and exo-glycosidases as
described by Thotakura et
al., Meth. Enzymol., 138:350 (1987).
Another type of covalent modification of angiogenesis comprises linking the
angiogenesis polypeptide
to one of a variety of nonproteinaceous polymers, e.g., polyethylene glycol,
polypropylene glycol, or
polyoxyalkylenes, in the manner set forth in U.S. Patent Nos. 4,640,835;
4,496,689; 4,301,144;
4,670,417; 4,791,192 or 4,179,337.
Angiogenesis polypeptides of the present invention may also be modified in a
way to form chimeric
molecules comprising an angiogenesis polypeptide fused to another,
heterologous polypeptide or
amino acid sequence. In one embodiment, such a chimeric molecule comprises a
fusion of an
angiogenesis polypeptide with a tag polypeptide which provides an epitope to
which an anti-tag
antibody can selectively bind. The epitope tag is generally placed at the
amino-or carboxyl-terminus of
the angiogenesis polypeptide. The presence of such epitope-tagged forms of an
angiogenesis
polypeptide can be detected using an antibody against the tag polypeptide.
Also, provision of the
epitope tag enables the angiogenesis polypeptide to be readily purified by
affinity purification using an
2 0 anti-tag antibody or another type of affinity matrix that binds to the
epitope tag. In an alternative
embodiment, the chimeric molecule may comprise a fusion of an angiogenesis
polypeptide with an
immunoglobulin or a particular region of an immunoglobulin. For a bivalent
form of the chimeric
molecule, such a fusion could be to the Fc region of an IgG molecule.
Various tag polypeptides and their respective antibodies are well known in the
art. Examples include
poly-histidine (poly-his) or poly-histidine-glycine (poly-his-gly) tags; the
flu HA tag polypeptide and its
antibody 12CA5 [Field et al., Mol. Cell. Biol., 8:2159-2165 (1988)]; the c-myc
tag and the 8F9, 3C7,
6E10, G4, B7 and 9E10 antibodies thereto [Evan et al., Molecular and Cellular
Biology, 5:3610-3616
(1985)]; and the Herpes Simplex virus glycoprotein D (gD) tag and its antibody
[Paborsky et al.,
Protein Engineering, 3(6):547-553 (1990)]. Other tag polypeptides include the
Flag-peptide [Hopp et
3 0 al., BioTechnology, 6:1204-1210 (1988)]; the KT3 epitope peptide [Martin
et al., Science, 255:192-194
(1992)]; tubulin epitope peptide [Skinner et al., J. Biol. Chem., 266:15163-
15166 (1991 )]; and the T7
gene 10 protein peptide tag [Lutz-Freyermuth et al., Proc. Natl. Acad. Sci.
USA, 87:6393-6**397
(1990)].
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Also included with an embodiment of angiogenesis protein are other
angiogenesis proteins of the
angiogenesis family, and angiogenesis proteins from other organisms, which are
cloned and
expressed as outlined below. Thus, probe or degenerate polymerase chain
reaction (PCR) primer
sequences may be used to find other related angiogenesis proteins from humans
or other organisms.
As will be appreciated by those in the art, particularly useful probe and/or
PCR primer sequences
include the unique areas of the angiogenesis nucleic acid sequence. As is
generally known in the art,
preferred PCR primers are from about 15 to about 35 nucleotides in length,
with from about 20 to
about 30 being preferred, and may contain inosine as needed. The conditions
for the PCR reaction
are well known in the art.
In addition, as is outlined herein, angiogenesis proteins can be made that are
longer than those
encoded by the nucleic acids of the figures, for example, by the elucidation
of additional sequences,
the addition of epitope or purification tags, the addition of other fusion
sequences, etc.
Angiogenesis proteins may also be identified as being encoded by angiogenesis
nucleic acids. Thus,
angiogenesis proteins are encoded by nucleic acids that will hybridize to the
sequences of the
sequence listings, or their complements, as outlined herein.
In a preferred embodiment, when the angiogenesis protein is to be used to
generate antibodies, for
example for immunotherapy, the angiogenesis protein should share at least one
epitope or
determinant with the full length protein. By "epitope" or "determinant" herein
is meant a portion of a
protein which will generate and/or bind an antibody or T-cell receptor in the
context of MHC. Thus, in
2 0 most instances, antibodies made to a smaller angiogenesis protein will be
able to bind to the full
length protein. In a preferred embodiment, the epitope is unique; that is,
antibodies generated to a
unique epitope show little or no cross-reactivity. In a preferred embodiment,
the epitope is selected
from AAA4p1 and AAA4p2. In another preferred embodiment the epitope is
selected from AAA1p1
and AAA1 p2. In another preferred embodiment the epitope is selected from
AAA7p1, AAA7p2,
2 5 AAA7p3 and AAA7p1 m.
In one embodiment, the term "antibody" includes antibody fragments, as are
known in the art,
including Fab, Fab2, single chain antibodies (Fv for example), chimeric
antibodies, etc., either
produced by the modification of whole antibodies or those synthesized de novo
using recombinant
DNA technologies.
3 0 Methods of preparing polyclonal antibodies are known to the skilled
artisan. Polyclonal antibodies can
be raised in a mammal, for example, by one or more injections of an immunizing
agent and, if desired,
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an adjuvant. Typically, the immunizing agent and/or adjuvant will be injected
in the mammal by
multiple subcutaneous or intraperitoneal injections. The immunizing agent may
include a protein
encoded by a nucleic acid of the figures or fragment thereof or a fusion
protein thereof. It may be
useful to conjugate the immunizing agent to a protein known to be immunogenic
in the mammal being
immunized. Examples of such immunogenic proteins include but are not limited
to keyhole limpet
hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin
inhibitor. Examples of
adjuvants which may be employed include Freund's complete adjuvant and MPL-TDM
adjuvant
(monophosphoryl Lipid A, synthetic trehalose dicorynomycolate). The
immunization protocol may be
selected by one skilled in the art without undue experimentation.
The antibodies may, alternatively, be monoclonal antibodies. Monoclonal
antibodies may be prepared
using hybridoma methods, such as those described by Kohler and Milstein,
Nature, 256:495 (1975).
In a hybridoma method, a mouse, hamster, or other appropriate host animal, is
typically immunized
with an immunizing agent to elicit lymphocytes that produce or are capable of
producing antibodies
that will specifically bind to the immunizing agent. Alternatively, the
lymphocytes may be immunized in
vitro. The immunizing agent will typically include a polypeptide encoded by a
nucleic acid of Table 1,
Table 2, Table 3, Table 4 or Table 5 or fragment thereof or a fusion protein
thereof. Generally, either
peripheral blood lymphocytes ("PBLs") are used if cells of human origin are
desired, or spleen cells or
lymph node cells are used if non-human mammalian sources are desired. The
lymphocytes are then
fused with an immortalized cell line using a suitable fusing agent, such as
polyethylene glycol, to form
2 0 a hybridoma cell [coding, Monoclonal Antibodies: Principles and Practice,
Academic Press, (1986) pp.
59-103]. Immortalized cell lines are usually transformed mammalian cells,
particularly myeloma cells
of rodent, bovine and human origin. Usually, rat or mouse myeloma cell lines
are employed. The
hybridoma cells may be cultured in a suitable culture medium that preferably
contains one or more
substances that inhibit the growth or survival of the unfused, immortalized
cells. For example, if the
parental cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase
(HGPRT or HPRT),
the culture medium for the hybridomas typically will include hypoxanthine,
aminopterin, and thymidine
("HAT medium"), which substances prevent the growth of HGPRT-deficient cells.
In one embodiment, the antibodies are bispecific antibodies. Bispecific
antibodies are monoclonal,
preferably human or humanized, antibodies that have binding specificities for
at least two different
3 0 antigens. In the present case, one of the binding specificities is for a
protein encoded by a nucleic
acid of figure 1 or 3-6 or a fragment thereof, the other one is for any other
antigen, and preferably for a
cell-surface protein or receptor or receptor subunit, preferably one that is
tumor specific.
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In a preferred embodiment, the antibodies to angiogenesis protein are capable
of reducing or
eliminating the biological function of angiogenesis protein, as is described
below. That is, the addition
of anti-angiogenesis protein antibodies (either polyclonal or preferably
monoclonal) to angiogenic
tissue (or cells containing angiogenesis) may reduce or eliminate the
angiogenesis activity. Generally,
at least a 25% decrease in activity is preferred, with at least about 50%
being particularly preferred
and about a 95-100% decrease being especially preferred.
In a preferred embodiment the antibodies to the angiogenesis proteins are
humanized antibodies.
Humanized forms of non-human (e.g., murine) antibodies are chimeric molecules
of immunoglobulins,
immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')2 or
other antigen-binding
subsequences of antibodies) which contain minimal sequence derived from non-
human
immunoglobulin. Humanized antibodies include human immunoglobulins (recipient
antibody) in which
residues form a complementary determining region (CDR) of the recipient are
replaced by residues
from a CDR of a non-human species (donor antibody) such as mouse, rat or
rabbit having the desired
specificity, affinity and capacity. In some instances, Fv framework residues
of the human
immunoglobulin are replaced by corresponding non-human residues. Humanized
antibodies may also
comprise residues which are found neither in the recipient antibody nor in the
imported CDR or
framework sequences. In general, the humanized antibody will comprise
substantially all of at least
one, and typically two, variable domains, in which all or substantially all of
the CDR regions correspond
to those of a non-human immunoglobulin and all or substantially all of the FR
regions are those of a
2 0 human immunoglobulin consensus sequence. The humanized antibody optimally
also will comprise at
least a portion of an immunoglobulin constant region (Fc), typically that of a
human immunoglobulin
[Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-
329 (1988); and Presta,
Curr. Op. Struct. Biol., 2:593-596 (1992)].
Methods for humanizing non-human antibodies are well known in the art.
Generally, a humanized
2 5 antibody has one or more amino acid residues introduced into it from a
source which is non-human.
These non-human amino acid residues are often referred to as import residues,
which are typically
taken from an import variable domain. Humanization can be essentially
performed following the
method of Winter and co-workers [Jones et al., Nature, 321:522-525 (1986);
Riechmann et al., Nature,
332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)], by
substituting rodent CDRs
3 0 or CDR sequences for the corresponding sequences of a human antibody.
Accordingly, such
humanized antibodies are chimeric antibodies (U.S. Patent No. 4,816,567),
wherein substantially less
than an intact human variable domain has been substituted by the corresponding
sequence from a
non-human species. In practice, humanized antibodies are typically human
antibodies in which some
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CDR residues and possibly some FR residues are substituted by residues from
analogous sites in
rodent antibodies.
Human antibodies can also be produced using various techniques known in the
art, including phage
display libraries (Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991 );
Marks et al., J. Mol. Biol.,
222:581 (1991 )]. The techniques of Cole et al. and Boerner et al. are also
available for the preparation
of human monoclonal antibodies (Cole et al., Monoclonal Antibodies and Cancer
Therapy, Alan R.
Liss, p. 77 (1985) and Boerner et al., J. Immunol., 147 1 :86-95 (1991 )].
Similarly, human antibodies
can be made by introducing of human immunoglobulin loci into transgenic
animals, e.g., mice in which
the endogenous immunoglobulin genes have been partially or completely
inactivated. Upon
challenge, human antibody production is observed, which closely resembles that
seen in humans in all
respects, including gene rearrangement, assembly, and antibody repertoire.
This approach is
described, for example, in U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825;
5,625,126; 5,633,425;
5,661,016, and in the following scientific publications: Marks et al.,
Bio/Technoloay 10, 779-783
(1992); Lonberg et al., Nature 368 856-859 (1994); Morrison, Nature 368, 812-
13 (1994); Fishwild et
al., Nature Biotechnolo4y 14, 845-51 (1996); Neuberger, Nature Biotechnology
14, 826 (1996);
Lonberg and Huszar, Intern. Rev. Immunol. 13 65-93 (1995).
By immunotherapy is meant treatment of angiogenesis with an antibody raised
against angiogenesis
proteins. As used herein, immunotherapy can be passive or active. Passive
immunotherapy as
defined herein is the passive transfer of antibody to a recipient (patient).
Active immunization is the
2 0 induction of antibody and/or T-cell responses in a recipient (patient).
Induction of an immune
response is the result of providing the recipient with an antigen to which
antibodies are raised. As
appreciated by one of ordinary skill in the art, the antigen may be provided
by injecting a polypeptide
against which antibodies are desired to be raised into a recipient, or
contacting the recipient with a
nucleic acid capable of expressing the antigen and under conditions for
expression of the antigen.
2 5 In a preferred embodiment the angiogenesis proteins against which
antibodies are raised are secreted
proteins as described above. Without being bound by theory, antibodies used
for treatment, bind and
prevent the secreted protein from binding to its receptor, thereby
inactivating the secreted
angiogenesis protein.
In another preferred embodiment, the angiogenesis protein to which antibodies
are raised is a
3 0 transmembrane protein. Without being bound by theory, antibodies used for
treatment, bind the
extracellular domain of the angiogenesis protein and prevent it from binding
to other proteins, such as
circulating ligands or cell-associated molecules. The antibody may cause down-
regulation of the
CA 02381699 2002-02-07
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transmembrane angiogenesis protein. As will be appreciated by one of ordinary
skill in the art, the
antibody may be a competitive, non-competitive or uncompetitive inhibitor of
protein binding to the
extracellular domain of the angiogenesis protein. The antibody is also an
antagonist of the
angiogenesis protein. Further, the antibody prevents activation of the
transmembrane angiogenesis
protein. In one aspect, when the antibody prevents the binding of other
molecules to the angiogenesis
protein, the antibody prevents growth of the cell. The antibody also
sensitizes the cell to cytotoxic
agents, including, but not limited to TNF-a, TNF-~, IL-1, INF-y and IL-2, or
chemotherapeutic agents
including SFU, vinblastine, actinomycin D, cisplatin, methotrexate, and the
like. In some instances the
antibody belongs to a sub-type that activates serum complement when complexed
with the
transmembrane protein thereby mediating cytotoxicity. Thus, angiogenesis is
treated by administering
to a patient antibodies directed against the transmembrane angiogenesis
protein.
In another preferred embodiment, the antibody is conjugated to a therapeutic
moiety. In one aspect
the therapeutic moiety is a small molecule that modulates the activity of the
angiogenesis protein. In
another aspect the therapeutic moiety modulates the activity of molecules
associated with or in close
proximity to the angiogenesis protein. The therapeutic moiety may inhibit
enzymatic activity such as
protease or collagenase activity associated with angiogenesis.
In a preferred embodiment, the therapeutic moiety may also be a cytotoxic
agent. In this method,
targeting the cytotoxic agent to angiogenesis tissue or cells, results in a
reduction in the number of
afflicted cells, thereby reducing symptoms associated with angiogenesis.
Cytotoxic agents are
2 0 numerous and varied and include, but are not limited to, cytotoxic drugs
or toxins or active fragments
of such toxins. Suitable toxins and their corresponding fragments include
diptheria A chain, exotoxin
A chain, ricin A chain, abrin A chain, curcin, crotin, phenomycin, enomycin
and the like. Cytotoxic
agents also include radiochemicals made by conjugating radioisotopes to
antibodies raised against
angiogenesis proteins, or binding of a radionuclide to a chelating agent that
has been covalently
2 5 attached to the antibody. Targeting the therapeutic moiety to
transmembrane angiogenesis proteins
not only serves to increase the local concentration of therapeutic moiety in
the angiogenesis afflicted
area, but also serves to reduce deleterious side effects that may be
associated with the therapeutic
moiety.
In another preferred embodiment, the angiogenesis protein against which the
antibodies are raised is
3 0 an intracellular protein. In this case, the antibody may be conjugated to
a protein which facilitates
entry into the cell. In one case, the antibody enters the cell by endocytosis.
In another embodiment, a
nucleic acid encoding the antibody is administered to the individual or cell.
Moreover, wherein the
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angiogenesis protein can be targeted within a cell, i.e., the nucleus, an
antibody thereto contains a
signal for that target localization, i.e., a nuclear localization signal.
The angiogenesis antibodies of the invention specifically bind to angiogenesis
proteins. By
"specifically bind" herein is meant that the antibodies bind to the protein
with a binding constant in the
range of at least 10~'- 10-6 M'', with a preferred range being 10-' - 10-9 M-
'.
In a preferred embodiment, the angiogenesis protein is purified or isolated
after expression.
Angiogenesis proteins may be isolated or purified in a variety of ways known
to those skilled in the art
depending on what other components are present in the sample. Standard
purification methods
include electrophoretic, molecular, immunological and chromatographic
techniques, including ion
exchange, hydrophobic, affinity, and reverse-phase HPLC chromatography, and
chromatofocusing.
For example, the angiogenesis protein may be purified using a standard anti-
angiogenesis protein
antibody column. Ultrafiltration and diafiltration techniques, in conjunction
with protein concentration,
are also useful. For general guidance in suitable purification techniques, see
Scopes, R., Protein
Purification, Springer-Verlag, NY (1982). The degree of purification necessary
will vary depending on
the use of the angiogenesis protein. In some instances no purification will be
necessary.
Once expressed and purified if necessary, the angiogenesis proteins and
nucleic acids are useful in a
number of applications.
In one aspect, the expression levels of genes are determined for different
cellular states in the
angiogenesis phenotype; that is, the expression levels of genes in normal
tissue (i.e. not undergoing
2 0 angiogenesis) and in angiogenesis tissue (and in some cases, for varying
severities of angiogenesis
that relate to prognosis, as outlined below) are evaluated to provide
expression profiles. An
expression profile of a particular cell state or point of development is
essentially a "fingerprint" of the
state; while two states may have any particular gene similarly expressed, the
evaluation of a number
of genes simultaneously allows the generation of a gene expression profile
that is unique to the state
2 5 of the cell. By comparing expression profiles of cells in different
states, information regarding which
genes are important (including both up- and down-regulation of genes) in each
of these states is
obtained. Then, diagnosis may be done or confirmed: does tissue from a
particular patient have the
gene expression profile of normal or angiogenesis tissue.
"Differential expression," or grammatical equivalents as used herein, refers
to both qualitative as well
3 0 as quantitative differences in the genes' temporal and/or cellular
expression patterns within and
among the cells. Thus, a differentially expressed gene can qualitatively have
its expression altered,
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including an activation or inactivation, in, for example, normal versus
angiogenic tissue. That is, genes
may be turned on or turned off in a particular state, relative to another
state. As is apparent to the
skilled artisan, any comparison of two or more states can be made. Such a
qualitatively regulated
gene will exhibit an expression pattern within a state or cell type which is
detectable by standard
techniques in one such state or cell type, but is not detectable in both.
Alternatively, the determination
is quantitative in that expression is increased or decreased; that is, the
expression of the gene is either
upregulated, resulting in an increased amount of transcript, or downregulated,
resulting in a decreased
amount of transcript. The degree to which expression differs need only be
large enough to quantify
via standard characterization techniques as outlined below, such as by use of
Affymetrix GeneChipT""
expression arrays, Lockhart, Nature Biotechnology, 14:1675-1680 (1996), hereby
expressly
incorporated by reference. Other techniques include, but are not limited to,
quantitative reverse
transcriptase PCR, Northern analysis and RNase protection. As outlined above,
preferably the change
in expression (i.e. upregulation or downregulation) is at least about 50%,
more preferably at least
about 100%, more preferably at least about 150%, more preferably, at least
about 200%, with from
300 to at least 1000% being especially preferred.
As will be appreciated by those in the art, this may be done by evaluation at
either the gene transcript,
or the protein level; that is, the amount of gene expression may be monitored
using nucleic acid
probes to the DNA or RNA equivalent of the gene transcript, and the
quantification of gene expression
levels, or, alternatively, the final gene product itself (protein) can be
monitored, for example through
2 0 the use of antibodies to the angiogenesis protein and standard
immunoassays (ELISAs, etc.) or other
techniques, including mass spectroscopy assays, 2D gel electrophoresis assays,
etc. Thus, the
proteins corresponding to angiogenesis genes, i.e. those identified as being
important in an
angiogenesis phenotype, can be evaluated in an angiogenesis diagnostic test.
In a preferred embodiment, gene expression monitoring is done and a number of
genes, i.e. an
expression profile, is monitored simultaneously, although multiple protein
expression monitoring can
be done as well. Similarly, these assays may be done on an individual basis as
well.
In this embodiment, the angiogenesis nucleic acid probes are attached to
biochips as outlined herein
for the detection and quantification of angiogenesis sequences in a particular
cell. The assays are
further described below in the example.
3 0 In a preferred embodiment nucleic acids encoding the angiogenesis protein
are detected. Although
DNA or RNA encoding the angiogenesis protein may be detected, of particular
interest are methods
wherein the mRNA encoding an angiogenesis protein is detected. The presence of
mRNA in a
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sample is an indication that the angiogenesis gene has been transcribed to
form the mRNA, and
suggests that the protein is expressed. Probes to detect the mRNA can be any
nucleotide/deoxynucleotide probe that is complementary to and base pairs with
the mRNA and
includes but is not limited to oligonucleotides, cDNA or RNA. Probes also
should contain a detectable
label, as defined herein. In one method the mRNA is detected after
immobilizing the nucleic acid to be
examined on a solid support such as nylon membranes and hybridizing the probe
with the sample.
Following washing to remove the non-specifically bound probe, the label is
detected. In another
method detection of the mRNA is performed in situ. In this method
permeabilized cells or tissue
samples are contacted with a detectably labeled nucleic acid probe for
sufficient time to allow the
probe to hybridize with the target mRNA. Following washing to remove the non-
specifically bound
probe, the label is detected. For example a digoxygenin labeled riboprobe (RNA
probe) that is
complementary to the mRNA encoding an angiogenesis protein is detected by
binding the digoxygenin
with an anti-digoxygenin secondary antibody and developed with vitro blue
tetrazolium and
5-bromo-4-chloro-3-indoyl phosphate.
In a preferred embodiment, any of the three classes of proteins as described
herein (secreted,
transmembrane or intracellular proteins) are used in diagnostic assays. The
angiogenesis proteins,
antibodies, nucleic acids, modified proteins and cells containing angiogenesis
sequences are used in
diagnostic assays. This can be done on an individual gene or corresponding
polypeptide level. In a
preferred embodiment, the expression profiles are used, preferably in
conjunction with high throughput
2 0 screening techniques to allow monitoring for expression profile genes
and/or corresponding
polypeptides.
As described and defined herein, angiogenesis proteins, including
intracellular, transmembrane or
secreted proteins, find use as markers of angiogenesis. Detection of these
proteins in putative
angiogenesis tissue or patients allows for a determination or diagnosis of
angiogenesis. Numerous
2 5 methods known to those of ordinary skill in the art find use in detecting
angiogenesis. In one
embodiment, antibodies are used to detect angiogenesis proteins. A preferred
method separates
proteins from a sample or patient by electrophoresis on a gel (typically a
denaturing and reducing
protein gel, but may be any other type of gel including isoelectric focusing
gels and the like). Following
separation of proteins, the angiogenesis protein is detected by immunoblotting
with antibodies raised
3 0 against the angiogenesis protein. Methods of immunoblotting are well known
to those of ordinary skill
in the art.
In another preferred method, antibodies to the angiogenesis protein find use
in in situ imaging
techniques. In this method cells are contacted with from one to many
antibodies to the angiogenesis
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protein(s). Following washing to remove non-specific antibody binding, the
presence of the antibody
or antibodies is detected. In one embodiment the antibody is detected by
incubating with a secondary
antibody that contains a detectable label. In another method the primary
antibody to the angiogenesis
proteins) contains a detectable label. In another preferred embodiment each
one of multiple primary
antibodies contains a distinct and detectable label. This method finds
particular use in simultaneous
screening for a plurality of angiogenesis proteins. As will be appreciated by
one of ordinary skill in the
art, numerous other histological imaging techniques are useful in the
invention.
In a preferred embodiment the label is detected in a fluorometer which has the
ability to detect and
distinguish emissions of different wavelengths. In addition, a fluorescence
activated cell sorter (FACS)
can be used in the method.
In another preferred embodiment, antibodies find use in diagnosing
angiogenesis from blood samples.
As previously described, certain angiogenesis proteins are
secreted/circulating molecules. Blood
samples, therefore, are useful as samples to be probed or tested for the
presence of secreted
angiogenesis proteins. Antibodies can be used to detect the angiogenesis by
any of the previously
described immunoassay techniques including ELISA, immunoblotting (Western
blotting),
immunoprecipitation, BIACORE technology and the like, as will be appreciated
by one of ordinary skill
in the art.
In a preferred embodiment, in situ hybridization of labeled angiogenesis
nucleic acid probes to tissue
arrays is done. For example, arrays of tissue samples, including angiogenesis
tissue and/or normal
2 0 tissue, are made. In situ hybridization as is known in the art can then be
done.
It is understood that when comparing the fingerprints between an individual
and a standard, the skilled
artisan can make a diagnosis as well as a prognosis. It is further understood
that the genes which
indicate the diagnosis may differ from those which indicate the prognosis.
In a preferred embodiment, the angiogenesis proteins, antibodies, nucleic
acids, modified proteins and
2 5 cells containing angiogenesis sequences are used in prognosis assays. As
above, gene expression
profiles can be generated that correlate to angiogenesis severity, in terms of
long term prognosis.
Again, this may be done on either a protein or gene level, with the use of
genes being preferred. As
above, the angiogenesis probes are attached to biochips for the detection and
quantification of
angiogenesis sequences in a tissue or patient. The assays proceed as outlined
above for diagnosis.
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In a preferred embodiment any of the three classes of proteins as described
herein are used in drug
screening assays. The angiogenesis proteins, antibodies, nucleic acids,
modified proteins and cells
containing angiogenesis sequences are used in drug screening assays or by
evaluating the effect of
drug candidates on a "gene expression profile" or expression profile of
polypeptides. In a preferred
embodiment, the expression profiles are used, preferably in conjunction with
high throughput
screening techniques to allow monitoring for expression profile genes after
treatment with a candidate
agent, Zlokarnik, et al., Science 279, 84-8 (1998), Heid, 1996 #69.
In a preferred embodiment, the angiogenesis proteins, antibodies, nucleic
acids, modified proteins and
cells containing the native or modified angiogenesis proteins are used in
screening assays. That is,
the present invention provides novel methods for screening for compositions
which modulate the
angiogenesis phenotype. As above, this can be done on an individual gene level
or by evaluating the
effect of drug candidates on a "gene expression profile". In a preferred
embodiment, the expression
profiles are used, preferably in conjunction with high throughput screening
techniques to allow
monitoring for expression profile genes after treatment with a candidate
agent, see Zlokarnik, supra.
Having identified the differentially expressed genes herein, a variety of
assays may be executed. In a
preferred embodiment, assays may be run on an individual gene or protein
level. That is, having
identified a particular gene as up regulated in angiogenesis, candidate
bioactive agents may be
screened to modulate this gene's response; preferably to down regulate the
gene, although in some
circumstances to up regulate the gene. "Modulation" thus includes both an
increase and a decrease
2 0 in gene expression. The preferred amount of modulation will depend on the
original change of the
gene expression in normal versus tissue undergoing angiogenesis, with changes
of at least 10%,
preferably 50%, more preferably 100-300%, and in some embodiments 300-1000% or
greater. Thus,
if a gene exhibits a 4 fold increase in angiogenic tissue compared to normal
tissue, a decrease of
about four fold is desired; a 10 fold decrease in angiogenic tissue compared
to normal tissue gives a
10 fold increase in expression for a candidate agent being desired.
As will be appreciated by those in the art, this may be done by evaluation at
either the gene or the
protein level; that is, the amount of gene expression may be monitored using
nucleic acid probes and
the quantification of gene expression levels, or, alternatively, the gene
product itself can be monitored,
for example through the use of antibodies to the angiogenesis protein and
standard immunoassays.
3 0 In a preferred embodiment, gene expression monitoring is done and a number
of genes, i.e. an
expression profile, is monitored simultaneously, although multiple protein
expression monitoring can
be done as well.
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In this embodiment, the angiogenesis nucleic acid probes are attached to
biochips as outlined herein
for the detection and quantification of angiogenesis sequences in a particular
cell. The assays are
further described below.
Generally, in a preferred embodiment, a candidate bioactive agent is added to
the cells prior to
analysis. Moreover, screens are provided to identify a candidate bioactive
agent which modulates
angiogenesis, modulates angiogenesis proteins, binds to an angiogenesis
protein, or interferes
between the binding of an angiogenesis protein and an antibody.
The term "candidate bioactive agent" or "drug candidate" or grammatical
equivalents as used herein
describes any molecule, e.g., protein, oligopeptide, small organic molecule,
polysaccharide,
polynucleotide, etc., to be tested for bioactive agents that are capable of
directly or indirectly altering
either the angiogenesis phenotype or the expression of an angiogenesis
sequence, including both
nucleic acid sequences and protein sequences. In preferred embodiments, the
bioactive agents
modulate the expression profiles, or expression profile nucleic acids or
proteins provided herein. In a
particularly preferred embodiment, the candidate agent suppresses an
angiogenesis phenotype, for
example to a normal tissue fingerprint. Similarly, the candidate agent
preferably suppresses a severe
angiogenesis phenotype. Generally a plurality of assay mixtures are run in
parallel with different agent
concentrations to obtain a differential response to the various
concentrations. Typically, one of these
concentrations serves as a negative control, i.e., at zero concentration or
below the level of detection.
In one aspect, a candidate agent will neutralize the effect of an angiogenesis
protein. By "neutralize"
2 0 is meant that activity of a protein is either inhibited or counter acted
against so as to have substantially
no effect on a cell.
Candidate agents encompass numerous chemical classes, though typically they
are organic
molecules, preferably small organic compounds having a molecular weight of
more than 100 and less
than about 2,500 daltons. Preferred small molecules are less than 2000, or
less than 1500 or less
2 5 than 1000 or less than 500 D. Candidate agents comprise functional groups
necessary for structural
interaction with proteins, particularly hydrogen bonding, and typically
include at least an amine,
carbonyl, hydroxyl or carboxyl group, preferably at least two of the
functional chemical groups. The
candidate agents often comprise cyclical carbon or heterocyclic structures
and/or aromatic or
polyaromatic structures substituted with one or more of the above functional
groups. Candidate
3 0 agents are also found among biomolecules including peptides, saccharides,
fatty acids, steroids,
purines, pyrimidines, derivatives, structural analogs or combinations thereof.
Particularly preferred are
peptides.
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Candidate agents are obtained from a wide variety of sources including
libraries of synthetic or natural
compounds. For example, numerous means are available for random and directed
synthesis of a
wide variety of organic compounds and biomolecules, including expression of
randomized
oligonucleotides. Alternatively, libraries of natural compounds in the form of
bacterial, fungal, plant
and animal extracts arE; available or readily produced. Additionally, natural
or synthetically produced
libraries and compounds are readily modified through conventional chemical,
physical and
biochemical means. Known pharmacological agents may be subjected to directed
or random
chemical modifications, such as acylation, alkylation, esterification,
amidification to produce structural
analogs.
In a preferred embodiment, the candidate bioactive agents are proteins. By
"protein" herein is meant
at least two covalently attached amino acids, which includes proteins,
polypeptides, oligopeptides and
peptides. The protein may be made up of naturally occurring amino acids and
peptide bonds, or
synthetic peptidomimetic structures. Thus "amino acid", or "peptide residue",
as used herein means
both naturally occurring and synthetic amino acids. For example, homo-
phenylalanine, citrulline and
noreleucine are considered amino acids for the purposes of the invention.
"Amino acid" also includes
imino acid residues such as proline and hydroxyproline. The side chains may be
in either the (R) or
the (S) configuration. In the preferred embodiment, the amino acids are in the
(S) or L-configuration.
If non-naturally occurring side chains are used, non-amino acid substituents
may be used, for example
to prevent or retard in vivo degradations.
2 0 In a preferred embodiment, the candidate bioactive agents are naturally
occurring proteins or
fragments of naturally occurring proteins. Thus, for example, cellular
extracts containing proteins, or
random or directed digests of proteinaceous cellular extracts, may be used. In
this way libraries of
procaryotic and eucaryotic proteins may be made for screening in the methods
of the invention.
Particularly preferred in this embodiment are libraries of bacterial, fungal,
viral, and mammalian
2 5 proteins, with the latter being preferred, and human proteins being
especially preferred.
In a preferred embodiment, the candidate bioactive agents are peptides of from
about 5 to about 30
amino acids, with from about 5 to about 20 amino acids being preferred, and
from about 7 to about 15
being particularly preferred. The peptides may be digests of naturally
occurring proteins as is outlined
above, random peptides, or "biased" random peptides. By "randomized" or
grammatical equivalents
3 0 herein is meant that each nucleic acid and peptide consists of essentially
random nucleotides and
amino acids, respectively. Since generally these random peptides (or nucleic
acids, discussed below)
are chemically synthesized, they may incorporate any nucleotide or amino acid
at any position. The
synthetic process can be designed to generate randomized proteins or nucleic
acids, to allow the
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formation of all or most of the possible combinations over the length of the
sequence, thus forming a
library of randomized candidate bioactive proteinaceous agents.
In one embodiment, the library is fully randomized, with no sequence
preferences or constants at any
position. In a preferred embodiment, the library is biased. That is, some
positions within the
sequence are either held constant, or are selected from a limited number of
possibilities. For
example, in a preferred embodiment, the nucleotides or amino acid residues are
randomized within a
defined class, for example, of hydrophobic amino acids, hydrophilic residues,
sterically biased (either
small or large) residues, towards the creation of nucleic acid binding
domains, the creation of
cysteines, for cross-linking, prolines for SH-3 domains, serines, threonines,
tyrosines or histidines for
phosphorylation sites, etc., or to purines, etc.
In a preferred embodiment, the candidate bioactive agents are nucleic acids,
as defined above.
As described above generally for proteins, nucleic acid candidate bioactive
agents may be naturally
occurring nucleic acids, random nucleic acids, or "biased" random nucleic
acids. For example, digests
of procaryotic or eucaryotic genomes may be used as is outlined above for
proteins.
In a preferred embodiment, the candidate bioactive agents are organic chemical
moieties, a wide
variety of which are available in the literature.
After the candidate agent has been added and the cells allowed to incubate for
some period of time,
the sample containing the target sequences to be analyzed is added to the
biochip. If required, the
target sequence is prepared using known techniques. For example, the sample
may be treated to
2 0 lyse the cells, using known lysis buffers, electroporation, etc., with
purification and/or amplification
such as PCR occurring as needed, as will be appreciated by those in the art.
For example, an in vitro
transcription with labels covalently attached to the nucleosides is done.
Generally, the nucleic acids
are labeled with biotin-FITC or PE, or with cy3 or cy5.
In a preferred embodiment, the target sequence is labeled with, for example, a
fluorescent, a
2 5 chemiluminescent, a chemical, or a radioactive signal, to provide a means
of detecting the target
sequence's specific binding to a probe. The label also can be an enzyme, such
as, alkaline
phosphatase or horseradish peroxidase, which when provided with an appropriate
substrate produces
a product that can be detected. Alternatively, the label can be a labeled
compound or small molecule,
such as an enzyme inhibitor, that binds but is not catalyzed or altered by the
enzyme. The label also
3 0 can be a moiety or compound, such as, an epitope tag or biotin which
specifically binds to streptavidin.
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For the example of biotin, the streptavidin is labeled as described above,
thereby, providing a
detectable signal for the bound target sequence. As known in the art, unbound
labeled streptavidin is
removed prior to analysis.
As will be appreciated by those in the art, these assays can be direct
hybridization assays or can
comprise "sandwich assays", which include the use of multiple probes, as is
generally outlined in U.S.
Patent Nos. 5,681,702, 5,597,909, 5,545,730, 5,594,117, 5,591,584, 5,571,670,
5,580,731, 5,571,670,
5,591,584, 5,624,802, 5,635,352, 5,594,118, 5,359,100, 5,124,246 and
5,681,697, all of which are
hereby incorporated by reference. In this embodiment, in general, the target
nucleic acid is prepared
as outlined above, and then added to the biochip comprising a plurality of
nucleic acid probes, under
conditions that allow the formation of a hybridization complex.
A variety of hybridization conditions may be used in the present invention,
including high, moderate
and low stringency conditions as outlined above. The assays are generally run
under stringency
conditions which allows formation of the label probe hybridization complex
only in the presence of
target. Stringency can be controlled by altering a step parameter that is a
thermodynamic variable,
including, but not limited to, temperature, formamide concentration, salt
concentration, chaotropic s2!t
concentration pH, organic solvent concentration, etc.
These parameters may also be used to control non-specific binding, as is
generally outlined in U.S.
Patent No. 5,681,697. Thus it may be desirable to perform certain steps at
higher stringency
conditions to reduce non-specific binding.
2 0 The reactions outlined herein may be accomplished in a variety of ways, as
will be appreciated by
those in the art. Components of the reaction may be added simultaneously, or
sequentially, in any
order, with preferred embodiments outlined below. In addition, the reaction
may include a variety of
other reagents may be included in the assays. These include reagents like
salts, buffers, neutral
proteins, e.g. albumin, detergents, etc which may be used to facilitate
optimal hybridization and
2 5 detection, and/or reduce non-specific or background interactions. Also
reagents that otherwise
improve the efficiency of the assay, such as protease inhibitors, nuclease
inhibitors, anti-microbial
agents, etc., may be used, depending on the sample preparation methods and
purity of the target.
Once the assay is run, the data is analyzed to determine the expression
levels, and changes in
expression levels as between states, of individual genes, forming a gene
expression profile.
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The screens are done to identify drugs or bioactive agents that modulate the
angiogenesis phenotype.
Specifically, there are several types of screens that can be run. A preferred
embodiment is in the
screening of candidate agents that can induce or suppress a particular
expression profile, thus
preferably generating the associated phenotype. That is, candidate agents that
can mimic or produce
an expression profile in angiogenesis similar to the expression profile of
normal tissue is expected to
result in a suppression of the angiogenesis phenotype. Thus, in this
embodiment, mimicking an
expression profile, or changing one profile to another, is the goal.
In a preferred embodiment, as for the diagnosis applications, having
identified the differentially
expressed genes important in any one state, screens can be run to alter the
expression of the genes
individually. That is, screening for modulation of regulation of expression of
a single gene can be
done; that is, rather than try to mimic all or part of an expression profile,
screening for regulation of
individual genes can be done. Thus, for example, particularly in the case of
target genes whose
presence or absence is unique between two states, screening is done for
modulators of the target
gene expression.
In a preferred embodiment, screening is done to alter the biological function
of the expression product
of the differentially expressed gene. Again, having identified the importance
of a gene in a particular
state, screening for agents that bind andlor modulate the biological activity
of the gene product can be
run as is more fully outlined below.
Thus, screening of candidate agents that modulate the angiogenesis phenotype
either at the gene
2 0 expression level or the protein level can be done.
In addition screens can be done for novel genes that are induced in response
to a candidate agent.
After identifying a candidate agent based upon its ability to suppress an
angiogenesis expression
pattern leading to a normal expression pattern, or modulate a single
angiogenesis gene expression
profile so as to mimic the expression of the gene from normal tissue, a screen
as described above can
2 5 be performed to identify genes that are specifically modulated in response
to the agent. Comparing
expression profiles between normal tissue and agent treated angiogenesis
tissue reveals genes that
are not expressed in normal tissue or angiogenesis tissue, but are expressed
in agent treated tissue.
These agent specific sequences can be identified and used by any of the
methods described herein
for angiogenesis genes or proteins. In particular these sequences and the
proteins they encode find
3 0 use in marking or identifying agent treated cells. In addition, antibodies
can be raised against the
agent induced proteins and used to target novel therapeutics to the treated
angiogenesis tissue
sample.
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Thus, in one embodiment, a candidate agent is administered to a population of
angiogenic cells, that
thus has an associated angiogenesis expression profile. By "administration" or
"contacting" herein is
meant that the candidate agent is added to the cells in such a manner as to
allow the agent to act
upon the cell, whether by uptake and intracellular action, or by action at the
cell surface. In some
embodiments, nucleic acid encoding a proteinaceous candidate agent (i.e. a
peptide) may be put into
a viral construct such as a retroviral construct and added to the cell, such
that expression of the
peptide agent is accomplished; see PCT US97/01019, hereby expressly
incorporated by reference.
Once the candidate agent has been administered to the cells, the cells can be
washed if desired and
are allowed to incubate under preferably physiological conditions for some
period of time. The cells
are then harvested and a new gene expression profile is generated, as outlined
herein.
Thus, for example, angiogenesis tissue may be screened for agents that reduce
or suppress the
angiogenesis phenotype. A change in at least one gene of the expression
profile indicates that the
agent has an effect on angiogenesis activity. By defining such a signature for
the angiogenesis
phenotype, screens for new drugs that alter the phenotype can be devised. With
this approach, the
drug target need not be known and need not be represented in the original
expression screening
platform, nor does the level of transcript for the target protein need to
change.
In a preferred embodiment, as outlined above, screens may be done on
individual genes and gene
products (proteins). That is, having identified a particular differentially
expressed gene as important in
a particular state, screening of modulators of either the expression of the
gene or the gene product
2 0 itself can be done. The gene products of differentially expressed genes
are sometimes referred to
herein as "angiogenesis proteins". In preferred embodiments the angiogenesis
protein is as depicted
in Figures 4, 8, 13, 18, and 22 or encoded by the sequences shown in figures
2, 3, 7, 12, 17, 21 and
23. The angiogenesis protein may be a fragment, or alternatively, be the full
length protein to a
fragment shown herein.
2 5 Preferably, the angiogenesis protein is a fragment of approximately 14 to
24 amino acids long. More
preferably the fragment is a soluble fragment.
In a preferred embodiment, the fragment is from AAA1. Preferably, the fragment
includes a non-
transmembrane region. In a preferred embodiment, the AAA1 fragment has an N-
terminal Cys to aid
in solubility. Preferably, the fragment is selected from AAA1 p1 and AAA1 p2.
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In a preferred embodiment, the fragment is charged and from the c-terminus of
AAA4. In one
embodiment, the c-terminus of the fragment is kept as a free acid and the n-
terminus is a free amine
to aid in coupling, i.e., to cysteine. In one embodiment the fragment is an
internal peptide overlapping
hydrophilic stretch of AAA4. In a preferred embodiment, the termini is
blocked. Preferably, the
fragment of AAA4 is selected from AAA4p1 or AAA4p2. In another preferred
embodiment, the
fragment is a novel fragment from the N-terminal. In one embodiment, the
fragment excludes
sequence outside of the N-terminal, in another embodiment, the fragment
includes at least a portion of
the N-terminal. "N-terminal" is used interchangeably herein with "N-terminus"
which is further
described above.
In one embodiment the angiogenesis proteins are conjugated to an immunogenic
agent as discussed
herein. In one embodiment the angiogenesis protein is conjugated to BSA.
Thus, in a preferred embodiment, screening for modulators of expression of
specific genes can be
done. This will be done as outlined above, but in general the expression of
only one or a few genes
are evaluated.
In a preferred embodiment, screens are designed to first find candidate agents
that can bind to
differentially expressed proteins, and then these agents may be used in assays
that evaluate the
ability of the candidate agent to modulate differentially expressed activity.
Thus, as will be appreciated
by those in the art, there are a number of different assays which may be run;
binding assays and
activity assays.
2 0 In a preferred embodiment, binding assays are done. In general, purified
or isolated gene product is
used; that is, the gene products of one or more differentially expressed
nucleic acids are made. In
general, this is done as is known in the art. For example, antibodies are
generated to the protein gene
products, and standard immunoassays are run to determine the amount of protein
present.
Alternatively, cells comprising the angiogenesis proteins can be used in the
assays.
2 5 Thus, in a preferred embodiment, the methods comprise combining an
angiogenesis protein and a
candidate bioactive agent, and determining the binding of the candidate agent
to the angiogenesis
protein. Preferred embodiments utilize the human angiogenesis protein,
although other mammalian
proteins may also be used, for example for the development of animal models of
human disease. In
some embodiments, as outlined herein, variant or derivative angiogenesis
proteins may be used.
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Generally, in a preferred embodiment of the methods herein, the angiogenesis
protein or the
candidate agent is non-diffusably bound to an insoluble support having
isolated sample receiving
areas (e.g. a microtiter plate, an array, etc.). The insoluble supports may be
made of any composition
to which the compositions can be bound, is readily separated from soluble
material, and is otherwise
compatible with the overall method of screening. The surface of such supports
may be solid or porous
and of any convenient shape. Examples of suitable insoluble supports include
microtiter plates,
arrays, membranes and beads. These are typically made of glass, plastic (e.g.,
polystyrene),
polysaccharides, nylon or nitrocellulose, teflonT"', etc. Microtiter plates
and arrays are especially
convenient because a large number of assays can be carried out simultaneously,
using small amounts
of reagents and samples. The particular manner of binding of the composition
is not crucial so long
as it is compatible with the reagents and overall methods of the invention,
maintains the activity of the
composition and is nondiffusable. Preferred methods of binding include the use
of antibodies (which
do not sterically block either the ligand binding site or activation sequence
when the protein is bound to
the support), direct binding to "sticky" or ionic supports, chemical
crosslinking, the synthesis of the
protein or agent on the surface, etc. Following binding of the protein or
agent, excess unbound
material is removed by washing. The sample receiving areas may then be blocked
through incubation
with bovine serum albumin (BSA), casein or other innocuous protein or other
moiety.
In a preferred embodiment, the angiogenesis protein is bound to the support,
and a candidate
bioactive agent is added to the assay. Alternatively, the candidate agent is
bound to the support and
2 0 the angiogenesis protein is added. Novel binding agents include specific
antibodies, non-natural
binding agents identified in screens of chemical libraries, peptide analogs,
etc. Of particular interest
are screening assays for agents that have a low toxicity for human cells. A
wide variety of assays may
be used for this purpose, including labeled in vitro protein-protein binding
assays, electrophoretic
mobility shift assays, immunoassays for protein binding, functional assays
(phosphorylation assays,
etc.) and the like.
The determination of the binding of the candidate bioactive agent to the
angiogenesis protein may be
done in a number of ways. In a preferred embodiment, the candidate bioactive
agent is labelled, and
binding determined directly. For example, this may be done by attaching all or
a portion of the
angiogenesis protein to a solid support, adding a labelled candidate agent
(for example a fluorescent
3 0 label), washing off excess reagent, and determining whether the label is
present on the solid support.
Various blocking and washing steps may be utilized as is known in the art.
By "labeled" herein is meant that the compound is either directly or
indirectly labeled with a label which
provides a detectable signal, e.g. radioisotope, fluorescers, enzyme,
antibodies, particles such as
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magnetic particles, chemiluminescers, or specific binding molecules, etc.
Specific binding molecules
include pairs, such as biotin and streptavidin, digoxin and antidigoxin etc.
For the specific binding
members, the complementary member would normally be labeled with a molecule
which provides for
detection, in accordance with known procedures, as outlined above. The label
can directly or indirectly
provide a detectable signal.
In some embodiments, only one of the components is labeled. For example, the
proteins (or
proteinaceous candidate agents) may be labeled at tyrosine positions using
'251, or with fluorophores.
Alternatively, more than one component may be labeled with different labels;
using'z51 for the proteins,
for example, and a fluorophor for the candidate agents.
In a preferred embodiment, the binding of the candidate bioactive agent is
determined through the use
of competitive binding assays. In this embodiment, the competitor is a binding
moiety known to bind
to the target molecule (i.e. angiogenesis), such as an antibody, peptide,
binding partner, ligand, etc.
Under certain circumstances, there may be competitive binding as between the
bioactive agent and
the binding moiety, with the binding moiety displacing the bioactive agent.
In one embodiment, the candidate bioactive agent is labeled. Either the
candidate bioactive agent, or
the competitor, or both, is added first to the protein for a time sufficient
to allow binding, if present.
Incubations may be performed at any temperature which facilitates optimal
activity, typically between 4
and 40°C. Incubation periods are selected for optimum activity, but may
also be optimized to facilitate
rapid high through put screening. Typically between 0.1 and 1 hour will be
sufficient. Excess reagent
2 0 is generally removed or washed away. The second component is then added,
and the presence or
absence of the labeled component is followed, to indicate binding.
In a preferred embodiment, the competitor is added first, followed by the
candidate bioactive agent.
Displacement of the competitor is an indication that the candidate bioactive
agent is binding to the
angiogenesis protein and thus is capable of binding to, and potentially
modulating, the activity of the
2 5 angiogenesis protein. In this embodiment, either component can be labeled.
Thus, for example, if the
competitor is labeled, the presence of label in the wash solution indicates
displacement by the agent.
Alternatively, if the candidate bioactive agent is labeled, the presence of
the label on the support
indicates displacement.
In an alternative embodiment, the candidate bioactive agent is added first,
with incubation and
3 0 washing, followed by the competitor. The absence of binding by the
competitor may indicate that the
bioactive agent is bound to the angiogenesis protein with a higher affinity.
Thus, if the candidate
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bioactive agent is labeled, the presence of the label on the support, coupled
with a lack of competitor
binding, may indicate that the candidate agent is capable of binding to the
angiogenesis protein.
In a preferred embodiment, the methods comprise differential screening to
identity bioactive agents
that are capable of modulating the activitity of the angiogenesis proteins. In
this embodiment, the
methods comprise combining an angiogenesis protein and a competitor in a first
sample. A second
sample comprises a candidate bioactive agent, an angiogenesis protein and a
competitor. The
binding of the competitor is determined for both samples, and a change, or
difference in binding
between the two samples indicates the presence of an agent capable of binding
to the angiogenesis
protein and potentially modulating its activity. That is, if the binding of
the competitor is different in the
second sample relative to the first sample, the agent is capable of binding to
the angiogenesis protein.
Alternatively, a preferred embodiment utilizes differential screening to
identify drug candidates that
bind to the native angiogenesis protein, but cannot bind to modified
angiogenesis proteins. The
structure of the angiogenesis protein may be modeled, and used in rational
drug design to synthesize
agents that interact with that site. Drug candidates that affect angiogenesis
bioactivity are also
identified by screening drugs for the ability to either enhance or reduce the
activity of the protein.
Positive controls and negative controls may be used in the assays. Preferably
all control and test
samples are performed in at least triplicate to obtain statistically
significant results. Incubation of all
samples is for a time sufficient for the binding of the agent to the protein.
Following incubation, all
samples are washed free of non-specifically bound material and the amount of
bound, generally
2 0 labeled agent determined. For example, where a radiolabel is employed, the
samples may be
counted in a scintillation counter to determine the amount of bound compound.
A variety of other reagents may be included in the screening assays. These
include reagents like
salts, neutral proteins, e.g. albumin, detergents, etc which may be used to
facilitate optimal
protein-protein binding and/or reduce non-specific or background interactions.
Also reagents that
2 5 otherwise improve the efficiency of the assay, such as protease
inhibitors, nuclease inhibitors,
anti-microbial agents, etc., may be used. The mixture of components may be
added in any order that
provides for the requisite binding.
Screening for agents that modulate the activity of angiogenesis proteins may
also be done. In a
preferred embodiment, methods for screening for a bioactive agent capable of
modulating the activity
3 0 of angiogenesis proteins comprise the steps of adding a candidate
bioactive agent to a sample of
angiogenesis proteins, as above, and determining an alteration in the
biological activity of
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angiogenesis proteins. "Modulating the activity of angiogenesis proteins"
includes an increase in
activity, a decrease in activity, or a change in the type or kind of activity
present. Thus, in this
embodiment, the candidate agent should both bind to angiogenesis
proteins(although this may not be
necessary), and alter its biological or biochemical activity as defined
herein. The methods include
both in vitro screening methods, as are generally outlined above, and in vivo
screening of cells for
alterations in the presence, distribution, activity or amount of angiogenesis
proteins.
Thus, in this embodiment, the methods comprise combining an angiogenesis
sample and a candidate
bioactive agent, and evaluating the effect on angiogenesis. By "angiogenesis
activity" or grammatical
equivalents herein is meant one of angiogenesis's biological activities,
including, but not limited to, its
role in angiogenesis. In one embodiment, angiogenesis activity includes
activation of AAA4, AAA1,
Edg-1, alpha 5 beta1 integrin, endomucin and matrix metalloproteinase 10. An
inhibitor of
angiogenesis activity is the inhibition of any one or more angiogenesis
activities.
In a preferred embodiment, the activity of the angiogenesis protein is
increased; in another preferred
embodiment, the activity of the angiogenesis protein is decreased. Thus,
bioactive agents that are
antagonists are preferred in some embodiments, and bioactive agents that are
agonists may be
preferred in other embodiments.
In a preferred embodiment, the invention provides methods for screening for
bioactive agents capable
of modulating the activity of an angiogenesis protein. The methods comprise
adding a candidate
bioactive agent, as defined above, to a cell comprising angiogenesis proteins.
Preferred cell types
2 0 include almost any cell. The cells contain a recombinant nucleic acid that
encodes an angiogenesis
protein. In a preferred embodiment, a library of candidate agents are tested
on a plurality of cells.
In one aspect, the assays are evaluated in the presence or absence or previous
or subsequent
exposure of physiological signals, for example hormones, antibodies, peptides,
antigens, cytokines,
growth factors, action potentials, pharmacological agents including
chemotherapeutics, radiation,
2 5 carcinogenics, or other cells (i.e. cell-cell contacts). In another
example, the determinations are
determined at different stages of the cell cycle process.
In this way, bioactive agents are identified. Compounds with pharmacological
activity are able to
enhance or interfere with the activity of the angiogenesis protein. In one
embodiment, "angiogenesis
protein activity" as used herein includes at least one of the following:
angiogenesis protein activity as
3 0 defined herein, binding to Edg-1, activation of Edg-1, or activation of
substrates of Edg-1. In one
embodiment, angiogenesis activity is defined as the unregulated proliferation
of angiogenic tissue, or
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the growth of arteries in tissue. In one aspect, angiogenesis activity as
defined herein is related to the
activity of Edg-1 in the upregulation of Edg-1 in angiogenic tissue.
In another embodiment, angiogenesis protein activity includes at least one of
the following:
angiogenesis activity, binding to one of AAA4, AAA1, Edg-1, alpha 5 beta 1
integrin, endomucin,
matrix metalloproteinase 10, or activation of substrates of AAA4, AAA1, Edg-1,
alpha 5 beta 1 integrin,
endomucin, matrix metalloproteinase 10, respectively. In one preferred
embodiment, AAA1 comprises
its N-terminal end. In one aspect, angiogenesis activity as defined herein is
related to the activity of
AAA4, AAA1, Edg-1, alpha 5 beta 1 integrin, endomucin, matrix
metalloproteinase 10, in the
upregulation of AAA4, AAA1, Edg-1, alpha 5 beta 1 integrin, endomucin, matrix
metalloproteinase 10,
respectively in angiogenesis tissue.
In one embodiment, a method of inhibiting angiogenic cell division is
provided. The method comprises
administration of a angiogenesis inhibitor.
In another embodiment, a method of inhibiting angiogenesis is provided. The
method comprises
administration of an angiogenesis inhibitor.
In a further embodiment, methods of treating cells or individuals with
angiogenesis are provided. The
method comprises administration of an angiogenesis inhibitor.
In one embodiment, an angiogenesis inhibitor is an antibody as discussed
above. In another
embodiment, the angiogenesis inhibitor is an antisense molecule. Antisense
molecules as used
herein include antisense or sense oligonucleotides comprising a singe-stranded
nucleic acid sequence
2 0 (either RNA or DNA) capable of binding to target mRNA (sense) or DNA
(antisense) sequences for
angiogenesis molecules. A preferred antisense molecule is for AAA4, AAA1, Edg-
1, alpha 5 beta 1
integrin, endomucin, or matrix metalloproteinase 10, more preferable the
angiogenesis sequences in
Table 5, or for a ligand or activator thereof. A most preferred antisense
molecule is for Edg-1 or for a
ligand or activator thereof. Antisense or sense oligonucleotides, according to
the present invention,
2 5 comprise a fragment generally at least about 14 nucleotides, preferably
from about 14 to 30
nucleotides. The ability to derive an antisense or a sense oligonucleotide,
based upon a cDNA
sequence encoding a given protein is described in, for example, Stein and
Cohen (Cancer Res.
48:2659, 1988) and van der Krol et al. (BioTechni4ues 6:958, 1988).
Antisense molecules may be introduced into a cell containing the target
nucleotide sequence by
3 0 formation of a conjugate with a ligand binding molecule, as described in
WO 91/04753. Suitable
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ligand binding molecules include, but are not limited to, cell surface
receptors, growth factors, other
cytokines, or other ligands that bind to cell surface receptors. Preferably,
conjugation of the ligand
binding molecule does not substantially interfere with the ability of the
ligand binding molecule to bind
to its corresponding molecule or receptor, or block entry of the sense or
antisense oligonucleotide or
its conjugated version into the cell. Alternatively, a sense or an antisense
oligonucleotide may be
introduced into a cell containing the target nucleic acid sequence by
formation of an oligonucleotide-
lipid complex, as described in WO 90/10448. It is understood that the use of
antisense molecules or
knock out and knock in models may also be used in screening assays as
discussed above, in addition
to methods of treatment.
The compounds having the desired pharmacological activity may be administered
in a physiologically
acceptable carrier to a host, as previously described. The agents may be
administered in a variety of
ways, orally, parenterally e.g., subcutaneously, intraperitoneally,
intravascularly, etc. Depending upon
the manner of introduction, the compounds may be formulated in a variety of
ways. The concentration
of therapeutically active compound in the formulation may vary from about 0.1-
100 wt.°/a. The agents
may be administered alone or in combination with other treatments, i.e.,
radiation.
The pharmaceutical compositions can be prepared in various forms, such as
granules, tablets, pills,
suppositories, capsules, suspensions, salves, lotions and the like.
Pharmaceutical grade organic or
inorganic carriers and/or diluents suitable for oral and topical use can be
used to make up
compositions containing the therapeutically-active compounds. Diluents known
to the art include
2 0 aqueous media, vegetable and animal oils and fats. Stabilizing agents,
wetting and emulsifying
agents, salts for varying the osmotic pressure or buffers for securing an
adequate pH value, and skin
penetration enhancers can be used as auxiliary agents.
Without being bound by theory, it appears that the various angiogenesis
sequences are important in
angiogenesis. Accordingly, disorders based on mutant or variant angiogenesis
genes may be
determined. In one embodiment, the invention provides methods for identifying
cells containing
variant angiogenesis genes comprising determining all or part of the sequence
of at least one
endogeneous angiogenesis genes in a cell. As will be appreciated by those in
the art, this may be
done using any number of sequencing techniques. In a preferred embodiment, the
invention provides
methods of identifying the angiogenesis genotype of an individual comprising
determining all or part of
3 0 the sequence of at least one angiogenesis gene of the individual. This is
generally done in at least
one tissue of the individual, and may include the evaluation of a number of
tissues or different samples
of the same tissue. The method may include comparing the sequence of the
sequenced angiogenesis
gene to a known angiogenesis gene, i.e. a wild-type gene.
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The sequence of all or part of the angiogenesis gene can then be compared to
the sequence of a
known angiogenesis gene to determine if any differences exist. This can be
done using any number
of known homology programs, such as Bestfit, etc. In a preferred embodiment,
the presence of a a
difference in the sequence between the angiogenesis gene of the patient and
the known angiogenesis
gene is indicative of a disease state or a propensity for a disease state, as
outlined herein.
In a preferred embodiment, the angiogenesis genes are used as probes to
determine the number of
copies of the angiogenesis gene in the genome.
In another preferred embodiment, the angiogenesis genes are used as probes to
determine the
chromosomal localization of the angiogenesis genes. Information such as
chromosomal localization
finds use in providing a diagnosis or prognosis in particular when chromosomal
abnormalities such as
translocations, and the like are identified in the angiogenesis gene locus.
Thus, in one embodiment, methods of modulating angiogenesis in cells or
organisms are provided. In
one embodiment, the methods comprise administering to a cell an anti-
angiogenesis antibody that
reduces or eliminates the biological activity of an endogeneous angiogenesis
protein. Alternatively,
the methods comprise administering to a cell or organism a recombinant nucleic
acid encoding an
angiogenesis protein. As will be appreciated by those in the art, this may be
accomplished in any
number of ways. In a preferred embodiment, for example when the angiogenesis
sequence is down-
regulated in angiogenesis, the activity of the angiogenesis gene is increased
by increasing the amount
of angiogenesis in the cell, for example by overexpressing the endogeneous
angiogenesis or by
2 0 administering a gene encoding the angiogenesis sequence, using known gene-
therapy techniques, for
example. In a preferred embodiment, the gene therapy techniques include the
incorporation of the
exogenous gene using enhanced homologous recombination (EHR), for example as
described in
PCT/US93/03868, hereby incorporated by reference in its entireity.
Alternatively, for example when
the angiogenesis sequence is up-regulated in angiogenesis, the activity of the
endogeneous
angiogenesis gene is decreased, for example by the administration of a
angiogenesis antisense
nucleic acid.
In one embodiment, the angiogenesis proteins of the present invention may be
used to generate
polyclonal and monoclonal antibodies to angiogenesis proteins, which are
useful as described herein.
Similarly, the angiogenesis proteins can be coupled, using standard
technology, to affinity
3 0 chromatography columns. These columns may then be used to purify
angiogenesis antibodies. In a
preferred embodiment, the antibodies are generated to epitopes unique to a
angiogenesis protein; that
is, the antibodies show little or no cross-reactivity to other proteins. These
antibodies find use in a
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number of applications. For example, the angiogenesis antibodies may be
coupled to standard affinity
chromatography columns and used to purify angiogenesis proteins. The
antibodies may also be used
as blocking polypeptides, as outlined above, since they will specifically bind
to the angiogenesis
protein.
In one embodiment, a therapeutically effective dose of an angiogenesis
proteins and modulator
thereof is administered to a patient. By "therapeutically effective dose"
herein is meant a dose that
produces the effects for which it is administered. The exact dose will depend
on the purpose of the
treatment, and will be ascertainable by one skilled in the art using known
techniques. As is known in
the art, adjustments for angiogenesis degradation, systemic versus localized
delivery, and rate of new
protease synthesis, as well as the age, body weight, general health, sex,
diet, time of administration,
drug interaction and the severity of the condition may be necessary, and will
be ascertainable with
routine experimentation by those skilled in the art.
A "patient" for the purposes of the present invention includes both humans and
other animals,
particularly mammals, and organisms. Thus the methods are applicable to both
human therapy and
veterinary applications. In the preferred embodiment the patient is a mammal,
and in the most
preferred embodiment the patient is human.
The administration of the angiogenesis proteins and modulators thereof of the
present invention can
be done in a variety of ways as discussed above, including, but not limited
to, orally, subcutaneously,
intravenously, intranasally, transdermally, intraperitoneally,
intramuscularly, intrapulmonary, vaginally,
2 0 rectally, or intraocularly. In some instances, for example, in the
treatment of wounds and
inflammation, the angiogenesis proteins and modulators may be directly applied
as a solution or spray.
The pharmaceutical compositions of the present invention comprise an
angiogenesis protein in a form
suitable for administration to a patient. In the preferred embodiment, the
pharmaceutical compositions
are in a water soluble form, such as being present as pharmaceutically
acceptable salts, which is
meant to include both acid and base addition salts. "Pharmaceutically
acceptable acid addition salt"
refers to those salts that retain the biological effectiveness of the free
bases and that are not
biologically or otherwise undesirable, formed with inorganic acids such as
hydrochloric acid,
hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like,
and organic acids such as
acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malefic
acid, malonic acid, succinic
3 0 acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic
acid, mandelic acid,
methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic
acid and the like.
"Pharmaceutically acceptable base addition salts" include those derived from
inorganic bases such as
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sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper,
manganese,
aluminum salts and the like. Particularly preferred are the ammonium,
potassium, sodium, calcium,
and magnesium salts. Salts derived from pharmaceutically acceptable organic
non-toxic bases
include salts of primary, secondary, and tertiary amines, substituted amines
including naturally
occurring substituted amines, cyclic amines and basic ion exchange resins,
such as isopropylamine,
trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine.
The pharmaceutical compositions may also include one or more of the following:
carrier proteins such
as serum albumin; buffers; fillers such as microcrystalline cellulose,
lactose, corn and other starches;
binding agents; sweeteners and other flavoring agents; coloring agents; and
polyethylene glycol.
Additives are well known in the art, and are used in a variety of
formulations.
In a preferred embodiment, angiogenesis proteins and modulators are
administered as therapeutic
agents, and can be formulated as outlined above. Similarly, angiogenesis genes
(including both the
full-length sequence, partial sequences, or regulatory sequences of the
angiogenesis coding regions)
can be administered in gene therapy applications, as is known in the art.
These angiogenesis genes
can include antisense applications, either as gene therapy (i.e. for
incorporation into the genome) or
as antisense compositions, as will be appreciated by those in the art.
In a preferred embodiment, angiogenesis genes are administered as DNA
vaccines, either single
genes or combinations of angiogenesis genes. Naked DNA vaccines are generally
known in the art.
Brower, Nature Biotechnology, 16:1304-1305 (1998).
2 0 In one embodiment, angiogenesis genes of the present invention are used as
DNA vaccines.
Methods for the use of genes as DNA vaccines are well known to one of ordinary
skill in the art, and
include placing an angiogenesis gene or portion of an angiogenesis gene under
the control of a
promoter for expression in an angiogenesis patient. The angiogenesis gene used
for DNA vaccines
can encode full-length angiogenesis proteins, but more preferably encodes
portions of the
angiogenesis proteins including peptides derived from the angiogenesis
protein. In a preferred
embodiment a patient is immunized with a DNA vaccine comprising a plurality of
nucleotide
sequences derived from an angiogenesis gene. Similarly, it is possible to
immunize a patient with a
plurality of angiogenesis genes or portions thereof as defined herein. Without
being bound by theory,
expression of the polypeptide encoded by the DNA vaccine, cytotoxic T-cells,
helper T-cells and
3 0 antibodies are induced which recognize and destroy or eliminate cells
expressing angiogenesis
proteins.
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In a preferred embodiment, the DNA vaccines include a gene encoding an
adjuvant molecule with the
DNA vaccine. Such adjuvant molecules include cytokines that increase the
immunogenic response to
the angiogenesis polypeptide encoded by the DNA vaccine. Additional or
alternative adjuvants are
known to those of ordinary skill in the art and find use in the invention.
In another preferred embodiment angiogenesis genes find use in generating
animal models of
angiogenesis. As is appreciated by one of ordinary skill in the art, when the
angiogenesis gene
identified is repressed or diminished in angiogenesis tissue, gene therapy
technology wherein
antisense RNA directed to the angiogenesis gene will also diminish or repress
expression of the gene.
An animal generated as such serves as an animal model of angiogenesis that
finds use in screening
bioactive drug candidates. Similarly, gene knockout technology, for example as
a result of
homologous recombination with an appropriate gene targeting vector, will
result in the absence of the
angiogenesis protein. When desired, tissue-specific expression or knockout of
the angiogenesis
protein may be necessary.
It is also possible that the angiogenesis protein is overexpressed in
angiogenesis. As such,
transgenic animals can be generated that overexpress the angiogenesis protein.
Depending on the
desired expression level, promoters of various strengths can be employed to
express the transgene.
Also, the number of copies of the integrated transgene can be determined and
compared for a
determination of the expression level of the transgene. Animals generated by
such methods find use
as animal models of angiogenesis and are additionally useful in screening for
bioactive molecules to
2 0 treat angiogenesis.
It is understood that the examples described above in no way serve to limit
the true scope of this
invention, but rather are presented for illustrative purposes. All references
and sequences of
accession numbers cited herein are incorporated by reference in their
entirety.
EXAMPLES
2 5 Example 1
Tissue Preparation, Labeling Chips, and Fingerprints
Purify total RNA from tissue using TRlzol Reagent
Estimate tissue weight. Homogenize tissue samples in 1 ml of TRlzol per 50mg
of tissue using a
Polytron 3100 homogenizer. The generator/probe used depends upon the tissue
size. A
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generator that is too large for the amount of tissue to be homogenized will
cause a loss of sample
and lower RNA yield. Use the 20mm generator for tissue weighing more than
0.6g. If the working
volume is greater than 2m1, then homogenize tissue in a 15m1 polypropylene
tube (Falcon 2059).
Fill tube no greater than 10m1.
HOMOGENIZATION
Before using generator, it should have been cleaned after last usage by
running it through soapy
H20 and rinsing thoroughly. Run through with EtOH to sterilize. Keep tissue
frozen until ready.
Add TRlzol directly to frozen tissue then homogenize.
Following homogenization, remove insoluble material from the homogenate by
centrifugation at
7500 x g for 15 min. in a Sorvall superspeed or 12,000 x g for 10 min. in an
Eppendorf centrifuge
at 4°C. Transfer the cleared homogenate to a new tube(s). The samples
may be frozen now at -
60 to -70°C (and kept for at least one month) or you may continue with
the purification.
PHASE SEPARATION
Incubate the homogenized samples for 5 minutes at room temperature.
Add 0.2m1 of chloroform per 1 ml of TRlzol reagent used in the original
homogenization.
Cap tubes securely and shake tubes vigorously by hand (do not vortex) for 15
seconds.
Incubate samples at room temp. for 2-3 minutes. Centrifuge samples at 6500rpm
in a Sorvall
superspeed for 30 min. at 4°C. (You may spin at up to 12,000 x g for 10
min. but you risk
breaking your tubes in the centrifuge.)
2 0 RNA PRECIPITATION
Transfer the aqueous phase to a fresh tube. Save the organic phase if
isolation of DNA or protein
is desired. Add 0.5m1 of isopropyl alcohol per 1 ml of TRlzol reagent used in
the original
homogenization. Cap tubes securely and invert to mix. Incubate samples at room
temp. for 10
minutes. Centrifuge samples at 6500rpm in Sorvall for 20min. at 4°C.
2 5 RNA WASH
Pour off the supernate. Wash pellet with cold 75% ethanol. Use 1 ml of 75%
ethanol per 1 ml of
TRlzol reagent used in the initial homogenization. Cap tubes securely and
invert several times to
loosen pellet. (Do not vortex). Centrifuge at <8000rpm (<7500 x g) for 5
minutes at 4°C.
Pour off the wash. Carefully transfer pellet to an eppendorf tube (let it
slide down the tube into the
3 0 new tube and use a pipet tip to help guide it in if necessary). Depending
on the volumes you are
working with, you can decide what size tubes) you want to precipitate the RNA
in. When I tried
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leaving the RNA in the large 15m1 tube, it took so long to dry (i.e. it did
not dry) that I eventually
had to transfer it to a smaller tube. Let pellet dry in hood. Resuspend RNA in
an appropriate
volume of DEPC H20. Try for 2-5ug/ul. Take absorbance readings.
Purify poly A+ mRNA from total RNA or clean up total RNA with Qiagen' s
RNeasy kit
Purification of poly A' mRNA from total RNA. Heat oligotex suspension to
37°C and mix
immediately before adding to RNA. Incubate Elution Buffer at 70°C. Warm
up 2 x Binding Buffer
at 65°C if there is precipitate in the buffer. Mix total RNA with DEPC-
treated water, 2 x Binding
Buffer, and Oligotex according to Table 2 on page 16 of the Oligotex Handbook.
Incubate for 3
minutes at 65°C. Incubate for 10 minutes at room temperature.
Centrifuge for 2 minutes at 14,000 to 18,000 g. If centrifuge has a "soft
setting," then use it.
Remove supernatant without disturbing Oligotex pellet. A little bit of
solution can be left behind to
reduce the loss of Oligotex. Save sup until certain that satisfactory binding
and elution of poly A+
mRNA has occurred.
Gently resuspend in Wash Buffer OW2 and pipet onto spin column. Centrifuge the
spin column
at full speed (soft setting if possible) for 1 minute.
Transfer spin column to a new collection tube and gently resuspend in Wash
Buffer OW2 and
centrifuge as describe herein.
Transfer spin column to a new tube and elute with 20 to 100 u1 of preheated
(70°C) Elution Buffer.
2 0 Gently resuspend Oligotex resin by pipetting up and down. Centrifuge as
above. Repeat elution
with fresh elution buffer or use first eluate to keep the elution volume low.
Read absorbance, using diluted Elution Buffer as the blank.
Before proceeding with cDNA synthesis, the mRNA must be precipitated.
Some component leftover or in the Elution Buffer from the Oligotex
purification procedure will
2 5 inhibit downstream enzymatic reactions of the mRNA.
Ethanol Precipitation
Add 0.4 vol. of 7.5 M NH40Ac + 2.5 vol. of cold 100% ethanol. Precipitate at -
20°C 1 hour to
overnight (or 20-30 min. at -70°C). Centrifuge at 14,000-16,000 x g for
30 minutes at 4°C. Wash
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pellet with 0.5m1 of 80%ethanol (-20°C) then centrifuge at 14,000-
16,000 x g for 5 minutes at room
temperature. Repeat 80% ethanol wash. Dry the last bit of ethanol from the
pellet in the hood.
(Do not speed vacuum). Suspend pellet in DEPC HZO at 1 ug/ul concentration.
Clean up total RNA using Qiagen's RNeasy kit
Add no more than 100ug to an RNeasy column. Adjust sample to.a volume of 100u1
with RNase-
free water. Add 350u1 Buffer RLT then 250u1 ethanol (100%) to the sample. Mix
by pipetting (do
not centrifuge) then apply sample to an RNeasy mini spin column. Centrifuge
for 15 sec at
>10,OOOrpm. If concerned about yield, re-apply flowthrough to column and
centrifuge again.
Transfer column to a new 2-ml collection tube. Add 500u1 Buffer RPE and
centrifuge for 15 sec
at >10,OOOrpm. Discard flowthrough. Add 500u1 Buffer RPE and centrifuge for 15
sec at
>10,OOOrpm. Discard flowthrough then centrifuge for 2 min at maximum speed to
dry column
membrane. Transfer column to a new 1.5-ml collection tube and apply 30-50u1 of
RNase-free
water directly onto column membrane. Centrifuge 1 min at >10,OOOrpm. Repeat
elution.
Take absorbance reading. If necessary, ethanol precipitate with ammonium
acetate and 2.5X
volume 100% ethanol.
Make cDNA using Gibco's "Superscript Choice System for cDNA Synthesis" kit
First Strand cDNA Synthesis
Use 5ug of total RNA or 1 ug of polyA+ mRNA as starting material. For total
RNA, use 2u1 of
Superscript RT. For polyA+ mRNA, use 1 u1 of Superscript RT. Final volume of
first strand
2 0 synthesis mix is 20u1. RNA must be in a volume no greater than 10u1.
Incubate RNA with 1 u1 of
100pmol T7-T24 oligo for 10 min at 70C. On ice, add 7 u1 of: 4u1 5X 15' Strand
Buffer, 2u1 of
0.1 M DTT, and 1 u1 of 10mM dNTP mix. Incubate at 37C for 2 min then add
Superscript RT
Incubate at 37C for 1 hour.
Second Strand Synthesis
2 5 Place 1 S' strand reactions on ice.
Add: 91 u1 DEPC H20
30u1 5X 2"d Strand Buffer
3u1 10mM dNTP mix
1 u1 10U/ul E.coli DNA Ligase
3 0 4u1 10U/ul E.coli DNA Polymerase
1 u1 2U/ul RNase H
Make the above into a mix if there are more than 2 samples. Mix and incubate 2
hours at 16C.
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Add 2u1 T4 DNA Polymerase. Incubate 5 min at 16C. Add 10u1 of 0.5M EDTA
Clean up cDNA
PhenoI:Chloroform:lsoamyl Alcohol (25:24:1 ) purification using Phase-Lock gel
tubes:
Centrifuge PLG tubes for 30 sec at maximum speed. Transfer cDNA mix to PLG
tube. Add equal
volume of phenol:chloroform:isamyl alcohol and shake vigorously (do not
vortex). Centrifuge 5
minutes at maximum speed. Transfer top aqueous solution to a new tube. Ethanol
precipitate:
add 7.5X 5M NH40ac and 2.5X volume of 100% ethanol. Centrifuge immediately at
room temp.
for 20 min, maximum speed. Remove sup then wash pellet 2X with cold 80%
ethanol. Remove
as much ethanol wash as possible then let pellet air dry. Resuspend pellet in
3u1 RNase-free
water.
In vitro Transcription (IVT) and labeling with biotin
Pipet 1.5u1 of cDNA into a thin-wall PCR tube.
Make NTP labeling mix:
Combine at room temperature: 2u1 T7 10xATP (75mM) (Ambion)
2u1 T7 10xGTP (75mM) (Ambion)
1.5u1 T7 10xCTP (75mM) (Ambion)
1.5u1 T7 10xUTP (75mM) (Ambion)
3.75u1 10mM Bio-11-UTP (Boehringer-Mannheim/Roche or
Enzo)
2 0 3.75u1 1 OmM Bio-16-CTP (Enzo)
2u1 10x T7 transcription buffer (Ambion)
2u1 10x T7 enzyme mix (Ambion)
Final volume of total reaction is 20u1. Incubate 6 hours at 37C in a PCR
machine.
RNeasy clean-up of IVT product
2 5 Follow previous instructions for RNeasy columns or refer to Qiagen's
RNeasy protocol handbook.
cRNA will most likely need to be ethanol precipitated. Resuspend in a volume
compatible with
the fragmentation step.
62
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15 ug of labeled RNA is usually fragmented. Try to minimize the fragmentation
reaction volume;
a 10 u1 volume is recommended but 20 u1 is all right. Do not go higher than 20
u1 because the
magnesium in the fragmentation buffer contributes to precipitation in the
hybridization buffer.
Fragment RNA by incubation at 94 C for 35 minutes in 1 x Fragmentation buffer.
5 x Fragmentation buffer:
200 mM Tris-acetate, pH 8.1
500 mM KOAc
150 mM MgOAc
The labeled RNA transcript can be analyzed before and after fragmentation.
Samples can be
heated to 65C for 15 minutes and electrophoresed on 1 % agarose/TBE gels to
get an
approximate idea of the transcript size range
Hybridization
200 u1 (10ug cRNA) of a hybridization mix is put on the chip. If multiple
hybridizations are to be
done (such as cycling through a 5 chip set), then it is recommended that an
initial hybridization
mix of 300 u1 or more be made.
Hybrization Mix: fragment labeled RNA (50ng/ul final cone)
50 pM 948-b control oligo
1.5 pM BioB
5 pM BioC
2 0 25 pM BioD
100 pM CRE
0.1mg/ml herring sperm DNA
0.5mg/ml acetylated BSA
to 300 u1 with 1xMES hyb. buffer
2 5 The instruction manuals for the products used herein are incorporated
herein in their entirety.
Labeling Protocol Provided Herein
Hybridization reaction:
Start with non-biotinylated IVT (purified by RNeasy columns)
(see example 1 for steps from tissue to IVT)
3 0 IVT antisense RNA; 4 fig: NI
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Random Hexamers (1 Ng/~tl): 4 ~I
HzO: ~I
14 ~I
- Incubate 70°C, 10 min. Put on ice.
Reverse transcription:
5X First Strand (BRL) 6 NI
buffer:
0.1 M DTT: 3 NI
50X dNTP mix: 0.6 NI
H20: 2.4 ~I
Cy3 or Cy5 dUTP (1 mM):3 NI
SS RT II (BRL): 1 NI
16 ~I
- Add to hybridization reaction.
- Incubate 30 min., 42°C.
- Add 1 NI SSII and let go for another hour.
Put on ice.
50X dNTP mix (25mM of cold dATP, dCTP, and dGTP, 10mM of dTTP: 25 ~I each of
100mM
2 0 dATP, dCTP, and dGTP; 10 ~I of 100mM dTTP to 15 ~I H20. dNTPs from
Pharmacia)
RNA degradation:
86 ~I HZO
- Add 1.5 ~I 1 M NaOH/ 2mM EDTA, incubate at 65°C, 10 min. 10 ~I 10N
NaOH
4 NI 50mM EDTA
2 5 U-Con 30
500 NI TE/sample spin at 70008 for 10 min, save flow through for purification
Qiagen purification:
-suspend u-con recovered material in 500N1 buffer PB
-proceed w/ normal Qiagen protocol
3 0 DNAse digest:
- Add 1 ~I of 1/100 dil of DNAse/30N1 Rx and incubate at 37°C for 15
min.
-5 min 95°C to denature enzyme
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Sample preparation:
- Add:
Cot-1 DNA: 10 NI
50X dNTPs: 1 ~I
20X SSC: 2.3 ~1
Na pyro phosphate: 7.5 NI
10mg/ml Herring sperm DNA 1u1 of 1/10 dilution
21.8 final vol.
- Dry down in speed vac.
- Resuspend in 15 ~I H20.
- Add 0.38 ~I 10% SDS.
- Heat 95°C, 2 min.
- Slow cool at room temp, for 20 min.
Put on slide and hybridize overnight at 64°C.
Washing after the hybridization:
3X SSC/0.03% SDS: 2 min. 37.5 mls 20X SSC+0.75m1s 10% SDS in 250m1s Hz0
1X SSC: 5 min. 12.5 mls 20X SSC in 250m1s H20
0.2X SSC: 5 min. 2.5 mls 20X SSC in 250m1s H20
Dry slides in centrifuge, 1000 RPM, 1 min.
2 0 Scan at appropiate PMT's and channels.
The results are shown in the tables and figures. The lists of genes come from
cells cultured in an
in vitro angiogenesis model. As indicated, some of the Accession numbers
include expression
sequence tags (ESTs). Thus, in one embodiment herein, genes within an
expression profile, also
termed expression profile genes, include ESTs and are not necessarily full
length.
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TABLE 1
Accession
Cluster#/ Gene Description
PROBESET
3 AB000450 vaccinia related kinase 2
4 AB002380 Human mRNA for KIAA382 gene; partial cds
4 AB003103 proteasome (prosome; macropain) 26S subunit;
non-ATPase; 12
4 AB004884 Homo sapiens mRNA for PKU-alpha; partial
cds
1 AF000573 rna1homogentisate 1;2-dioxygenase (homogentisate
oxidase)
3 AF008937 Homo sapiens syntaxin-16C mRNA, complete
cds
3 AF009301 Homo sapiens TEB4 protein mRNA; complete
cds
3 AF009368 Homo sapiens Luman mRNA; complete cds
4 D00591 chromosome condensation 1
4 D00760 proteasome (prosome; macropain) subunit;
alpha type; 2
tissue inhibitor of metalloproteinase 1
1 D11139 (erythroid potentiating activity;
collagenase inhibitor)
4 D14657 Human mRNA for KIAA11 gene; complete cds
4 D14878 D123 gene product
mannosyl (alpha-1;6-)-glycoprotein
1 D17716 beta-1;6-N-acetyl-glucosaminyltransferase
4 D21090 RAD23 (S. cerevisiae) homolog B
1 D26135 diacylglycerol kinase; gamma (9kD)
1 D26528 DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide
7 (RNA helicase; 52kD)
2 0 1 D30742 calcium/calmodulin-dependent protein kinase
IV
4 D31762 Human mRNA for KIAA57 gene; complete cds
4 D31765 Human mRNA for KIAA61 gene; partial cds
3 D31888 Homo sapiens clone 2479 mRNA sequence
4 D38128 prostaglandin I2 (prostacyclin) receptor
(1P)
2 5 2 D38500 postmeiotic segregation increased 2-like
4
4 D38551 RAD21 (S. pombe) homolog
4 D42087 Human mRNA for KIAA118 gene; partial cds
Human mRNA for Apo1_Human (MERS(Aop1-Mouse)-like
3 D49396 protein);
complete cds
Human monocyte PABL (pseudoautosomal boundary-like
4 D55640 sequence)
mRNA, clone Mo2
platelet-activating factor acetylhydrolase;
3 0 1 D63391 isoform Ib; gamma subunit
(29kD)
3 D63477 Human mRNA for KIAA143 gene; partial cds
4 D63483 acetyl LDL receptor; SREC
4 D64015 TIA1 cytotoxic granule-associated RNA-binding
protein-like 1
4 D79990 Human mRNA for KIAA168 gene; complete cds
3 5 4 D79997 Human mRNA for KIAA175 gene; complete cds
4 D80010 Human mRNA for KIAA188 gene; partial cds
1 D84276 CD38 antigen (p45)
4 D86425 Homo sapiens mRNA for nidogen-2
4 D86978 Human mRNA for KIAA225 ene; artial cds
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Accession
Cluster#/ Gene Description
PROBESET
1 D87012 Homo Sapiens clone 24675 mRNA sequence
4 D87075 Human mRNA for KIAA238 gene; partial cds
4 D87432 solute carrier family 7 (cationic amino
acid transporter; y+ system);
Homo sapiens mRNA for DNA topoisomerase
4 D87448 II binding protein; complete
cds
2 D87845 platelet-activating factor acetylhydrolase
2 (4kD)
1 HG1098-HT1098Cystatin D
4 HG2167-HT2237Protein Kinase Ht3l, Camp-Dependent
1 HG2415-HT2511Transcription Factor E2f-2
1 HG2825-HT2949Ret Transforming Gene
1 HG2887-HT3031Sry-Related Hmg-Box 12 Protein (Gb:X73039)
r
4 HG4660-HT5073Microtubule-Associated Protein 1 b
3 HG4704-HT5146Glial Growth Factor 2
4 HG884-HT884 Oncogene E6-Ap, Papillomavirus
1 HG919-HT919 Dna Polymerase, Epsilon, Catalytic Subunit
4 J00212 f Accession not listed in Genbank
4 J04029 keratin 1 (epidermolytic hyperkeratosis;
keratosis palmaris et plantaris)
5;1-methylenetetrahydrofolate dehydrogenase;
4 5;1-methylenetetrahydrofolate cyclohydrolase;
04031 1-formyltetrahydrofolate
synthetase
4 J04088 topoisomerase (DNA) II alpha (17kD)
4 J04543 annexin VII (synexin)
2 0 4 L06139 TEK tyrosine kinase; endothelial
1 L07540 ACTIVATOR 1 36 KD SUBUNIT
MADS box transcription enhancer factor
4 L08895 2; polypeptide C (myocyte
enhancer factor 2C)
1 L11239 gastrulation brain homeo box 1
1 L11353 neurofibromin 2 (bilateral acoustic neuroma)
4 L13773 Human AF-4 mRNA; complete cds
4 L13800 Homo Sapiens liver expressed protein gene,
3' end
4 L14922 replication factor C (activator 1 ) 1
(145kD)
4 L15189 heat shock 7kD protein 9B (mortalin-2)
4 L15388 Human G protein-coupled receptor kinase
(GRKS) mRNA, complete cds
3 0 3 L16895 lysyl oxidase
4 L27476 Friedreich ataxia region gene X14 (tight
junction protein ZO-2)
4 L27624 TISSUE FACTOR PATHWAY INHIBITOR 2 PRECURSOR
1 L32976 mixed lineage kinase 3
1 L33404 protease; serine; 6 (chymotryptic; stratum
corneum)
cytokine suppressive anti-inflammatory
3 5 4 L35263 drug binding protein 1 (p38 MAP
kinase)
1 L37347 natural resistance-associated macrophage
protein 2
4 L40371 thyroid hormone receptor interactor 4
4 L40391 Homo sa iens clone s153 mRNA fra ment
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Accession
Cluster#/ Gene Description
PROBESET
4 L41607 glucosaminyl (N-acetyl) transferase 2;
I-branching enzyme
1 L77566 Homo sapiens DGS-I mRNA; 3' end
1 M13928 aminolevulinate; delta-; dehydratase
1 M14016 uroporphyrinogen decarboxylase
4 M14219 decorin
4 M15796 proliferating cell nuclear antigen
4 M21305 Human alpha satellite and satellite 3 junction
DNA sequence
Human neural cell adhesion molecule (N-CAM)
4 M22092 gene, exon SEC and
partial cds
4 M22898 tumor protein p53 (Li-Fraumeni syndrome)
3 M22995 RAP1A; member of RAS oncogene family
3 M23379 RAS p21 protein activator (GTPase activating
protein) 1
1 M24364 major histocompatibility complex; class
II; DO beta 1
1 M24400 chymotrypsinogen B1
3 M25753 cyclin B1
4 M27691 cAMP responsive element binding protein
1
4 M28213 RAB2; member RAS oncogene family
SERINE/THREONINE PROTEIN PHOSPHATASE 2B
4 M29550 CATALYTIC
SUBUNIT; BETA ISOFORM
1 M29971 O-6-methylguanine-DNA methyltransferase
4 M30269 nidogen (enactin)
2 0 4 M31158 protein kinase; cAMP-dependent; regulatory;
type II; beta
3 M31166 pentaxin-related gene; rapidly induced
by IL-1 beta
3 M31210 endothelial differentiation; sphingolipid
G-protein-coupled receptor; 1
1 M55420 Human IgE chain, last 2 exons
prostaglandin-endoperoxide synthase 1 (prostaglandin
4 M59979 G/H synthase and
cyclooxygenase)
2 5 4 M62810 transcription factor 6-like 1 (mitochondria)
transcription factor 1-like)
4 M63838 interferon; gamma-inducible protein 16
1 M64710 Human C-type natriuretic peptide gene,
complete cds
Human phosphatidylcholine 2-acylhydrolase
3 M68874 (cPLA2) mRNA, complete
cds
3 M74524 ubiquitin-conjugating enzyme E2A (RAD6
homology
PEPTIDYL-PROLYL CIS-TRANS ISOMERASE; MITOCHONDRIAL
3 0 1 M80254 PRECURSOR
sphingomyelin phosphodiesterase 1; acid
1 M81780 cds3 lysosomal (acid
sphingomyelinase)
4 M83822 Human beige-like protein (BGL) mRNA; partial
cds
4 M86934 GS1 PROTEIN
1 M87338 replication factor C (activator 1 ) 2 (4kD)
3 5 1 M96326 rna1 azurocidin 1 (cationic antimicrobial protein
37)
4 M96954 TIA1 cytotoxic granule-associated RNA-binding
protein-like 1
4 M98833 Friend leukemia virus integration 1
1 S66793 arrestin 3; retinal X-arrestin
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Accession
Cluster#/ Gene Description
PROBESET
1 S72370 pyruvate carboxylase
4 S78569 laminin; alpha 4
4 S79873 lysosomal-associated membrane protein 2
1 S83325 aspartate beta-hydroxylase
putative RabS-interacting protein {clone
4 S83364 L1-57} [human, HeLa cells,
mRNA Partial, 366 nt)
putative RabS-interacting protein {clone
1 S83365 L1-94} [human, HeLa cells,
mRNA Partial, 369 nt)
1 001212 Human olfactory marker protein (OMP) gene,
complete cds
1 001922 deafness; X-linked 1; progressive
4 002556 Human RP3 mRNA; complete cds
4 002680 protein tyrosine kinase 9
4 003272 fibrillin 2
4 004209 Human associated microfibrillar protein
mRNA; complete cds
4 005237 fetal Alzheimer antigen
1 007225 purinergic receptor P2Y; G-protein coupled;
2
3 007620 protein kinase mitogen-activated 1 (MAP
kinase)
4 009759 protein kinase mitogen-activated 9 (MAP
kinase)
4 009820 alpha thalassemia/mental retardation syndrome
X-linked
3 011313 sterol carrier protein 2
3 014518 centromere protein A (17kD)
2 0 4 014575 protein phosphatase 1; regulatory (inhibitor)
subunit 8
3 015173 BCL2/adenovirus E1 B 19kD-interacting protein
2
4 015932 dual specificity phosphatase 5
4 018291 cell division cycle 16; anaphase promoting
complex 6
4 018300 damage-specific DNA binding protein 2 (48kD)
2 5 4 018383 nuclear respiratory factor 1
4 020536 caspase 6; apoptosis-related cysteine protease
4 021551 Human ECA39 mRNA; complete cds
4 023028 eukaryotic translation initiation factor
2B; subunit 5 (epsilon; 82kD)
1 023752 SRY (sex-determining region Y)-box 11
3 0 4 025435 Human transcriptional repressor (CTCF)
mRNA; complete cds
4 025997 stanniocalcin
4 028251 cds2 zinc finger protein 169
Human protein immuno-reactive with anti-PTH
4 028831 polyclonal antibodies
mRNA; partial cds
Human myelomonocytic specific protein (MNDA)
4 030245 gene, 5' flanking
sequence and complete exon 1
3 5 4 032315 Human syntaxin 3 mRNA; complete cds
4 032439 regulator of G-protein signalling 7
3 032849 N-myc (and STAT) interactor
4 035139 necdin (mouse) homolog
1 036764 eukaryotic translation initiation factor
3; subunit 2 (beta; 36kD)
4 0 4 039400 chromosome 11 open reading frame 4
~ U39657 protein kinase; mitogen-activated; kinase
6 (MAP kinase kinase 6)
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Accession
Cluster#/ Gene Description
PROBESET
4 041344 proline arginine-rich end leucine-rich
repeat protein
3 041766 a disintegrin and metalloproteinase domain
9 (meltrin gamma)
3 041813 homeo box A9
3 041815 Human nucleoporin 98 (NUP98) mRNA, complete
cds
4 043286 Human selenophosphate synthetase 2 (SPS2)
mRNA; complete cds
4 044378 MAD (mothers against decapentaplegic; Drosophila)
homolog 4
4 044754 small nuclear RNA activating complex; polypeptide
1; 43kD
1 047011 cdsl fibroblast growth factor 8 (androgen-induced)
Human DNA-dependent protein kinase catalytic
4 047077 subunit (DNA-PKcs)
mRNA; complete cds
4 048251 Homo sapiens protein kinase C-binding protein
RACK7 mRNA; partial cds
4 050535 Human BRCA2 region; mRNA sequence CG6
4 056833 von Hippel-Lindau binding protein 1
4 058091 cullin 4B
1 058837 cyclic nucleotide gated channel beta 1
4 059289 cadherin 13; H-cadherin (heart)
4 059863 TNF receptor-associated factor 2
4 067122 ubiquitin-like 1 (sentrin)
4 067319 caspase 7; apoptosis-related cysteine protease
3 068019 MAD (mothers against decapentaplegic; Drosophila)
homolog 3
a disintegrin and metalloproteinase domain
2 0 1 069611 17 (tumor necrosis factor;
alpha; converting enzyme)
4 070322 karyopherin (importin) beta 2
4 073524 Human putative ATP/GTP-binding protein
(HEAB) mRNA; complete cds
4 079267 Human clone 2384 mRNA; partial cds
4 079291 Human clone 23721 mRNA sequence
2 5 4 082671 cds2 Homo sapiens clone LM1955 H15e3 gene; partial
cds
4 084573 procollagen-lysine; 2-oxoglutarate 5-dioxygenase
(lysine hydroxylase) 2
4 090914 carboxypeptidase D
1 091316 Homo Sapiens mRNA for brain acyl-CoA hydrolase;
complete cds
4 091932 clathrin-associated/assemblyladaptor protein;
small 3; 22-kD; Sigma3A
3 0 4 096131 Homo sapiens HPV16 E1 protein binding protein
mRNA; complete cds
4 097018 echinoderm microtubule-associated protein-like
Homo Sapiens putative RNA binding protein
4 097188 KOC (koc) mRNA; complete
cds
4 V00503 collagen; type I; alpha 2
3 X04327 2;3-bisphosphoglycerate mutase
3 5 1 X06389 synaptophysin
1 X07496 apolipoprotein A-I
2 X07820 matrix metalloproteinase 1 (stromelysin
2)
3 X14787 thrombos ondin 1
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Accession
Cluster#/ Gene Description
PROBESET
4 X15525 rna1 acid phosphatase 2; lysosomal
NAD-DEPENDENT METHYLENETETRAHYDROFOLATE
3 X16396 DEHYDROGENASE
4 X16609 ankyrin 1; erythrocytic
4 X53586 rnal Human mRNA for integrin alpha 6
4 X53793 MULTIFUNCTIONAL PROTEIN ADE2
1 X54936 placental growth factor; vascular endothelial
growth factor-related protein
4 X55740 5' nucleotidase (CD73)
2 X57025 insulin-like growth factor 1 (somatomedin
C)
2 X60673 rna1 adenylate kinase 3
dipeptidylpeptidase IV (CD26; adenosine
4 X60708 deaminase complexing protein
2)
4 X62048 wee1+ (S, pombe) homolog
2 X63097 Rhesus blood group; D antigen
4 X63563 polymerase (RNA) II (DNA directed) polypeptide
B (14kD)
4 X64037 general transcription factor IIF; polypeptide
1 (74kD subunit)
4 X69636 hect domain and RLD 2
4 X69878 fms-related tyrosine kinase 4
4 X70649 DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide
1
3 X72841 H.sapiens IEF 7442 mRNA
4 X74987 ribonuclease L (2 ;5'-oligoisoadenylate
synthetase-dependent) inhibitor
2 0 4 X83107 BMX non-receptor tyrosine kinase
3 X84194 acylphosphatase 1; erythrocyte (common)
type
4 X85753 cyclin-dependent kinase 8
1 X87870 H.sapiens mRNA for hepatocyte nuclear factor
4a
4 X89066 transient receptor potential channel 1
2 5 4 X89398 cds2 uracil-DNA glycosylase
1 X89399 Homo sapiens mRNA for Ins(1;3;4;5)P4-binding
protein
3 X89426 H.sapiens mRNA for ESM-1 protein
4 X91247 thioredoxin reductase 1
4 X91648 H.sapiens mRNA for pur alpha extended 3'untranslated
region
3 0 4 X92098 H.sapiens mRNA for transmembrane protein
rnp24
4 X92110 H.sapiens mRNA for hcgVlll protein
4 X94703 RAB28; member RAS oncogene family
1 X96506 H.sapiens mRNA for NC2 alpha subunit
Homo sapiens natural killer-associated
1 X97230 f transcript 5 (NKAT5) mRNA;
complete cds
3 5 4 X98263 H.sapiens mRNA for M-phase phosphoprotein;
mpp6
ubiquitin specific protease 9; X chromosome
4 X98296 (Drosophila fat facets
related)
4 X99584 H.sa lens mRNA for SMT3A rotein
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Accession
Cluster#/ Gene Description
PROBESET
4 Y00264 amyloid beta (A4) precursor protein (protease
nexin-II; Alzheimer disease)
4 Y07566 H.sapiens mRNA for RIT protein
3 Y07759 myosin VA (heavy polypeptide 12; myoxin)
1 Y07827 Human butyrophilin (BTFS) mRNA; complete
cds
4 Y07867 pirin
4 Y09443 alkylglycerone phosphate synthase
4 Y09858 H.sapiens mRNA for unknown protein
4 Y12394 karyopherin alpha 3 (importin alpha 4)
3 211559 iron-responsive element binding protein
1
4 211695 protein kinase; mitogen-activated 1 (MAP
kinase 1; p4; p41)
3 215005 centromere protein E (312kD)
1 246261 H.sapiens DNA for histone H3a
2 AA011243 s ESTs
2 AA018418 ESTs
2 AA018758 ESTs
2 AA018804 Homo sapiens clone 23675 mRNA sequence
3 AA031993 Homo sapiens HRIHFB2115 mRNA; partial cds
2 AA044217 ESTs; Weakly similar to similar to cuticle
collagen [C.elegans]
SWI/SNF related; matrix associated; actin
4 AA046548 dependent regulator of
chromatin; subfamily e; member 1
ESTs; Moderately similar to !!!! ALU SUBFAMILY
2 0 2 AA057447 s SB WARNING ENTRY
!!!! [H.sapiens]
Sjogren syndrome antigen A2 (6kD; ribonucleoprotein
2 AA058376 autoantigen
SS-A/Ro)
4 AA083572 v-ral simian leukemia viral oncogene homolog
A (ras related)
4 AA085696 ESTs
2 AA088744 ESTs
ESTs; Weakly similar to putative T1/ST2
2 5 2 AA089688 receptor binding protein
precursor [H.sapiens]
4 AA091284 ESTs
2 AA092700 ESTs
1 AA092968 ESTs
4 AA094800 eukaryotic translation initiation factor
3; subunit 7 (zeta; 66/67kD)
3 0 4 AA100219 ESTs
4 AA114885 ESTs
ESTs; Weakly similar to !!!! ALU SUBFAMILY
4 AA129547 SQ WARNING ENTRY !!!!
[H.sapiens]
4 AA133016 ESTs
3 AA149507 homolog of mouse quaking QKI (KH domain
RNA binding protein)
3 5 2 AA151005 sperm surface protein
zp61b6.r1 Stratagene endothelial cell 937223
4 AA187101 Homo sapiens cDNA clone
IMAGE:624659 5', mRNA sequence
3 AA195179 s ESTs
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Accession
Cluster#/ Gene Description
PROBESET
2 AA203138 low density lipoprotein receptor (familial
hypercholesterolemia)
2 AA203645 ESTs; Moderately similar to SH3-containing
protein p415 [R.norvegicus]
zq54c6.r1 Stratagene neuroepithelium (#937231
3 ) Homo sapiens cDNA
A206236 clone IMAGE:645418 5' similar to TR:G122922
6122922 ALLOGRAFT
INFLAMMATORY FACTOR-1. ;, mRNA sequence
1 AA227621 ESTs; Weakly similar to weak similarity
to collagens [C.elegans]
ESTs; Weakly similar to X-linked retinopathy
4 AA248283 protein {C-terminal; clone
XEH.Bc} [H.sapiens]
3 AA249611 H.sapiens mRNA for 21-Glutamic Acid-Rich
Protein (21-GARP)
2 AA282640 ESTs
2 AA287199 Human mRNA for KIAA81 gene; partial cds
ESTs; Highly similar to HYPOTHETICAL 3.5
2 AA313990 KD PROTEIN C3A5.3 IN
CHROMOSOME III [Caenorhabditis elegans]
EST18611 Colon carcinoma (HCC) cell line
2 AA314256 II Homo sapiens cDNA 5' end,
mRNA sequence
ESTs; Highly similar to ADP-RIBOSYLATION
2 AA314389 FACTOR 1
[Saccharomyces cerevisiae]
ESTs; Moderately similar to !!!! ALU SUBFAMILY
2 AA324364 J WARNING ENTRY !!!!
[H.sapiens]
3 AA329211 s ESTs
2 AA399187 ESTs
4 AA421079 ESTs
2 AA422029 ESTs
3 AA425230 Human GAP SH3 binding protein mRNA; complete
cds
ESTs; Highly similar to N-terminal asparagine
4 AA447052 amidohydrolase
[M.musculus]
4 AA452000 ESTs
2 0 4 AA456687 ESTs
ESTs; Weakly similar to X-linked retinopathy
4 AA487015 s protein {C-terminal; clone
XEH.Bc} [H.sapiens]
2 AB002326 Human mRNA for KIAA328 gene; partial cds
4 AFFX-BioB-3
2 C01527 ESTs
2 5 4 C01714 Homo sapiens serum-inducible kinase mRNA;
complete cds
3 C01811 f ESTs
ESTs; Moderately similar to !!!! ALU SUBFAMILY
2 C02352 s SO WARNING ENTRY
!!!! [H.sapiens]
1 C02375 Human mRNA containing an Alu repeat and
its reverse complement
2 C14448 EST
3 0 4 D16611 s coproporphyrinogen oxidase (coproporphyria;
harderoporphyria)
2 D25216 Human mRNA for KIAA14 gene; complete cds
2 D31352 ESTs; Weakl similar to h othetical rotein
H.sa lens
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Accession
Cluster#/ Gene Description
PROBESET
ESTs; Weakly similar to probable hormone
4 D58024 s receptor EMR1 precursor
[H.sapiens]
1 D80897 Homo sapiens clone 24736 mRNA sequence
3 D82614 ESTs
4 D87845 platelet-activating factor acetylhydrolase
2 (4kD)
1 D89377 i msh (Drosophila) homeo box homolog 2
2 H06583 cAMP responsive element binding protein-like
2
1 H40732 ESTs
yp19h1.r1 Soares breast 3NbHBst Homo sapiens
4 H46617 cDNA clone
IMAGE:187921 5', mRNA sequence
1 H56731 ESTs
1 0 1 H75570 ESTs
2 H78886 ESTs
ESTs; Highly similar to ERYTHROID KRUEPPEL-LIKE
1 H81241 TRANSCRIPTION
FACTOR [Mus musculus]
1 L36531 integrin; alpha 8
2 M63154 gastric intrinsic factor (vitamin B synthesis)
4 M63180 threonyl-tRNA synthetase
2 M91504 ESTs
2 N56191 Homo sapiens protocadherin 68 (PCH68) mRNA;
complete cds
2 N78483 ESTs
2 N79268 zinc finger protein 198
2 0 2 814652 Homo sapiens PAC clone DJ872F7 from 7q31
yg33f12.r1 Soares infant brain 1NIB Homo
2 820459 sapiens cDNA clone
IMAGE:34345 5', mRNA sequence
3 822303 ESTs; Weakly similar to putative p15 [H.sapiens]
2 833779 ESTs; Weakly similar to p4 [H.sapiens]
2 836553 ESTs; Weakly similar to KIAA681 protein
[H.sapiens]
2 5 2 864534 ESTs
4 866475 ESTs
4 870621 Homo sapiens mRNA for KIAA896 protein;
partial cds
3 879356 ESTs; Weakly similar to PROTEIN Q3 [Mus
musculus]
2 884933 ESTs; Weakly similar to putative p15 [H.sapiens]
3 0 3 RC AA007160 ESTs
ESTs; Highly similar to protein tyrosine
2 RC AA007234 phosphatase epsilon cytoplasmic
s isoform [H.sapiens]
2 RC AA018409 ESTs
4 RC AA025351 ESTs
3 RC AA027168 ESTs
ESTs; Weakly similar to !!!! ALU SUBFAMILY
3 5 1 RC AA027317 J WARNING ENTRY !!!!
[H.sapiens]
3 RC AA029423 ESTs
4 RC AA031357 ESTs
4 RC AA045136 ESTs
1 RC AA053400 ESTs
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Accession
Cluster#/ Gene Description
PROBESET
ESTs; Weakly similar to !!!! ALU SUBFAMILY
3 RC AA055829 J WARNING ENTRY !III
[H.sapiens]
3 RC AA065217 ESTs
1 RC AA116054 ESTs
1 RC AA126311 ESTs
4 RC AA129390 ESTs
ESTs; Highly similar to DOSAGE COMPENSATION
4 RC AA130273 REGULATOR
[Drosophila melanogaster]
2 RC AA142919 ESTs
4 RC AA150205 ubiquitous Kruppel-like transcription factor
1 RC AA176867 ESTs
ESTs; Highly similar to U1 small nuclear
2 RC AA180321 ribonucleoprotein 1 SNRP
homolog [H.sapiens]
2 RC_AA180487 Homo sapiens TACC1 (TACC1 ) mRNA; complete
cds
4 RC AA187634 eukaryotic translation initiation factor
3; subunit 1 (alpha; 35kD)
3 RC AA195399 ESTs
3 RC AA234717 ESTs
4 RC AA234743 ESTs
3 RC AA234957 Homo sapiens mRNA for MTMR1 protein
3 RC AA235604 ESTs
ESTs; Weakly similar to PROBABLE E5 PROTEIN
3 RC AA236559 [Human papillomavirus
type 58]
3 RC AA242868 ESTs; Weakly similar to house-keeping protein
[M.musculus]
2 0 4 RC AA251776 jun D proto-oncogene
4 RC AA251909 Homo sapiens protein kinase homolog (BUBR1
) mRNA; complete cds
diptheria toxin resistance protein required
3 RC AA252672 for diphthamide biosynthesis
s (Saccharomyces)-like 2
3 RC AA256157 ESTs
4 RC AA256680 ESTs
2 5 3 RC AA258873 ESTs
1 RC AA262727 ESTs
4 RC AA281451 ESTs
4 RC AA281545 ESTs
3 RC AA282069 Homo sapiens mRNA for KIAA63 protein; complete
cds
3 0 1 RC AA283044 ESTs
3 RC AA283930 ESTs
4 RC AA284755 ESTs; Weakly similar to unknown [H.sapiens]
4 RC AA291268 ESTs
1 RC AA291927 ESTs
3 5 2 RC AA343514 ESTs
3 RC AA398109 ESTs
4 RC AA405737 ESTs
4 RC AA406610 ESTs
4 RC AA411465 ESTs
4 0 3 RC AA416886 ESTs; Weakl similar to redicted usin Genefinder
C.ele ans
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Accession
Cluster#/ Gene Description
PROBESET
4 RC AA424013 Homo sapiens clone 23767 and 23782 mRNA
sequences
4 RC_AA424148 ESTs
2 RC_AA424558 ESTs; Weakly similar to 33-kDa phototransducing
protein [H.sapiens]
4 RC AA424961 Homo sapiens TEB4 protein mRNA; complete
s cds
3 RC_AA425367 ESTs
1 RC_AA425921 Homo sapiens I-1 receptor candidate protein
mRNA; complete cds
4 RC AA426220 Homo sapiens mRNA for KIAA523 protein;
partial cds
4 RC AA427735 ESTs
4 RC AA430673 ESTs
4 RC AA432248 ESTs
4 RC_AA435896 ESTs
3 RC_AA436705 Homo sapiens mRNA for KIAA766 protein;
complete cds
3 RC_AA446561 Homo sapiens mRNA for KIAA47 protein; complete
cds
4 RC_AA448238 Homo Sapiens mRNA for KIAA915 protein;
complete cds
3 RC AA448688 ESTs; Weakly similar to KIAA638 protein
[H.sapiens)
3 RC AA449756 ESTs; Weakly similar to rA8 [R.norvegicus)
4 RC AA450303 ESTs
3 RC AA452411 ESTs
4 RC AA454566 ribosomal protein L13
2 0 4 RC AA454667 ESTs
4 RC AA456437 ESTs
4 RC AA456646 ESTs
4 RC AA456826 ESTs
4 RC AA456981 ESTs
2 5 4 RC AA458959 ESTs
3 RC AA459950 ESTs
ESTs; Highly similar to PROBABLE PHOSPHOSERINE
3 RC AA460449 AMINOTRANSFERASE [Oryctolagus cuniculus]
3 RC AA463910 ESTs
4 RC AA464603 ESTs
3 0 3 RC AA464606 ESTs
4 RC AA465093 TIA1 cytotoxic granule-associated RNA-binding
protein
3 RC AA465692 Homo sapiens mRNA for KIAA648 protein;
partial cds
3 RC AA476473 Homo sapiens Trio mRNA; complete cds
1 RC AA478109 ESTs
3 5 4 RC AA478474 ESTs
3 RC AA480889 ESTs
1 RC AA485223 ESTs
1 RC AA485254 ESTs
4 RC AA486183 ESTs; Weakly similar to Yhr1wp [S.cerevisiae]
4 0 3 RC AA496936 ESTs; Weakly similar to B cell growth factor
[H.sapiens]
4 RC AA598589 ESTs
4 RC AA598831 ESTs
f
4 RC AA600150 ESTs
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Accession
Cluster#/ Gene Description
PROBESET
4 RC AA608545 ESTs
3 RC AA609210 ESTs
ESTs; Highly similar to PROBABLE PEPTIDYL-PROLYL
3 RC_AA610108 CIS-TRANS
ISOMERASE C21E11.5C [Schizosaccharomyces
pombe]
4 RC AA620582 ESTs; Weakly similar to (defline not available
424227) [H.sapiens]
ESTs; Highly similar to HYPOTHETICAL 98.3
4 RC AA621239 KD PROTEIN R1 E12.1 IN
CHROMOSOME III [Caenorhabditis elegans]
3 RC AA621714 ESTs
1 RC AA621718 ESTs
1 RC D19673 ESTs
1 RC D25755 ESTs
s
1 RC_D51095 ESTs
4 RC D60272 ESTs; Weakly similar to macrophage lectin
i 2 [H.sapiens]
2 T08879 cathepsin F
UDP-N-acetyl-alpha-D-galactosamine:polypeptide
3 T34527 N-acetylgalactosaminyltransferase 1 (GaINAc-T1
)
2 T40327 s ESTs
3 T62771 s Homo sapiens nucleoplasmin-3 (NPM3) mRNA;
complete cds
1 T63174 s ESTs; Weakly similar to neuronal thread
protein AD7c-NTP [H.sapiens]
2 T83444 Homo sapiens mRNA for KIAA887 protein;
partial cds
1 T93641 ESTs
2 048263 prepronociceptin
2 0 2 049065 interleukin 1 receptor-like 2
2 079300 Human clone 23629 mRNA sequence
1 088573 Human NBR2 mRNA; complete cds
2 093867 Human RNA polymerase III subunit (RPC62)
mRNA; complete cds
4 W01094 ESTs
2 5 2 W01568 ESTs
2 W26853 ESTs
2 W27179 BCL2/adenovirus E1B l9kD-interacting protein
3-like
2 W27965 epimorphin
Homo sapiens RRM RNA binding protein Gry-rbp
3 W36280 s (GRY-RBP) mRNA;
complete cds
3 0 2 W47063 ESTs
4 W79060 ESTs; Weakly similar to Ras-binding protein
SUR-8 [M.musculus]
4 W88550 ESTs; Moderately similar to trg gene product
[R.norvegicus]
1 X60486 H4 histone family; member G
2 X78931 s H.sapiens HZF8 mRNA for zinc finger protein
3 5 1 214077 s YY1 transcription factor
1 RC AA002147 EST
1 RC AA004711 ESTs
1 RC AA010383 EST
1 RC AA015761 ESTs
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Accession
Cluster#/ Gene Description
PROBESET
2 RC AA018772 ESTs
2 RC AA021473 EST
r
potassium voltage-gated channel; delayed-rectifier;
2 RC AA024835 subfamily S; member
3
2 RC AA025858 ESTs
1 RC AA027229 ESTs
1 RC AA029428 ESTs
3 RC AA035143 ESTs
1 RC AA035237 ESTs
2 RC AA039347 EST
1 RC AA040740 ESTs
3 RC AA041551 ESTs
1 RC AA045513 ESTs
1 RC AA045745 EST
1 RC AA055348 ESTs
2 RC AA056582 ESTs
s
1 RC AA056697 ESTs
1 RC AA056746 EST
3 RC AA057678 ESTs
2 RC AA058681 ESTs
2 0 2 RC AA058686 ESTs
zm5c1.s1 Stratagene corneal stroma (#937222)
2 Homo sapiens cDNA
C AA062840 clone IMAGE:513234 3' similar to gb:S71381
PROTEASOME BETA
CHAIN (HUMAN);, mRNA sequence
zm5f3.s1 Stratagene fibroblast (#937212)
2 RC AA064859 Homo sapiens cDNA clone
IMAGE:52985 3', mRNA sequence
zm12e11.s1 Stratagene pancreas (#93728)
1 RC AA065069 Homo Sapiens cDNA clone
IMAGE:525452 3', mRNA sequence
zm67g3.s1 Stratagene neuroepithelium (#937231
) Homo Sapiens cDNA
1 clone IMAGE:5374 3' similar to gb:S66915_cds1
C AA069923 ATP SYNTHASE
GAMMA CHAIN, MITOCHONDRIAL PRECURSOR (HUMAN);,
mRNA
sequence
zm6h5.s1 Stratagene fibroblast (#937212)
2 RC AA070799 Homo Sapiens cDNA clone
s IMAGE:5373 3', mRNA sequence
zm6a1.s1 Stratagene fibroblast (#937212)
2 Homo Sapiens cDNA clone
C AA070815 IMAGE:529992 3' similar to gb:X67951 PROLIFERATION-ASSOCIATED
PROTEIN PAG (HUMAN);, mRNA sequence
zm87a1.s1 Stratagene ovarian cancer (#937219)
2 RC AA075374 Homo sapiens cDNA
clone IMAGE:544872 3', mRNA sequence
zm91g8.s1 Stratagene ovarian cancer (#937219)
2 RC AA076382 Homo Sapiens cDNA
clone IMAGE:545342 3', mRNA sequence
1 RC AA078787 ESTs
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Accession
Cluster#/ Gene Description
PROBESET
zm92h1.s1 Stratagene ovarian cancer (#937219)
2 RC AA078986 Homo Sapiens cDNA
clone IMAGE:545425 3', mRNA sequence
zm95h11.s1 Stratagene colon HT29 (#937221
1 ) Homo Sapiens cDNA
C AA079393 clone IMAGE:545733 3' similar to gb:X1656
CYTOCHROME C OXIDASE
POLYPEPTIDE VIIC PRECURSOR (HUMAN);, mRNA
sequence
zm97f8.s1 Stratagene colon HT29 (#937221
2 RC AA079487 ) Homo sapiens cDNA clone
IMAGE:545895 3', mRNA sequence
2 RC AA083207 EST
2 RC AA083256 vinculin
zn6g9.s1 Stratagene hNT neuron (#937233)
2 RC AA084415 Homo Sapiens cDNA clone
IMAGE:546688 3', mRNA sequence
znlf1.s1 Stratagene colon HT29 (#937221
2 ) Homo sapiens cDNA clone
C AA085274 IMAGE:546169 3' similar to gb:X15341 CYTOCHROME
C OXIDASE
POLYPEPTIDE VIA-LIVER (HUMAN);, mRNA sequence
2 RC_AA088678 ESTs
3 RC AA100925 ESTs; Weakly similar to predicted using
Genefinder [C.elegans]
ESTs; Highly similar to J KAPPA-RECOMBINATION
3 RC AA101255 SIGNAL BINDING
PROTEIN [Homo sapiens]
3 RC AA126474 stanniocalcin 2
2 RC AA127017 ESTs
ESTs; Weakly similar to protein phosphatase
2 RC AA129968 2A 13 kDa regulatory
subunit [H.sapiens]
2 RC AA130240 ESTs
1 RC AA131866 ESTs
ESTs; Moderately similar to !!!! ALU SUBFAMILY
2 RC AA132039 J WARNING ENTRY !!!!
[H.sapiens)
ESTs; Moderately similar to C-1-TETRAHYDROFOLATE
3 RC AA132983 SYNTHASE;
CYTOPLASMIC [Saccharomyces cerevisiae]
ESTs; Weakly similar to NADH-UBIQUINONE
3 RC AA133250 OXIDOREDUCTASE
CHAIN 4 [Caenorhabditis elegans]
1 RC AA133583 high-mobility group (nonhistone chromosomal)
s protein isoform I-C
2 0 4 RC AA135941 ESTs
zo9e6.s1 Stratagene neuroepithelium NT2RAM1
2 RC AA148650 937234 Homo Sapiens
cDNA clone IMAGE:56722 3', mRNA sequence
2 RC AA151110 ESTs
ESTs; Moderately similar to !!!! ALU SUBFAMILY
2 RC AA155754 SX WARNING ENTRY
!!!! [H.sapiens]
4 RC AA156125 ESTs
2 5 2 RC AA156289 ESTs
1 RC AA156997 ESTs
2 RC AA157291 ESTs
CA 02381699 2002-02-07
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Accession
Cluster#/ Gene Description
PROBESET
2 RC AA157293 ESTs
2 RC AA164293 ESTs
f
1 RC_AA164676 EST
1 RC AA167375 Homo sapiens mRNA for KIAA53 protein; partial
cds
ESTs; Moderately similar to !!!! ALU SUBFAMILY
1 RC AA167550 SX WARNING ENTRY
!!!! [H.sapiens]
2 RC AA176589 EST
1 RC AA180448 EST
4 RC AA187144 endothelin 1
s
3 RC AA189170 ESTs
f
4 RC AA192757 ESTs
2 RC AA205650 ESTs
ESTs; Weakly similar to neural differentiation-associated
4 RC AA233342 protein
[M.musculus]
3 RC AA233472 ESTs
2 RC AA234110 ESTs
4 RC D80981 ESTs
ESTs; Weakly similar to HYPOTHETICAL PROTEIN
3 RC F01660 H134 [Haemophilus
influenzae]
1 RC F02206 EST; Highly similar to ether-a-go-go-related
protein [H.sapiens]
4 RC F02208 ESTs
4 RC F02544 ESTs
2 0 4 RC F03918 ESTs
4 RC F04258 ESTs; Highly similar to INORGANIC PYROPHOSPHATASE
s [Bos taurus]
4 RC F04600 ESTs
4 RC F08998 ESTs
2 RC F09605 ESTs
4 RC F11115 ESTs
3 RC H06371 ESTs
1 RC H10995 ESTs
ESTs; Weakly similar to HYPOTHETICAL 97.6
1 RC H11938 KD PROTEIN IN
SHP1-SEC17 INTERGENIC REGION [Saccharomyces
cerevisiae]
4 RC H16568 ESTs
3 0 4 RC H 16772 ESTs
ESTs; Moderately similar to seven-pass
1 RC H18951 transmembrane receptor
precursor [M.musculus]
1 RC H20859 ESTs
1 RC H23747 ESTs
1 RC H38087 ESTs
3 5 1 RC H40331 ESTs
1 RC H40567 ESTs
81
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Accession
Cluster#/ Gene Description
PROBESET
1 RC H46966 ESTs
yq99a5.s1 Soares fetal liver spleen 1 NFLS
1 RC_H56640_i Homo Sapiens cDNA clone
I MAGE:23888 3', mRNA sequence
1 RC H57154 ESTs; Weakly similar to RST [M.musculus]
1 RC H96712 ESTs
1 RC_N20814 ESTs
3 RC N25249 synaptosomal-associated protein; 23kD
1 RC_N27100 ESTs
1 RC N39616 RNA (guanine-7-) methyltransferase
1 RC N48982 ESTs
1 RC_N51957 ESTs
1 RC_N52271 Homo Sapiens LIM protein mRNA; complete
cds
1 RC_N59435 ESTs; Weakly similar to No definition line
found [H.sapiens]
1 RC N64139 ESTs; Weakly similar to Ndr protein kinase
[H.sapiens]
3 RC N66981 ESTs
1 RC N68640 ESTs
ESTs; Highly similar to PRE-MRNA SPLICING
4 RC_N69352 FACTOR RNA HELICASE
PRP22 [Saccharomyces cerevisiae]
4 RC N95226 Homo sapiens mRNA for KIAA758 protein;
partial cds
1 RC 800138 ESTs
ESTs; Weakly similar to !!!! ALU SUBFAMILY
1 RC_R07998 J WARNING ENTRY !!!!
[H.sapiens]
2 0 1 RC 808929 ubiquitin-conjugating enzyme E2G 2 (homologous
to yeast UBC7)
1 RC 810307 ESTs
3 RC 833354 ESTs
1 RC 836083 ESTs
1 RC 837938 ESTs
f
ydlg4.s1 Soares infant brain 1NIB Homo
2 5 1 RC 839330 Sapiens cDNA clone
IMAGE:24282 3', mRNA sequence
1 RC 840816 cullin 4A
s
1 RC 843162 ESTs
s
ESTs; Weakly similar to Similarity to Salmonella
3 RC 845698 regulatory protein UHPC
(C.elegans]
2 RC 854554 ESTs
ESTs; Weakly similar to alternatively spliced
3 0 1 RC 868425 product using exon 13A
[H.sapiens]
1 RC 868568 ESTs
3 RC 868763 ESTs
1 RC 870467 ESTs
ESTs; Moderately similar to !!!! ALU SUBFAMILY
1 RC_R73565 SX WARNING ENTRY
!!!! [H.sapiens]
3 5 4 RC 873640 ESTs
82
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Accession
Cluster#/ Gene Description
PROBESET
1 RC 878376 EST
1 RC 892453 EST
1 RC T03865 ESTs
3 RC T03872 ESTs
1 RC T10072 ESTs
1 RC_T10080 ESTs
1 RC T10132 Homo sapiens mRNA for KIAA478 protein;
complete cds
1 RC T15343 ESTs
2 RC T23457 ESTs
1 RC T23555 ESTs
2 RC T23670 ESTs
4 RC T23948 ESTs
4 RC T33464 ESTs
1 RC T34413 ESTs
2 RC T34611 ESTs
2 RC T40920 ESTs
4 RC T55182 ESTs
2 RC T77453 EST
1 RC T84039 ESTs
2 0 1 RC T86458 ESTs
1 RC T87693 ESTs
2 RC T89350 ESTs
s
1 RC T90945 ESTs
2 RC T90987 ESTs
2 5 1 RC T91863 ESTs
1 RC T91881 EST
1 RC T93783 ESTs
s
1 RC T96687 ESTs
2 RC T96944 ESTs
3 0 3 RC T97307 ESTs; Weakly similar to neuronal thread
protein AD7c-NTP [H.sapiens]
1 RC T97764 ESTs
2 RC W48817 ESTs
2 RC W58343 ESTs
1 RC W59949 ESTs; Highly similar to RAS-LIKE PROTEIN
TC1 [Homo sapiens]
3 5 1 RC W74644 ESTs
ESTs; Highly similar to UBIQUITIN-CONJUGATING
1 RC W74761 ENZYME E2-17 KD
[Caenorhabditis elegans]
1 RC W74802 ESTs
1 RC W81205 ESTs
2 RC W81237 ESTs
4 0 3 RC W90146 ESTs
f
1 RC W92798 ESTs
1 RC 238412 EST
1 RC 238709 inositol 1;4;5-tri hos hate rece tor; t
a 2
83
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Accession
Cluster#/ Gene Description
PROBESET
1 RC_Z38904 ESTs
2 RC 239103 core-binding factor; runt domain; alpha
subunit 2; translocated to; 2
2 RC 239930 ESTs
f
2 RC_Z39939 ESTs
3 RC 240012 Homo sapiens mRNA for KIAA587 protein;
i complete cds
2 RC 240377 ESTs
s
1 RC 240820 ESTs
3 RC 241680 ESTs
4 AFFX-BioB-3
2 RC AA005112 Human zinc-finger domain-containing protein
mRNA; partial cds
ESTs; Highly similar to ANTI-SILENCING
4 RC AA005432 PROTEIN 1 [Saccharomyces
cerevisiae]
4 RC AA010163 Human mRNA for KIAA312 gene; partial cds
4 RC AA026356 ESTs
2 RC AA026901 ESTs
4 RC AA036867 ESTs
1 RC_AA044644 Pp52
4 RC AA046426 Homo Sapiens MSE55-related protein (UB1
) mRNA; complete cds
ESTs; Weakly similar to X-linked retinopathy
4 RC AA054515 protein {C-terminal; clone
XEH.Bc} [H.sapiens]
zn17h6.s1 Stratagene neuroepithelium NT2RAM1
2 RC AA084162 937234 Homo Sapiens
cDNA clone IMAGE:547739 3', mRNA sequence
2 0 4 RC AA085749 Homo sapiens mRNA for ATP binding protein;
complete cds
4 RC AA098874 ESTs
zn25b3.s1 Stratagene neuroepithelium NT2RAM1
2 937234 Homo Sapiens
C AA101056 cDNA clone IMAGE:548429 3' similar to contains
L1.b3 L1 repetitive
element ;, mRNA sequence
1 RC AA102746 ESTs; Moderately similar to cytotoxic ligand
TRAIL receptor [H.sapiens]
2 RC AA114250 Homo Sapiens mRNA for KIAA512 protein;
s complete cds
2 5 4 RC AA126561 ESTs
s
ESTs; Weakly similar to !!!! ALU SUBFAMILY
4 RC AA128980 J WARNING ENTRY 1111
i [H.sapiens)
ESTs; Highly similar to 6S RIBOSOMAL PROTEIN
4 RC AA129757 L22 [Rattus
norvegicus]
4 RC AA129921 ESTs
2 RC AA133331 Homo sapiens mRNA for KIAA741 protein;
complete cds
3 0 2 RC AA135958 ESTs
4 RC AA136524 ESTs
s
4 RC AA147044 ESTs; Weakly similar to transformation-related
protein [H.sapiens]
4 RC AA148885 ESTs
4 RC AA150043 ESTs
3 5 2 RC AA151621 ESTs
4 RC AA155743 ESTs
84
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Accession
Cluster#/ Gene Description
PROBESET
2 RC AA156335 ESTs
4 RC AA156336 Homo sapiens nuclear receptor co-repressor
N-CoR mRNA; complete cds
2 RC AA159181 ESTs
2 RC AA159825 ESTs
2 RC AA234185 ESTs
4 RC AA234929 ESTs
1 RC AA234935 ESTs
4 RC AA236359 ESTs
2 RC AA236466 ESTs
2 RC AA236535 ESTs
4 RC AA236935 Human normal keratinocyte mRNA
s
2 RC AA236942 ESTs
4 RC AA237018 ESTs
2 RC AA237025 ESTs
2 RC AA242751 Homo Sapiens mRNA for KIAA93 protein; partial
cds
3 RC AA242760 ESTs
3 RC AA242763 Homo sapiens Cdc14B1 phosphatase mRNA;
complete cds
ESTs; Weakly similar to !!!! ALU SUBFAMILY
2 RC AA242809 J WARNING ENTRY !!!!
[H.sapiens]
ESTs; Highly similar to SERINE/THREONINE-PROTEIN
3 RC AA243133 KINASE IPL1
[Saccharomyces cerevisiae]
2 0 4 RC AA243495 ESTs
3 RC AA243706 ESTs
zs6e2.s1 NCI CGAP_GCB1 Homo sapiens cDNA
4 RC AA250848 clone IMAGE:68441 3',
mRNA sequence
2 RC AA250868 ESTs
4 RC AA251152 ESTs
2 5 2 RC AA251544 ESTs
s
4 RC AA251792 ESTs
4 RC AA252063 Homo Sapiens mRNA for PCDH7 (BH-Pcdh)c;
complete cds
3 RC AA252144 ESTs
4 RC AA252524 ESTs
3 0 3 RC AA253461 ESTs
4 RC AA255522 ESTs
2 RC AA256468 ESTs
4 RC AA256528 ESTs
2 RC AA257976 ESTs
3 5 4 RC AA258296 Homo Sapiens mRNA for KIAA579 protein;
partial cds
3 RC AA258409 H.sapiens gene from PAC 313L4; similar
to Myelin PO
2 RC AA258421 Homo sa iens clone 683 unknown mRNA; com
lete se uence
CA 02381699 2002-02-07
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Accession
Cluster#/ Gene Description
PROBESET
Human NAD+-dependent succinate-semialdehyde
3 RC AA262077 dehydrogenase
(SSADH) mRNA; 3' end
4 RC AA278650 ESTs
2 RC AA278766 ESTs
4 RC AA279667 natural killer-tumor recognition sequence
s
3 RC AA280791 eukaryotic translation initiation factor
5
4 RC AA280819 ESTs
4 RC AA280828 ESTs
4 RC AA282195 ESTs; Weakly similar to ORF YNL292w [S.cerevisiae]
2 RC AA283127 Homo sapiens clone LM1955 H15e3 gene; partial
s cds
2 RC AA284694 Homo sapiens CG1 mRNA; complete cds
3 RC AA291137 ESTs
3 RC AA291708 ESTs; Moderately similar to hypothetical
protein [H.sapiens]
Homo sapiens BAC clone 255A7 from 8q21
3 RC AA293495 containing NBS1 gene;
complete sequence
ESTs; Weakly similar to NADH-UBIQUINONE
4 RC AA347193 OXIDOREDUCTASE
CHAIN 4 [Caenorhabditis elegansj
4 RC AA398474 ESTs
s
4 RC AA398512 ESTs
2 RC AA400277 ESTs; Weakly similar to putative p15 [H.sapiensj
4 RC AA400896 ESTs
3 RC AA404494 CTP synthase
2 0 2 RC AA410345 ESTs; Weakly similar to A33 antigen precursor
[H.sapiens]
4 RC AA416733 ESTs; Weakly similar to neuronal thread
protein AD7c-NTP [H.sapiens]
4 RC AA425154 ESTs
4 RC AA426573 ESTs
2 RC AA431418 N-acetylglucosaminidase; alpha- (Sanfilippo
disease IIIB)
2 5 4 RC AA436182 ESTs
2 RC AA437099 ESTs
4 RC AA446585 ESTs
3 RC AA446887 ESTs
ESTs; Highly similar to HYPOTHETICAL 8.7
2 RC AA447224 KD PROTEIN IN
ERG7-NMD2 INTERGENIC REGION [Saccharomyces
cerevisiae]
3 0 2 RC AA447709 ESTs; Moderately similar to putative transcription
factor CA15 [H.sapiens]
4 RC AA453624 deoxynucleotidyltransferase; terminal
4 RC AA455044 ESTs
4 RC AA456045 ESTs
4 RC AA460454 ESTs; Weakly similar to KIAA512 protein
s [H.sapiens]
3 5 4 RC AA476494 ESTs; Weakly similar to KIAA512 protein
[H.sapiens]
4 RC AA476738 ESTs; Highly similar to FLI-LRR associated
protein-1 [M.musculus]
4 RC AA481422 Homo sapiens mRNA for H-2K binding factor-2;
complete cds
3 RC AA482269 inte ral membrane rotein 1
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Accession
Cluster#I Gene Description
PROBESET
2 RC AA482595 ESTs; Highly similar to p36
4 RC AA485084 ESTs
s
4 RC_AA485431 ESTs
s
4 RC AA489057 H.sapiens mRNA for nuclear protein SA-2
4 RC AA489638 ESTs
2 RC AA491000 ESTs
3 RC AA491250 ESTs
4 RC_AA505133 ESTs
4 RC AA598447 Homo sapiens exportin t mRNA; complete
cds
3 RC AA599243 ESTs
3 RC AA599574 ESTs
i
4 RC AA600153 DEK gene
4 RC AA609309 ESTs
ESTs; Highly similar to HYPOTHETICAL GTP-BINDING
4 RC AA609710 PROTEIN IN
PM14-PAC2 INTERGENIC REGION [Saccharomyces
cerevisiae)
4 RC AA610068 H.sapiens mRNA for PIBF1 protein; complete
1 RC AA621399 ESTs
Human 26S proteasome-associated pad1 homolog
4 RC AA621752 (POH1 ) mRNA;
complete cds
2 RC C21523 ESTs
2 RC D12160 ESTs; Weakly similar to unknown [H.sapiens]
2 0 4 RC_D19708 ESTs
2 RC D25801 ESTs; Highly similar to KIAA445 protein
[H.sapiens]
2 RC D45652 ESTs; Weakly similar to unknown [H.sapiens]
4 RC D60208 ESTs
f
3 RC D80504 zinc finger protein 198
s
2 5 2 RC F03010 ESTs; Weakly similar to ZINC FINGER PROTEIN
HRX [Homo sapiens]
ESTs; Weakly similar to !!!! ALU CLASS
4 RC F04247 A WARNING ENTRY !!!!
[H.sapiens]
ESTs; Weakly similar to !!!! ALU SUBFAMILY
4 RC F10966 J WARNING ENTRY !!!!
[H.sapiens]
Homo sapiens ribonuclease P protein subunit
4 RC F13700 p4 (RPP4) gene; complete
cds
4 RC H05063 ESTs
ESTs; Highly similar to ERYTHROPOIETIN
3 0 4 RC H16758 RECEPTOR PRECURSOR
[Homo Sapiens]
4 RC H17315 karyopherin alpha 1 (importin alpha 5)
s
4 RC H22556 PROTEIN TRANSLATION FACTOR SUIT HOMOLOG
ESTs; Highly similar to protein tyrosine
4 RC H22566 phosphatase epsilon cytoplasmic
isoform [H.sapiens]
4 RC H48459 Human mRNA for KIAA186 gene; complete cds
s
3 5 4 RC H53073 ESTs
2 RC H56559 Homo sapiens mRNA for KIAA61 protein; partial
s cds
3 RC H57957 ESTs
s
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Accession
Cluster#/ Gene Description
PROBESET
2 RC H64938 ESTs
s
2 RC H64973 ESTs
4 RC H69535 ESTs
ESTs; Moderately similar to alternatively
2 RC H73110 spliced product using exon 13A
[H.sapiens]
2 RC H81783 ESTs
1 RC H86259 Homo sapiens chromosome 19; cosmid 832611
2 RC H88353 ESTs; Weakly similar to reverse transcriptase
related protein [H.sapiens]
2 RC H88639 ESTs
4 RC H88675 ESTs
4 RC H93708 CLEAVAGE SIGNAL-1 PROTEIN
s
ESTs; Weakly similar to !!!! ALU SUBFAMILY
4 RC N22107 J WARNING ENTRY !!!!
[H.sapiens]
3 RC N24046 ESTs; Weakly similar to 6S RIBOSOMAL PROTEIN
L1 [Homo sapiens]
2 RC N27028 ESTs
2 RC N30205 ESTs; Weakly similar to hypothetical protein
[H.sapiens]
1 RC N30621 ESTs
4 RC N33258 Homo sapiens nuclear receptor co-repressor
N-CoR mRNA; complete cds
2 RC N33390 EST
2 RC N40180 EST; Weakly similar to putative p15 [H.sapiens]
2 RC N45198 EST
2 0 3 RC N45979 SH3 domain protein 1 B
s
2 RC N48325 EST
2 RC N48913 ESTs
4 RC N49394 Homo sapiens mRNA for KIAA716 protein;
complete cds
1 RC N50656 ESTs; Highly similar to mosaic protein
LR11 [H.sapiens]
4 RC N50721 kinesin family protein 3B
4 RC N53143 ESTs
2 RC N53359 ESTs
4 RC N55326 ESTs
yv5c2.s1 Soares fetal liver spleen 1 NFLS
2 RC N55493 Homo sapiens cDNA clone
IMAGE:246146 3', mRNA sequence
3 0 4 RC N57493 EST
2 RC N62955 ESTs; Weakly similar to ankyrin G [H.sapiens]
4 RC N63520 EST; Weakly similar to mariner transposase
[H.sapiens]
4 RC N63604 ESTs
2 RC N64166 frizzled (Drosophila) homolog 7
3 5 2 RC N64168 ESTs
2 RC N64191 ESTs
ESTs; Weakly similar to !!!! ALU CLASS
4 RC N66845 B WARNING ENTRY !!!!
[ H.sapiens]
4 RC N67135 ESTs
88
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Accession
Cluster#/ Gene Description
PROBESET
2 RC N67295 ESTs
4 RC N68399 H2B histone family; member N
4 RC N68963 ESTs
4 RC N69331 peptidylprolyl isomerase C (cyclophilin
C)
2 RC N70777 ESTs
1 RC N71364 ESTs; Weakly similar to transformation-related
s protein [H.sapiens]
4 RC N71545 ESTs; Moderately similar to hypothetical
s protein [H.sapiens]
2 RC N71571 ESTs
4 RC N74456 EST
4 RC N75594 ESTs
2 RC N79035 EST
2 RC N80279 ESTs; Highly similar to (def)ine not available
4239677) [H.sapiens]
4 RC N91797 ESTs
4 RC N92454 karyopherin (importin) beta 1
4 RC N94581 actin; beta
4 RC N94746 ESTs
4 RC N98238 ESTs
4 RC 802384 EST
ESTs; Weakly similar to !!!! ALU CLASS
2 RC 816833 F WARNING ENTRY !!II
[H.sapiens]
2 0 3 RC 841828 myosin VA (heavy polypeptide 12; myoxin)
s
2 RC 843203 ESTs
4 RC 846395 ESTs; Moderately similar to Unknown gene
product [H.sapiens]
2 RC 858863 ESTs
2 RC 878248 ESTs
2 5 4 RC T11483 ESTs
4 RC T16896 ESTs
2 RC T23820 cyclin T2
4 RC T30222 ESTs; Weakly similar to tetracycline transporter-like
protein [M.musculus]
4 RC W15275 ESTs
s
3 0 2 RC W38194 Accession not listed in Genbank
3 RC W42414 ESTs
s
4 RC W46577 H.sapiens mRNA for ESM-1 protein
s
4 RC W49632 Human clone 2398 mRNA sequence
s
2 RC W57613 ESTs
3 5 2 RC W57759 EST
4 RC W61118 ESTs
4 RC W65344 ESTs; Highly similar to ICH-2 PROTEASE
PRECURSOR [Homo Sapiens]
2 RC W69216 ESTs
ESTs; Weakly similar to mitochondria) inner
2 RC W69379 membrane protease 1
[ S.cerevisiae]
4 0 4 RC W86728 ESTs
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Accession
Cluster#/ Gene Description
PROBESET
4 RC 238499 ESTs; Weakly similar to protein phosphatase
[H.sapiens]
2 RC 238630 Homo sapiens 1kD protein (BC1 ) mRNA;
complete cds
4 RC 239494 ESTs
4 RC 239623 ESTs
3 RC 240071 BMX non-receptor tyrosine kinase
s
2 RC 240174 EST
2 RC 240182 EST
2 RC 240904 EST
4 AFFX-BioB-3
4 AFFX-BioC-3
3 AFFX-DapX-5
1 AFFX-LysX-M
3 RC AA166965 ESTs
3 RC AA167500 EST
1 RC AA169599 ESTs
s
3 RC AA171724 ESTs
2 RC AA171739 ESTs
3 RC AA177105 ESTs
2 RC AA182626 ESTs
ESTs; Highly similar to cell cycle progression
2 0 3 RC AA186324 restoration 8 protein
[H.sapiens]
1 RC AA192099 zinc finger protein 148 (pHZ-52)
ESTs; Moderately similar to !!!! ALU SUBFAMILY
3 RC AA192173 SC WARNING ENTRY
!!!! (H.sapiens]
3 RC AA192415 EST
3 RC AA192553 ESTs; Moderately similar to RGC-32 [R.norvegicus]
2 5 3 RC AA194851 ESTs
3 RC AA195520 ESTs
s
ESTs; Moderately similar to !!!! ALU SUBFAMILY
3 RC AA196300 SO WARNING ENTRY
!!!! [H.sapiens]
3 RC AA196517 Lon protease-like protein
3 RC AA196549 ESTs
zq9a3.s1 Stratagene muscle 93729 Homo
3 sapiens cDNA clone
0 C AA196721 IMAGE:629164 3' similar to TR:G746415
6746415 I KAPPA BR. ;, mRNA
sequence
ESTs; Weakly similar to !!!! ALU SUBFAMILY
3 RC AA196729 J WARNING ENTRY !!!!
i [H.sapiens]
ESTs; Moderately similar to RETROVIRUS-RELATED
1 RC AA196979 PROTEASE
(H.sapiens]
2 RC AA206828 ESTs; Weakly similar to ubiquitous TPR
motif; Y isoform [H.sapiens]
3 RC AA207123 immunoglobulin supertamily; member 3
3 5 1 RC AA214539 ESTs
i
3 RC AA226914 TR2 nuclear hormone rece for
s
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Accession
Cluster#/ Gene Description
PROBESET
3 RC AA227260 Zic family member 3 (odd-paired Drosophila
homolog; heterotaxy 1 )
3 RC AA227469 EST
ESTs; Highly similar to CALCIUM/CALMODULIN-DEPENDENT
3 RC AA233122 PROTEIN KINASE TYPE II DELTA CHAIN (Rattus
norvegicus]
3 RC AA233334 Homo sapiens josephin MJD1 mRNA; cds
s
Homo sapiens zinc finger protein 216 splice
3 RC AA233347 variant 2 (ZNF216) mRNA;
complete cds
1 RC AA233519 ESTs; Weakly similar to neuronal thread
protein AD7c-NTP [H.sapiens]
1 RC AA233714 Apg12 (autophagy; yeast) homolog
1 RC AA233796 ESTs
1 RC AA235050 ESTs
f
ESTs; Weakly similar to Wiscott-Aldrich
1 RC AA235704 Syndrome protein homolog
[M.musculus]
3 RC AA236031 ESTs
1 RC AA236352 ESTs
1 RC AA236390 ESTs
s
1 RC AA236453 ESTs
3 RC AA243370 EST
2 RC AA250947 ESTs
3 RC AA251083 ESTs
3 RC AA251113 ESTs
4 RC AA251973 ESTs
2 0 3 RC AA252023 ESTs; Moderately similar to (defline not
available 397874) [H.sapiens]
1 RC AA252414 ESTs
1 RC AA252650 protein kinase; mitogen-activated; kinase
7 (MAP kinase kinase 7)
3 RC AA255523 ESTs
3 RC AA258128 ESTs
2 5 3 RC AA262105 Human mRNA for KIAA331 gene; complete cds
1 RC AA262107 ESTs
1 RC AA262235 ESTs
zs8b3.s1 NCI CLAP GCB1 Homo sapiens cDNA
3 RC AA278298 clone IMAGE:73757 3',
mRNA sequence
1 RC AA278529 ESTs; Highly similar to serine/threonine
i protein kinase [H.sapiens]
3 0 1 RC AA278721 ESTs
1 RC AA280036 ESTs
1 RC AA280648 ESTs; Weakly similar to rab-related GTP-binding
protein [H.sapiens]
1 RC AA280738 ESTs
3 RC AA280794 ESTs
3 5 1 RC AA280837 ESTs
ESTs; Moderately similar to alternatively
1 RC AA280886 spliced product using exon 13A
[H.sapiens]
1 RC AA280934 ESTs
1 RC AA281535 Homo sa iens mRNA for KIAA879 rotein; com
lete cds
91
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Accession
Cluster#/ Gene Description
PROBESET
Homo Sapiens basic transcription factor
4 2 p44 (btf2p44) gene; partial cds;
C AA281797 neuronal apoptosis inhibitory protein (naip)
s and survival motor neuron
protein (smn) genes; complete cds
1 RC AA282047 ESTs
1 RC AA283002 Human zinc finger protein (SRE-ZBP) mRNA;
3' end
3 RC AA283709 ESTs
ESTs; Weakly similar to X-linked retinopathy
1 RC AA283902 protein {C-terminal; clone
XEH.Bc} [H.sapiens]
Human DNA from chromosome 19-specific cosmid
1 RC AA284108 F25965; genomic
sequence
Human DNA sequence from clone 71 L16 on
1 chromosome Xp11. Contains
C AA284109 a probable Zinc Finger protein (pseudo)gene;
an unknown putative gene;
a pseudogene with high similarity to part
of antigen KI-67; a pu
1 RC AA284371 Homo sapiens clone 23967 unknown mRNA;
partial cds
3 RC AA284744 ESTs
f
1 RC AA284784 ESTs
1 RC AA284840 ESTs
1 RC AA286844 ESTs
3 RC AA287032 ESTs
1 RC AA287038 EST
1 RC AA287546 ESTs
1 RC AA287553 ESTs
s
ESTs; Weakly similar to !!!! ALU CLASS
3 RC AA287556 B WARNING ENTRY !Ill
(H.sapiens]
1 RC AA287564 ribosomal protein L37
1 RC AA291015 CDC7 (cell division cycle 7; S. cerevisiae;
s homology-like 1
2 0 3 RC AA291716 EST
1 RC AA291749 ESTs
s
1 RC AA293656 EST
1 RC AA302430 ESTs
3 RC AA302809 EST
2 5 1 RC AA302820 purinergic receptor P2X; ligand-gated ion
s channel; 4
1 RC AA310499 ESTs
1 RC AA321890 EST24442 Cerebellum II Homo sapiens cDNA
3' end, mRNA sequence
1 RC AA340589 EST
1 RC AA340622 ESTs
3 0 1 RC AA342457 ESTs
i
3 RC AA342828 glycoprotein V (platelet)
s
1 RC AA342864 ESTs
1 RC AA342973 ESTs
1 RC AA346495 ESTs
3 5 1 RC AA347573 Homo sapiens KIAA45 mRNA; complete cds
1 RC AA347614 ESTs
1 RC AA347717 ESTs
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Accession
Cluster#/ Gene Description
PROBESET
1 RC AA348913 ESTs
1 RC AA349647 EST
1 RC AA349773 ESTs
ESTs; Moderately similar to alternatively
1 RC AA350541 spliced product using exon 13A
s [H.sapiens]
1 RC AA357159 EST
i
3 RC AA357172 ESTs
i
1 RC AA369856 Human hVps4lp (hVPS41) mRNA; alternative
s splice variant; partial cds
1 RC AA370132 EST
1 RC AA370472 ESTs
s
1 RC AA370867 ESTs
3 RC AA377296 ESTs
4 RC AA383902 ESTs
3 RC AA385934 EST; Highly similar to predicted using
Genefinder [C.elegans]
3 RC AA386255 EST
1 5 1 RC AA386260 EST
ESTs; Highly similar to MEMBRANE GLYCOPROTEIN
2 RC AA386266 M6-B [Mus
musculus]
1 RC AA398014 ESTs
3 RC AA398222 ESTs
1 RC AA398235 ESTs
2 0 3 RC AA398348 ESTs
3 RC AA398482 ESTs
1 RC AA398504 ESTs
1 RC AA398505 ESTs
1 RC AA398507 ESTs
ESTs; Weakly similar to !!!! ALU SUBFAMILY
2 5 1 RC AA398523 SO WARNING ENTRY !Ill
[H.sapiens]
1 RC AA398625 ESTs
4 RC AA398632 ESTs
3 RC AA398633 ESTs
3 RC AA398894 ESTs
3 0 3 RC AA398895 EST
1 RC AA398900 ESTs
1 RC AA398904 EST
1 RC AA399122 ESTs; Weakly similar to mitochondria)
citrate transport protein [H.sapiens]
3 RC AA399371 ESTs
3 5 1 RC AA399373 ESTs; Highly similar to KIAA568 protein
[H.sapiens]
1 RC AA399441 ESTs
3 RC AA399636 ESTs
1 RC AA399640 ESTs
1 RC AA399680 ESTs
93
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Accession
Cluster#/ Gene Description
PROBESET
3 RC AA400080 EST
1 RC AA400262 ESTs
1 RC AA400725 ESTs
3 RC AA400748 ESTs
1 RC AA400780 ESTs
zv65b9.s1 Soares_total_fetus Nb2HF8 9w
3 RC AA40163 Homo sapiens cDNA clone
1 IMAGE:758489 3', mRNA sequence
1 RC AA401688 ESTs
1 RC AA401695 EST
3 RC AA402227 ESTs; Moderately similar to N-tropomodulin
[R.norvegicus]
3 RC AA402329 ESTs
1 RC AA402398 ESTs
1 RC AA402449 EST
1 RC AA402468 ESTs
2 RC AA403268 ESTs
s
2 RC AA403314 ESTs
1 RC AA404229 EST
1 RC AA404260 ESTs
1 RC AA404271 Human glutamate/kainate receptor subunit
(EEA3) mRNA; complete cds
3 RC AA405026 ESTs
2 0 1 RC AA405182 ESTs
ESTs; Moderately similar to alternatively
1 RC AA405237 spliced product using exon 13A
[H.sapiens]
3 RC AA406061 EST
1 RC AA406063 ESTs
1 RC AA406070 EST
2 5 1 RC AA406137 EST
1 RC AA406335 ESTs
1 RC AA411801 Human mRNA for KIAA37 gene; complete cds
1 RC AA411804 ESTs
1 RC AA411833 ESTs; Highly similar to (defline not available
4521278) [H.sapiens]
3 0 1 RC AA412219 ESTs
3 RC AA412259 ESTs
2 RC AA412497 Human Line-1 repeat mRNA with 2 open reading
frames
1 RC AA412498 ESTs
1 RC AA416586 ESTs
3 5 1 RC AA416867 EST
1 RC AA416874 ESTs
1 RC AA421133 ESTs
1 RC AA421138 EST
ESTs; Highly similar to ELONGATION FACTOR
4 RC AA422079 G; MITOCHONDRIAL
PRECURSOR [Rattus norvegicusJ
4 0 1 RC AA423837 ESTs
1 RC AA424328 ESTs
1 RC AA424339 ESTs
94
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Accession
Cluster#/ Gene Description
PROBESET
3 RC AA424469 ESTs
s
1 RC AA424502 ESTs
3 RC AA425004 ESTs
1 RC AA425734 ESTs; Weakly similar to neuronal thread
protein AD7c-NTP [H.sapiens]
1 RC AA425887 ESTs
3 RC AA426456 ESTs
3 RC AA427396 ESTs
1 RC AA427555 Human mRNA for KIAA23 gene; complete cds
3 RC AA428218 ESTs
1 0 3 RC AA428242 ESTs
1 RC AA428281 EST
3 RC AA428865 EST
3 RC AA428994 ESTs
1 RC AA429666 ESTs
3 RC AA430181 ESTs
1 RC AA430184 Human putative ATP/GTP-binding protein
s (HEAB) mRNA; complete cds
3 RC AA431288 CD3D antigen; delta polypeptide (TiT3 complex)
s
1 RC AA431293 ESTs
3 RC AA431478 ESTs
2 0 3 RC AA431492 EST
1 RC AA431732 EST
3 RC AA432278 EST
4 RC AA434411 ESTs
3 RC AA435512 ESTs
i
2 5 1 RC AA435698 ESTs
1 RC AA435711 Homo sapiens mRNA for KIAA712 protein;
complete cds
3 RC AA435815 Clk-associating RS-cyclophilin
s
3 RC AA435842 ESTs
3 RC AA436475 ESTs
3 0 3 RC AA436489 ESTs
3 RC AA442060 ESTs
1 RC AA442079 EST
3 RC AA443151 ESTs
4 RC AA446133 ESTs
3 5 1 RC AA447145 Homo sapiens KIAA399 mRNA; partial cds
3 RC AA447398 EST
1 RC AA447643 ESTs
1 RC AA447742 dynein; axonemal; heavy polypeptide 17-like
s
zw96c1.s1 Soares_total_fetus Nb2HF8 9w
3 RC AA44822 Homo sapiens cDNA clone
6 IMAGE:784818 3', mRNA sequence
4 0 1 RC AA448825 EST
1 RC AA449444 ESTs
3 RC AA450087 re ulator of Gz-selective rotein si nalin
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Accession
Cluster#/ Gene Description
PROBESET
3 RC AA450211 EST
1 RC AA450244 ESTs
3 RC AA452123 ESTs; Weakly similar to Tcp-1 [M.musculus]
3 RC AA452155 zinc finger protein 198
3 RC AA452156 EST
3 RC AA453036 ESTs
3 RC AA453526 ESTs
3 RC AA454085 EST
3 RC AA454103 ESTs
1 RC AA454642 ESTs
1 RC AA454935 ESTs
3 RC AA456323 ESTs
3 RC AA457395 EST
aa26c7.s1 NCI_CGAP_GCB1 Homo sapiens cDNA
3 RC AA458850 clone IMAGE:81438
3' similar to contains L1.t3 L1 repetitive
element ;, mRNA sequence
3 RC AA459662 EST
Homo Sapiens 3-hydroxyisobutyryl-coenzyme
3 RC AA459668 A hydrolase mRNA;
complete cds
ESTs; Weakly similar to The KIAA191 gene
1 RC AA459679 is expressed ubiquitously.
s [H.sapiens]
1 RC AA459702 ESTs
4 RC AA460017 ESTs
f
2 0 3 RC AA460324 ESTs
3 RC AA461509 ESTs; Weakly similar to hypothetical protein
II [H.sapiens]
3 RC AA464414 ESTs
i
1 RC AA464428 ESTs
3 RC AA470084 ESTs
2 5 3 RC AA476606 ESTs
s
3 RC AA478521 ESTs
ESTs; Moderately similar to !!!! ALU SUBFAMILY
3 RC AA478523 J WARNING ENTRY !!!!
[H.sapiens]
3 RC AA479949 ESTs
3 RC AA481252 RAS-LIKE PROTEIN TC21
3 0 1 RC AA485351 ESTs; Weakly similar to predicted using
Genefinder [C.elegans]
1 RC AA487264 ESTs
1 RC AA489072 Homo Sapiens mRNA for KIAA87 protein;
complete cds
1 RC AA489630 Homo sapiens mRNA for KIAA665 protein;
complete cds
2 RC AA490225 ESTs
3 5 3 RC AA490227 ESTs
3 RC AA490255 ESTs
1 RC AA490890 ESTs
2 RC AA490916 ESTs
s
3 RC AA490925 Homo sa iens laforin EPM2A mRNA; artial
cds
96
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Accession
Cluster#/ Gene Description
PROBESET
1 RC AA490955 ESTs; Weakly similar to bullous pemphigoid
antigen [M.musculus]
1 RC AA495812 ESTs
3 RC AA495824 ESTs
1 RC AA496369 ESTs
3 RC AA504125 ESTs
s
1 RC AA521473 Human brain secretory protein hSec1 p (HSEC1
) mRNA; complete cds
1 RC AA598440 ESTs
3 RC AA598899 ESTs
i
3 RC AA599244 Homo sapiens mRNA for KIAA53 protein; partial
cds
1 RC AA599694 Human mRNA for KIAA133 gene; complete cds
s
1 RC AA600037 ESTs
3 RC AA609135 EST
1 RC AA609582 Homo sapiens p6 katanin mRNA; complete
cds
3 RC AA609684 ESTs
3 RC AA609839 ESTs
1 RC AA609862 Homo sapiens mRNA for RBP-MS/type 3; complete
cds
4 RC AA620423 EST
3 RC AA620747 EST
1 RC AA621364 ESTs
2 0 2 RC C20653 ESTs '
3 RC D20085 ESTs
1 RC D20749 ESTs
2 RC D51285 ESTs
s
4 RC D59972 ESTs
i
2 5 4 RC F04112 ESTs
f
2 RC F13604 ESTs
1 RC H01662 ESTs
1 RC H05135 ESTs
i
3 RC H12245 splicing factor; arginine/serine-rich 7
(35kD)
3 0 1 RC H22842 EST
1 RC H30894 ESTs
2 RC H43442 Human mRNA for KIAA28 gene; partial cds
s
3 RC H45996 ESTs
2 RC H69281 ESTs
i
3 5 3 RC H69485 ESTs
f
1 RC H69899 ESTs; Moderately similar to unknown [H.sapiens]
4 RC H70627 ESTs
s
1 RC H73050 Rhesus blood group; D antigen
s
1 RC H73260 ESTs
4 0 1 RC H77531 HIR (histone cell cycle regulation defective;
s S. cerevisiae) homolog A
2 RC H80552 EST
4 RC H80737 lysyl oxidase
s
1 RC H93412 ESTs
3 RC H94892 v-ral simian leukemia viral onto ene homolo
s A ras related
97
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Accession
Cluster#/ Gene Description
PROBESET
4 RC H95643 neurotrophic tyrosine kinase; receptor;
s type 1
2 RC H96552 ESTs
ESTs; Highly similar to G protein-coupled
4 RC H97146 receptor kinase 6; splice variant
B [H.sapiens]
2 RC H99131 ESTs
s
1 RC H99462 ribosomal protein; mitochondrial; L12
s
1 RC H99837 ESTs
s
ESTs; Highly similar to TUBULIN GAMMA CHAIN
2 RC N22140 [Euplotes
octocarinatus]
2 RC N22197 ESTs
1 RC N23756 Human mRNA for KIAA238 gene; partial cds
s
2 RC N24134 eukaryotic translation initiation factor
1A; Y chromosome
4 RC N24195 Homo sapiens mRNA for RanBPM; complete
cds
1 RC N26739 CAAX box 1
2 RC N27098 EST
1 RC N27637 ESTs
4 RC N33090 ESTs; Weakly similar to translation initiation
factor [H.sapiens]
1 RC N35967 ESTs
Homo sapiens chaperonin containing t-complex
1 RC N38959 polypeptide 1; beta
f subunit (Cctb) mRNA; complete cds
2 RC N39069 ESTs
1 RC N46441 ESTs
2 0 2 RC N48270 ESTs
f
2 RC N48365 ESTs
s
2 RC N51316 ESTs
1 RC N51499 ESTs
s
4 RC N53976 ESTs
2 5 2 RC N54157 ESTs
2 RC N54300 ESTs
1 RC N54831 ESTs; Weakly similar to neuronal thread
protein AD7c-NTP [H.sapiens]
2 RC N59849 ESTs
4 RC N62132 ESTs
3 0 1 RC N62375 EST
4 RC N63138 ESTs
1 RC N63172 cell division cycle 42 (GTP-binding protein;
25kD)
Homo sapiens DNA sequence from PAC 434014
on chromosome
2 1q32.3.-41. Contains the HSD11B1 gene for
C N63772 Hydroxysteroid (11-beta)
Dehydrogenase 1; the ADORA2BP adenosine
A2b receptor LIKE
pseudogene; the IRF
2 RC N63787 ESTs
za11c1.s1 Soares fetal liver spleen 1NFLS
3 5 2 RC N68168 Homo sapiens cDNA clone
IMAGE:292224 3', mRNA sequence
2 RC N68201 ESTs; Weakly similar to hypothetical protein
[H.sapiens]
2 RC N68300 ESTs
98
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Accession
Cluster#/ Gene Description
PROBESET
1 RC N68321 solute carrier family 2 (facilitated glucose
transporter); member 3
2 RC N69575 EST
2 RC N75007 ESTs
1 RC N75542 ESTs
2 RC N90066 Homo sapiens clone 24689 mRNA sequence
1 RC N91246 ESTs
ESTs; Weakly similar to cyclic nucleotide-gated
1 RC N92751 channel beta subunit
[R.norvegicus]
2 RC N93214 ESTs
s
2 RC N99148 ESTs; Highly similar to MKR2 PROTEIN [Mus
musculus]
ESTs; Weakly similar to HYPOTHETICAL PROTEIN
4 RC 807876 H11723
[Haemophilus influenzae]
1 RC 810865 alpha-fetoprotein
f
2 RC 811056 ESTs
2 RC 811488 ESTs
1 RC 822947 ESTs
2 RC 823930 ESTs
s
1 RC 826589 ESTs
f
4 RC 837588 GDS-related protein
s
2 RC 837613 ESTs
1 RC 838398 Homo sapiens clone 23758 mRNA sequence
2 0 2 RC 839179 ESTs
f
1 RC 840923 ESTs
1 RC 841179 Human mRNA for KIAA328 gene; partial cds
2 RC 841294 ESTs
s
1 RC 842307 early development regulator 2 (homolog
f of polyhomeotic 2)
2 5 1 RC 843189 EST
f
3 RC 843306 ESTs
1 RC 844357 ESTs
1 RC 844519 EST; Moderately similar to Pro-Pol-dUTPase
polyprotein [M.musculus]
yg38g4.s1 Soares infant brain 1 NIB Homo
2 RC 845088 Sapiens cDNA clone
IMAGE:34896 3', mRNA sequence
3 0 2 RC 847948 ESTs
i
1 RC 851524 ESTs
1 RC 854950 ESTs
1 RC 855241 EST
1 RC 859585 ESTs
3 5 1 RC 860044 ESTs
2 RC 860872 ESTs
1 RC 866690 ESTs
2 RC 867266 exostoses (multiple)-like 1
s
1 RC 873588 ESTs
4 0 3 RC 879403 ESTs
99
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Accession
Cluster#/ Gene Description
PROBESET
1 RC 887647 ESTs
2 RC 893622 ESTs
4 RC 899599 heterogeneous nuclear ribonucleoprotein
s U (scaffold attachment factor A)
4 RC 899612 ESTs
FB14D6 Fetal brain, Stratagene Homo sapiens
1 RC T02888 cDNA clone FB14D6
3'end, mRNA sequence
1 RC T03170 EST
hbc313 Human pancreatic islet Homo Sapiens
2 RC T10465 cDNA clone hbc313 3'end,
mRNA sequence
1 RC T15418 ESTs
f
1 RC T15597 Homo sapiens mRNA for KIAA661 protein;
f complete cds
2 RC T15652 ESTs
i
2 RC T16898 ash2 (absent; small; or homeotic; Drosophila;
s homology-like
1 RC T26644 ESTs; Weakly similar to zinc finger protein
i ZNF139 (H.sapiens]
2 RC T40841 ESTs
yb15c11.s1 Stratagene placenta (#937225)
1 Homo sapiens cDNA clone
C T47566 i IMAGE:71252 3' similar to similar to gb:Z2157
ELONGATION FACTOR
1-DELTA (HUMAN), mRNA sequence
ESTs; Moderately similar to EA22 GENE PROTEIN
2 RC T50116 [Bacteriophage
lambda]
2 RC T50145 FSHD region gene 1
s
ESTs; Moderately similar to !!!! ALU SUBFAMILY
2 RC T58615 J WARNING ENTRY !!!!
[H.sapiens]
1 RC T59940 ESTs
f
4 RC T63595 ESTs
ydlc2.s1 Soares fetal liver spleen 1NFLS
2 0 2 RC T64891 Homo sapiens cDNA clone
IMAGE:66722 3', mRNA sequence
2 RC T64924 ESTs
2 RC T64933 ESTs; Weakly similar to hypothetical protein
r [H.sapiens]
yc3f5.s1 Stratagene liver (#937224) Homo
2 RC T68875 sapiens cDNA clone
IMAGE:8229 3', mRNA sequence
2 RC T69027 ESTs
yc19d3.s1 Stratagene lung (#93721 ) Homo
3 RC T69924 Sapiens cDNA clone
I MAGE:81125 3', mRNA sequence
3 RC T70353 ESTs
ESTs; Weakly similar to PUTATIVE MITOCHONDRIAL
1 RC T79780 CARRIER
s YBR291 C [Saccharomyces cerevisiae]
2 RC T79951 ESTs
3 RC T80174 ESTs
s
3 0 3 RC T80622 ESTs; Weakly similar to envelope protein
RIC-7 [H. sapiens]
1 RC T85352 ESTs
1 RC T85373 ESTs
100
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Accession
Cluster#/ Gene Description
PROBESET
2 RC T86284 ESTs; Weakly similar to transformation-related
protein [H.sapiens]
Homo sapiens E2F-related transcription
1 RC T89579 factor (DP-1 ) mRNA; complete
s cds
3 RC T90360 ESTs
2 RC T94328 ESTs
i
ye4a3.s1 Soares fetal liver spleen 1 NFLS
1 Homo Sapiens cDNA clone
C T95590 IMAGE:12172 3' similar to gb~M1817~IGURRAA
Iguana iguana 5S
(rRNA);, mRNA sequence
4 RC T97257 ESTs; Weakly similar to hypothetical protein
f [H.sapiens]
2 RC T97599 ESTs
i
2 RC T97620 ESTs; Weakly similar to unknown [H.sapiens]
4 RC T97775 ESTs
3 RC T98152 fibrillin 2
1 RC W31479 ESTs
1 RC W37999 ESTs
2 RC W38240 Accession not listed in Genbank
2 RC W40150 human chromosome-associated polypeptide
(bamacan)
2 RC W45435 Homo sapiens mRNA for KIAA784 protein;
partial cds
2 RC W58202 ESTs
1 RC W58344 ESTs
2 RC W58650 ESTs
Human DNA sequence from clone 1189824 on
chromosome Xq25-26.3.
4 Contains NADH-Ubiquinone Oxidoreductase
C W68736 MLRQ subunit (EC 1.6.5.3;
EC 1.6.99.3; CI-MLRQ); Tubulin Beta and
Proto-oncogene
Tyrosine-protein
2 0 2 RC W69106 ESTs
2 RC W69111 ESTs
1 RC W69385 H.sapiens NuMA gene (Clone T33)
s
3 RC W69399 ATPase; Ca++ transporting; plasma membrane
s 1
3 RC W69459 ESTs
2 RC W72424 S1 calcium-binding protein A9 (calgranulin
B)
2 RC W72724 ESTs
2 RC W72834 ESTs
1 RC W73955 Homo Sapiens chromosome 19; cosmid 826445
2 RC W74701 ESTs
3 0 2 RC W76540 ESTs
2 RC W79397 ESTs
ESTs; Weakly similar to synapse associated
2 RC W85888 protein sap47-2
[D.melanogaster]
2 RC W86038 ESTs
2 RC W86881 ESTs
3 5 2 RC W87804 ESTs
zh7b5.s1 Soares fetal_liver_spleen_1NFLS
2 RC W88942 S1 Homo sapiens cDNA
clone IMAGE:417393 3', mRNA se uence
101
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Accession
Cluster#/ Gene Description
PROBESET
3 RC_W90022 ESTs; Highly similar to LECT2 precursor
[H.sapiens]
2 RC W92272 Homo sapiens zinc-finger helicase (hZFH)
mRNA; complete cds
TUMOR NECROSIS FACTOR-INDUCIBLE PROTEIN
2 RC W92764 TSG-6
s PRECURSOR
2 RC_W93040 ESTs
3 RC W93092 neutral sphingomyelinase (N-SMase) activation
associated factor
2 RC W93227 EST
2 RC W93523 ESTs
2 RC W93659 ESTs
2 RC W94003 ESTs
s
2 RC W94401 ESTs
s
2 RC W94688 Homo sapiens mRNA for perilipin; complete
cds
2 RC W94787 ESTs
s
2 RC 238294 ESTs
s
3 RC 238311 ESTs
2 RC 238465 ESTs
s
2 RC 238525 ESTs
s
2 RC 238538 ESTs
f
2 RC 238551 ESTs
s
cat+-dependent activator protein for secretion;
2 RC 238783 Ca2+-regulated
s cytoskeletal protein (CAPS)
2 0 2 RC 239113 ESTs
4 RC 239255 ESTs
f
2 RC 239591 EST
2 RC 239783 ESTs
s
ESTs; Highly similar to NADH-CYTOCHROME
2 RC 239920 B5 REDUCTASE [Bos
taurus]
2 5 2 RC 240166 ESTs
f
3 RC 240388 ESTs
s
2 RC 240646 ESTs
2 RC 241697 ESTs
2 RC 299349 ESTs
3 0 2 RC 299394 ESTs~ Weakl similar to transformation-related
s rotein H.sa iens
102
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Table 2
0 o
X ~j
U v U
U x o
U v U ~
~
Z Z
(~ ~ fB (p ~
f0 -
d d ~ a
d
d .. a Q
Q ~
E- H- E- f- H f- H H H
Z Z Z Z Z Z Z >- Z y- ~ Z Z Z Z ~- Z ~ Z Z ~ Z ~ Z
~- >- Z Z
> a~ x
U v U
a
- - - - - _ ~ _ .n
a~ a~ a~ a~ a~ m a~ a~ a~
a ~
> >, >. >, > >, >, T
~. f- H H , >. H f- H
f- f- H
Z 7- Z Z Z Z Z Z ~ Z ~ ~ >- Z Z Z Z ~ Z Z Z ~ Z Z ~ Z >- Z
Z ~ Z Z r 7- Z Z Z Z ~ ~ ~ Z Z Z ~ Z Z Z Z >- J- Z Z ~ Z Z
f6
t
C L f' f~0 N
O Y U
d '00 _Q. w w a0 O
N O N 41 " ~ C
d ~ C C ~- O
O) .C d ~ N
O O N >' O "_O _a y L N f0
o a c o N
p ~ .U M V N N ~ ~ ~ ~ v E
O
L X O O 7 ~ ~ d N C ~ ~ N
N a7 v
Y Y " Q7 L _ ~ N l' O O ~ ~ a f9
d (O ca M ~ y C N C7 a d C ~ N O O
L L ~ ~ ' ~ O N N ~ ~ ~ ~ 'D N =' ~ U U
C C 'p O J J fD (O N N 07
w ~ O v NN~CC 10 c4 C~ _R .L-T.
7 d C :D ~ _ ' .. v- ~V ~V
CCCC _CNNCN~NO O.a~O~O~ICOO aOnL
~7 O f' m L y U U C ~ ~ L N N N Q ~ a Q O O
ao _a~ o_ a o _a y .. o ~ o - T _T v o o m ~° ~° ~ U U :__ m a~
m ..
N ~ N ~U ~U ~ L + d U O O O L L . _ C C
c = N N C ~ Q c Y c~ a a a a c_ ~ ~ ~ a c X
N O C C ~ m ~7 U N N ~ ~ U F- O O U N N N L L C 01 U ~ Q N In
c c ~' ... ~ m ~ y Z c ~ ~ c o~ a~ o io m m ~ ~ m o~ ~c a~ ~ w a~ a~ ~'
u' 0 0 o m ~ ~ E ~ c .~ c c '~ m m a v v' ~ ~ ~ ~ '~ m c ~ _> _>
Q U 'N ~N c0 ca C N O ~~ N Y ~~ p O X C C C L L C C C d O ~ O p~ N U U U
VJ C O U ~ ~ ~ L >, Q d ~ ~ ~U E C d 'O_ ~d ~ .~1 ~N N d L M t0 ~ ~ ~ N C C C
(D U U ~U ~U ~ ~- E tn ~ ~ f0 IO O N f0 f0 N cC f0 l0 f0 fG O O O D t O p p O
>,
N ~Q Q ~4 f9 N f9 f0 f0 Q L U U U U U U U U U U U U U U U U U U U U U U
O ~
m I
O O NI t~ n N a00
O O_ O N N 1~
00 ~ Z (O ~ N ~ Z U V
OOV~~IsO~Mpmtl~0 a00C00f00'~Otv~~~NMOM e-~GMOI~~f[~DIM O
(~D t! I N O O ~f ~_ N ~ ~ I N M O N fN~ I O ~I I ~I O V O M t~ N ~ Q7
0o w U o °' a> o N in U ,°'n ~ (~ ° N U ;° U M U o
w C7 ,~ ~ '_~ ~° U N m U
~ X ~ Y J D D ~ J ~ ~ ~ ~ J Z J ~ ~ ~ d.' ~ ~ ~ X Z cn ~ X ~ Z ~ ~ ~ X
~I ~I Q ~I ~I .-I ~I .-i 01 .-I O Z ml ~I ~I ~I .-I .-I .-I ~I .-I ~I .-I p .-
I
O~ N_ 01 a0 '? ~t CO f0 OW ~ O~ ~ O ~ O~ ~ M ~_ (O O~ (O M 1~ O CD M M h CD f0
CO c0 O_
a0 00 O ~ O ~ N M N (O Q) h ~f'7 (p O Q) 1n M sr C h a0 OD N N N ~ N (O N
1~ N V (D ~ V1 O V O '~! ~ ~ ~ N ~t M ~Y (D M (D 1l1 t0 m N 1~ ~1 1~ ~ O ~ st
1~ ~ O
M O 01 M M O M M N (D N O O ~ at tf ~ N M O V' V O O M
M M M O M O M M O O O O O O M O N M N N M O O O M O M O O O O N O
U ~ ~ ~ U U U N N ~ U ~ U U U N U ~ N N N U ~ N N U N U U ~ ~ U U
U U O U U U O U U U U O U O U U O O U O O O O O U U U O O O ~ U U O
lJJ L1J LJJ W tJJ UJ W L!J LIJ W W L!J lJJ W ~J w W W u1 w ~J W UJ UJ w llJ
lJJ U~ W L<J lJJ LIJ w UJ
103
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WO 01/11086 PCT/US00/22061
~'~bte 2, c~nt.
U
a~ U U
U U o 0
U <v N
Z Z
7 ~ - .~
t0
d d d t3. d Q d
H f- E-
~ f Z Z ~ Z Z Z Z Z Z Z Z Z Z Z Z Z
~ Z >- Z Z
Z ~
>-
Z
>-
Z
Z
Z
Z
Z
~
Z
Z Z
w.
L _ O - L N d
y O N
N d d d
N
Iw f-'
~
I- f- f- f-
Z >- Z ~ Z Z Z Z Z ~ Z Z ~ >- Z Z ~ Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z
Z Z Z 7- Z Z 7- Z Z Z 7- Z 7- ~ Z Z >- Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z
._ Y > N O
N v ~q '
C C 10 ~ C ~ O
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a 'a N a ~ a
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ca d d ~ C ~~ ~ ~ 7
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E (O In a N v ~ ~ ,C L ~ y ~O
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L_N_t0~- Q p_-- N C
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C C O a fO f0 N m O
d d d a ~ ~X O " N d ' C
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(n U) fn tn (n (n
U O O D 'o 'O ~ ~ '~ V N lL C~ N N N l1J UJ lil lL lil U! lJJ LLI Lll lL 111
liJ LiJ liJ UJ Lll 111 l1! lL
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t~ (O O 1n 10 f0 M M O) N N OD ~ (O O 01 M OD O
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Q V O O O O O N N N N N N M ~ ~f V
N = N ~ O ~ ~ h ~V sf ~ p V ~ O ~ M
o U '~ ~ ~ c°o U ~ U ~ ~ o o ~ ~i M U ~ U U U U U U U U U U U U U U U U
U
XI ~I XI NI ~I ~I ~I XI ~1 =I ~1 ~I ~I ~I ~I I ~I JI ~I ~I ~I ~I ~1 ~I ~I ~I
~I ~I ~I ~I ~I ~1 ~I ~I ~I
O ~ ~ ~ ~- Q ~ U ~ ~- Q D Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q
M O N M O '~! O_ N M h ~ h '~ M 00 ~f7 h CO ~ M O M h N_ (D ~ N
N (D Q~ Q~ 00 M f0 07 M M V 1n 1~ (O 01 f0 t~ O~ N O O O s_t 01 O (O 01 N Q1
(D ~
M N ~O CO N N N N O) 07 O ~ ~ a0 N M (O h t~ 1~ 00 N N M V O O M O
M ~f N M V V O N N N .- GO M '~t 'C ~ ~f '~f ~t .!7 In 10 ~ 1n ~ 1W O tf) ~ (D
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W W W W CL <L W W l1J LIJ LLJ UJ W W w 111 w W lJJ 111 w l1J W LL W U! W UJ UJ
W Ill UJ W W W
104
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
~'ab~e 2, c~n~
0
x
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Z Z Z Z >- Z Z Z Z ~ Z Z Z Z Z Z Z Z Z ~ Z Z Z Z Z Z Z Z Z Z Z Z Z
N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N
f-E-HHHHHHhf-HI""1-"HHf"f-E-H1"~f-HH~E"_~ f"'f-'f-_1-f-f-~f-
ln fn (n (n (n In (n (n In fn (n fn (n (n (n In fn (n fn In (n N fn fn (n (n
fn fn fn fn fn (n UJ fn In
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M V M O~ O (D h ~ M O (O t~ O f0 (D 01 O N M O 1~ O O ~ 00 ~
p~ tW N N O~ (D M N cO M N N a0 cD V m M N 00 I~ O~ N h M V' h O~ ~ O O~ 1W
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lJJ W 111 llJ
105
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
~'a~le 2, c~nt~
O U O U
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f- f- E- f- H- f- I-_ f- I-_ I-'y- ~ f- I- f" I-' f- f- f- ~ H H H H f- H I-
t"_ f- ~.. I- H H I- H"
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fn (n In fn fn (n (n In (n
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U U '-I U Z U
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lJJ w w W UJ W UJ LJJ lJJ V! l1J L1J W lJJ tJJ V! L1J LJJ LJJ L1J w W W W W w
111 lJJ lJJ 111 W W W lJJ l1J
106
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
'Table 2, oon~
0
x
U U
U
U
>. o
U X
Z
Z
_
(B
N N
1Z a
Z Z Z Z Z Z Z Z Z Z Z Z Z ~ Z
Z Z Z Z Z Z ~ Z Z Z Z Z Z Z
Z Z Z Z Z Z
U X
Z N
_ Z_
N
_ a_
a~ a~
a a
H H
Z Z Z Z Z Z z Z Z Z Z Z Z ~ Z Z Z Z Z Z Z 7- Z Z Z Z Z Z Z Z Z Z Z Z Z
z z z ~- z z z z z z z z z z z z r z z ~ z ~ z z r z z z ~ z z z z z r
M ao ~ ~ <o ~ ~ ~, E ~ ' cn cn cn cn cn cn "' z
°' o o Z ~ ~ t a ~ >- >- >- r >- ~ m d c ~ H z
O O J_ J_ J_ J_ J_ J_ J_ a
v v v Wn H
w m c U ~ ~ ~ ~ ~ ~ ~ ~ m a Q' ~
a> a~ a~ a~ a~ O z ~~ r tn Q Q Q Q Q Q Q 'm 'm N
m m m m m d z c ~°, ~ ~ I~ I~ u' t~ u' u' m c>o = O Q
g > > > > > > > o
m m m m m ~ ~ '~ .~ U ~ (n fn fn (n fn fn c c N m U
o » » » » ~ ~ ~ ~ Q g
O O O O O g ~ p Q J J J J J J J J C c C
c c c c c ~ ~ Q Q Q Q Q Q Q Q c 'c o ~, O
a~ a> u~ c~ u~ ~ -: _: -: -_: .: _: -: _: ~ c ~ Y a Q
c c c c c ~ Z o _. _. _. _. _. _. _. _. ~ a
O O O O O O O O O O O O O
(n .t.. > .:
m m c'a <'n m m c'a m m m c'n m p
0 0 0 0 0 0 0 0 0 ~E '~ ~E y y E 'E 'E E ~E ~E ~E ~E
f9 W f9 l9 _A _(O _IC W _(O ~N 'N 'N 'N 'N 'N 'N 'N 'N 'N 'N 'N 'N O
T >. T T 7, T T >, >, T T 7. T E
E E_ E E E E E_ E E ~ a~ a~ ~ a~ a~ ~ a~ a~ a~ ~ a~ a~ '
N N N N N N N N N O R ~ ~ ~ ~ ~ ~ ~ ~ O ~ tp T
T T T _T >. ?. >v >. T ~ N C7 d N O 47 N N d N N d Y
L L L L t L L L L 'p ~ ~ ~p ~ -p ~ ~ ~ ~ ~ ~ ~ (d
~ O O O O O O O O O O O O O d
FN H H H H F- FN IN H H EN H F- fN IN H H fN h H IN IN H F- h H fN EN H H ~ ~
EN IN H
tn N tn ~ (n (n (n (n (n (n (n (n (n tn In (n tn (n (n (n (n (n fn (n fn fn (n
tn (n !n (n In In fn tn
LL w W LL L!J W W lJJ LL 111 w LU 111 w lJJ L1J w UJ lJJ W W lJJ W W L!J UJ W
W lJJ UJ W LLJ LJJ UJ 111
D U U I Q U Q U ~-1 Q U
M ao ~ M_ m ao ~ o ~ cc r~ a~ a~ v ~n a~ o a~ ~ co I n
N
a~ v Wn I v t ~_ o~ n ca ao c M ao ca r~ ao w U o~ c~ I
p v1 N Q a~ N m N U Q m wn o~ o o ~ v ~_ o_ n ao M o r~ ~ M o o m
~ 1W M N N I 00 MI O 00 O O N f0 V 00 N V_ ~ ~ N O 1~ O ~I7 O) 07 t< D ~ ~'
Q_7 'vt
O O ~p N V I N ~ ~f O p N O (O I sf V' st tn O M st ~ O N st N ~ sf N tn N
v I 1 I I ~ I O I .- v I M I ,~ I I I I I I I I I I I I I I I op I I I N
d' ~ ~ d' Z ~ ~ d' ~ d' h ~ ~ ~ ~ d' ~ ~ ~ ~ ~ d' ~ ~ ~ ~ O ~
Q m m Q QI Q m Q QI Q Q Q Q Q QI Q Q QI Q QI ml 00 D U Q U D Q D Q Q Q ml Q Q
O Q~ O 07 O Q_~ m N M O ~t'f O N ~ a0 ~' ~ 00 4'! f0 N O_ h f~ O f0 M ~ OD fD
O_7
O h m M_ f~ C N 07 O O ~ st N h 07 O~ O st 00 O t0 O a0 O tn Q~ O~ ~ N V N_
01 O O O N V a0 N O~ O 00 ~ O 1~ M 1~ O st OD M (~ f~ 00 N ~ (D O N ~ ~(7 N O
N M M M M M M M ~ sf ~ th O~ (D t0 cD (D pp N ,-. p7 ~ 1~ N 11) M (D a0 ~ N ~1
V
M M M M M M M M M M M M M N M O O O O N N O N O N O N O O M
In fn fn fn fn (n fn In In fn (n fn N !n In UJ (/J fn UJ (n fn fn (n fn In V7
(n (n V7 fn fn (n fn fn (n
O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O
w l1! 111 W w W lJJ IJJ W w W L!J w lJJ lJJ W <L W lJJ U~ V! UJ W UJ L!J Ii!
lJJ W LL W LL W lJJ U! w
107
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, coat.
0
x
U
U U U T U U
..
U U U
U U
Z Z z
Z
Z Z
~
_ _ _(U _ _ _ _
m N m m N N
a d d d Q. a. a.
7. T ~ T >, >. >.
~
Z Z Z Z ~ >- Z Z Z Z Z Z Z Z >- Z Z Z ?- Z >-
~ Z Z Z Z Z Z Z
U
U U U
U U
Z Z Z
Z
Z Z
_ _ _ ~
~ _ _ _ _
N N N N N N N
n n n n n a a
>, T >, T >. ~,
f- H H I- f- H f-
z z z z ~ z r r z z z z z z z z z r z z z r z r r z z z z z
N
Z Z Z 7- Z Z r Z Z Z Z a Z Z Z Z Z Z Z Z Z Z Z Z Z Z ~ Z Z H Z
f9 '--'
'V O ~ ~ D O Z O _N ~U E N
N C N C O n ~ N N ~ M p~ n X
v ~ _p ~a~ m 'a~ y O ~ c_ Z L m d ~ c ca o
J ~ Z U E O N U O O N n E' fa C ~ N O
L Q n E O Z ~ E C_ c9 E U ~X U n
Z O O n O O 07 f0 O O O 7 7
N U 01 'D O n O ~ O) N a0 a0 C E N C N
f9 l1J E N T O d N O f9 V- U ~ ~ O N N O N C ~ C N
O O .L... O O ~ d a U p N N C O v L ~ X V O C
m c a E ~ o n a'~ :°_ v ,a a~ a~ 0 ~ ~ E °~ ~ c d
c ~ n m ~ ~ ~ a N o c . o N Y n p y ~ v ~ o~
~ a~ c = Q c ~ c p o' n ° m M ~' ~ ~ ~ c
p ~ ~ ~ ~ . c N ~ a N c ~ L o o ~ p o Q m
Q ~ ~ E N E ~ ~ '~ y U m o n c m N y ~ N cc n ~ c n p E U
N t4 O ~ n C ~ C O E
_ "7 E Z C O N. (n M N ~ w ~ C ~ '(p O t C O C N O O E w O
0 0 0 0 0 0 0 0 0 0 0 0 o y c ~ -- ~ c E o. n ~ ~ ~ Q ~, s
N N c4 fa f0 f0 ca fC l9 c0 f0 l0 t4 C C O ~ C C ~ ~ O N O ~ 'O" U ~ z C d O
~E E E E ~E ~E ~E ~E E E E E ~E ~ia ~ia °W n m ~ ~_ v 'a o ~ ~ ~
° ~°- ~ ~ a~ E
~fn ~N ~N ~N ~N ~fn ~t/1 ~Vl ~N ~N ~N ~N 4! N N ~ ~ ~ ~ J ~ O ~ C C p ~ Z Z E
M N
7. 7, 7, T T T T T T T 7. ~ >, vp t w ~ a T d d N ' N ~ O ~N
YYYYYY-YYYYYYY _~ _~ C_~a ny O p O O U IC E E
N O 'N N d N O N N N N N N U U L C C f0 .17 ~ U p ~ p U 7 N
~O ~O 'D ~~ ~ Q m ~ ~ ~C C O O C N G7 N N N ~
N N N N N fn N f/1 .fn N N t/1 N ~ ~ O C ~ C C O ~ O O O 'C N N fn0 N N fn4 t
I- F- ~ ~ F- ~ H ~ ~ I- F- I~ ~ Y Y T O E ~E n n N A N N N 1/! N N
!n (n (n ~ (n (n (n (n (n (n fn fn ~ > > ~ ~ ~ (9 f0 O f0 d ~ ~ 7 '~!
lJJ CJJ W 111 lJJ W w LGJ W W W V! W N d :~ w Ii. C ~.. w C7 O m C7 U' O _
D o n ~ n ~ ~ v t n UI U UI I
~'I O v N ~ N ~ ~ M ~ ~ N 1- ~ ~ ~i ~I M v
aOD MV M N O ~ ~ ~Cl ~V ~ ~ O _ = Q EI _ U U m
fN~ ~ ~ ~ f- ~ ~ ~ ~ ~ N ~ O ~ O O ~ f0 ~ N O M C~0 ~ O N H '? (D OOD
o U UI N U U U U U U U U U o M 'r N M M N N ip N c v n ~ M I I ~ ~ o
~ n. ~ °~ o C7 ~n m yn o m ~ in ~n ao U U p, o 0
I I I I I I I t I 1 I I 1 X ~ ~ X 2 X ~ J J X X X ~ ~ X X ~ ~ X X X
Q U Q Q Q Q m m D Q Q Q D .-I ~t .-I ~I ~i r-I .-I ~I ~I ~I ~I ~I ~t .-I ~I Q
DI D ~I ~I ~t
O~ (D ~ ~ N h 1~ f~ (O O M O O ~ M h M fD O N h N h N_ 07 ~ M_ ~ 00 'Q N CO CD
07 CO O_ O st c0 ~ t~ a0 st V N M N N a0 OD O ~ et M sf ~ t0 N O M O (O (O a0
O 00 (D 01 07 st ~ O CO a0 (D h M t0 O~ ID N_ h 00 O tn N M st N c0 M O N
N 1~ (D OD K1 N M N M N 1!7 (O M N N O N C~ ~ N N N M M In M M M N
M O N O M N M O O M O O O N O O O O M M O O O O M M N M O O M
(n fn (n fn (n (n (n fn fn N (n (n In (n (n In fn In N (n (I7 (n (n fn fn In
In In (n fn !n In In In fn
O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O
lJJ UJ w LJJ W (1J w l1J w lJJ LL! W W <L LL lJJ W lJJ LJJ LU W W IL LL W Lll
W W W W UJ W W UJ W
l
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, conk
o T
U U
U
U w. .r
j ~ Z Z
i z
Z
N f4
~ f>3 _ N
N N N ~ N N
d a Q. d d. Q.
f- F- E- H f- H
Z >- Z ]- Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z ~- Z
~ Z Z ~-
Z
U U
a~ Z Z
N c0
L ~ - f0 =
O' a T
~
> , > T >.
, . H H H
H N >
~ f- N
z r z ~ r z r z z z z z z z z z z z z z z z z z z z z z a~ r z r r
z z z ~ z z r r z z z z z r r z r r r r z z z z z z z r ° z z r z
N H U U U U U U ... >
Z O U
°~ c E ~ m m iu is m o 0 o E ~ ~ °- ,n Q ~ Q Z m
~ a' c r r r r r ~ '. .r Y a~ E ~ Y c Q Z ~-
x U N a~ m m m m m U n c d 'a~ Z ~ E H
w
U a~ a~ ~ hi c o a a a n n M M M J E y U o ~ E o
~ n c c c c c o 0 0 ~ c°~ o ° o. E "' n,
-- a~ a m ~ ~ ° 'a~ '~ 'a~ 'a, '~ ~ ~ ~ v N ~ o ~ Y E
M ~ y O tC W >. ~ ~ d a a d a U ~ ~ i(~7 tNC Z ~ L C V N ~O ,>J
O C U n C7 N G7 U
m Q Q Q a °' ~' ~ ~ v ~ a~°o m ~ r ~ ~ ~ 'c E .~ ~ o ~ ~ ~ o ~_
o Z 5i
N ~ ~ !r ~ v v ~ N- ° 0 0 0 0 ~ n p Q D m 'a~ ~ °' ~ ~ ~ a
~ o E E v>
t E E E ~ 'N ual ~° Y Q Q Q U ° Q. m y °-' a~ E oo d L a~
Q Y Y Y Y Y c Z Z Z Z s a ~ o ~ E . ~ U ,o"
Y ~ co ca t~ !r - Q Q Q m '- ~ ~ ~ ~ O
O Z V '° N N N E ~ y ~ " ~ ~ .~0. ~ w w U U U y ~ O ~ Q V c9 ~ Q ~
O
Q ° t o 0 o a Q Q Q Q Q Q Q Q Q Q Q 'N ~ ? L o Q :° ~ o ~ o
a
o ~ ~ N U U U U oWG E E E E E E E E E E E E .~ ~ c~ ~ ~ N a° ~ '-
° o :o
O f9 N N Vl V1 fn !A M N N f/7 N tA M t/) N V! N N f/f f6 C - O ~ E -L ~ Q
C ~ C C C C C C C C C C C C C C C C C C C > C '~ O M A O N O.
U y O VOf Q d d t1 d d d d d d d d d d d Q d d d ~ T 7, ~ O N EO O O t0/) d
~ X (9 N f0 f0 f9 f9 f0 la f9 f6 f0 (O f0 f9 lO l0 f0 fQ f0 f0 U U U C7 CJ L L
!!~ d ~' E E
L O C N M N N N N N N (A 4! In 4) 1I! N tIl fn 1/l N N C C C C C C C C C C C C
(n O -1C O O O O O O O O O O O O O O O O O O O cC ffl N fa l0 c9 cG N c0 la tG
f0
° E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E
y y y ~ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 > > > > > > > > > > > >
x r L L x x x x x x x x x x x x x x x x x x x x x x x x x x x x x x x
tn Q a~ Q Q 1 Q
I M O M M M Q I 00 CO w CO
O U O V N 1~ at V M O I
H' ~I J a00 000 O ~ ~''~ ~I (~O vc) OM0 (OD aMD
x Q Io v_so c_~~U oM.-m o v_
CO O O N Q7 CO O ~ O tW 07 M ~ N Z ~ ~I ~ O ~ ~ ~ ~ N N ~ 10f1 (M0 (~O
N O O t!7 M M I N o v I I o~ 1 1 r~ ~ I I N 1 ~ ~ ~ ~ ~ ~ ~ a I ao ~ t~ I
U o o n c~ N o~ U o o .p U U r~ U U r~ ao U U M U .p co ~ N .Q ca o U o co v U
x J X ~ ~ Z J ~ ~ Q ~ ~ ~ ~ ~ ~ Z p ~ ~ ~ ~ ~ ~ > > J J ~ ~ ~ ~ >
-I ~I Q ~I Q ~I ~I ~I ml Q Q Q mI Q ~I m m Z Q ~I ~I ~I ~I ~'I ~-1 <'I ~'I Q
~I ~I ~I Q
00 h ~f h O O V ~ N N (D N I~ CO tL1 t~ tn OD h O M 1~ tff CO ID O N t0 N tl)
to f~
00 O M t~ ~ i~ N m ~ ~ a0 O O M M O M V N tn 01 M O 01 (D M O ~ N 1~ N
O 00 t0 ~ 1~ V O~ ~ O 1~ Q7 (O f~ O~ ~ aD M O~ O X17 M M M ID O O_ N_ ~ st ~ O
M
O M O O O M M O O O M M N M M M M O ~ O N N O ~ O M O O O M O O O O M
C)
L!J W W lJJ UJ W W W W W l1J W l1J lJJ W LJJ LJJ lJJ W W W W LU liJ U! UJ W V!
w 111 UJ W t1J lJJ UJ
109
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, cont.
0
x
o a~
x U
a>
U >,
r.. U
U Z
Z
f0 (B ~ f0
Q d Q. d Q. d
f F- f- I-'
Z Z Z Z Z Z Z Z ]- ~ >- Z Z Z Z Z Z Z Z Z Z ~ Z
~ Z Z Z
Z
Z
Z
Z
U Z
l9
O (Q N - f9
a ma a a
~'H T H H
~
f E
F
r Z r ~ Z Z Z Z Z Z Z Z Z Z Z r r r Z Z Z Z Z Z Z Z Z Z r Z Z Z Z
z r r ~' z z z z z z z z z r r r r z r r z z z z z r r z r z r z z
c
O ~ N ~ ~ - - - _ ~ e4 tC
O O ~ X X X
O N N N N C L d d
U ~ L d (d L L L p ' O . p
N N 41 C C ~ ~ ~ O y O E O V1 V1
N O 'tn N N N > 7 > > a O O
U U a ~ T _f6 f' Q7 f0 !9 (C M tt L ~ t O D
c0 f0 m 01 m
r r o m E ° '_' °' a~ a~ a~ c_ c v m ~ ti ti
f9 c9 U ~ ~ ~ ~' m C C C OJ ~N ~..p. O C _N N
d d a N ~ L ~ C C C C N O O U N Y E N Q
l4 O N N td d d d w l0 f9
CC~'~U '~ C_C_C_C O1QI~N~ C CC
of o> E _" ~ '° Z m 'E 'E 'E y C C N N O 'o o a~ a~
C y C ~ ~ ~ V M N C_ C U U d ~ ~ n C f0 N
N .: p O _C ~ _O N N M ~p O L L, ~ p d N C_ ~ ~ 'N N GI
O7 m ~ 'O
O E ~ O U C 07 O O) L U p p E E ~ y N V O
G7 U d w '~ C C C ~ ~ V V C C 7 O N 'p0 '0 N N C d C C
U y U ~ N C ~ ~ 'D O f6 ~ O O ~ O C fC d
Y Y T v c v a~ o ° c c c L L w ~N ~N c Q a a~ ~ p c ~ o o N 'm 'm
~ CO N ~ O ~ '~ ~' Q Q Q N O ~ ~ L L O U °U O ~ p ~ ~E ~ l0 Y O (G
C O
Z Z U ~ m N ~' ~ D O D ~ U a~ oW° ~° _° d ° a~
o T r ~ ~ d d
~ ~ v i t ~ m a~ '.- v y a> a~ ~ ~ ~ c c m ~ ao m t -°' c ~' ~ ~ t L
E E ~ 'C .- .O d (p .,.. ~ O O O ~ J Y Y 7 7 CO Y Y C O O H ~ O ~ tn V ;O O O
C C C C C C C C ~ O O O p C C ~C O O N ~ y (A a C O N G1 C C y L
fn -N p (9 N f4 f4
7 7 7 7 7 7 O 7 E L L L ~O ~N ~f/1 ~N ~ d ~ r ~ C ~ ~ ~ C U E ~ 3 E N Q Q
v E c c c c c c c c c c c c ~ ~ y ~[ y N c9 J O T T ~
N ~ O OO O Q NI NI U
a0 ~ N ~ n UI N M 1~ N O_
f0 ~p M 1 ~ M Q' N O ~ NI h
I H I ~ N O f0
XI C XI = c0 Q = n7 t_O m a~ ~ ao
'Q M N O ~ 'p Of) et N ~V M O ~ (p M 00 ('~ M V M t~ N f0 N 1~ ~ O M N N N M
OD ~ 00 M f~ M = CO 1~ ~ N ~ CD ~ N M ~ (~O Or0 O CO OWD in ~ ~ 01 ~ O
~ N ~ ~ c0 ~T N C~ U QOf ~ ~ t( C~ M (O N M U N p U ~ N COO 00 U h U O U 000
nC1
O O ~ > > ~ > > Z ~ ~ ~ X X Z ~ ~ ~ ~ ~ D r ~ X X O ~ ~ tn ~ ~ ~ > > Q
~I D ~I ~I .-I ~I .-I ~I ~I ~I .-I Q ~I ~I U ~I .-I ~I ~I D .-I m ~I Q ~I ~I Q
N O (O M et 00 h OD tD fD OD t0 (O N h 00 (O O tn h 07 O O~ 1~ N Qf Q~ O O tn
at
N ~ 'C N ~ ~1' O ~ '? f0 ~ (D O 00 GO O h Q1 CO ~ N ~ M ~ (O 00 Q) (O N N M O
N
(D M M V V IO M f~ (O M 1~ O t0 1n ~ sf _~f O O ~ ~ OO N fD ~ O '? N a0 f~ V
O ~ N N N V O M O t~ M N O O N M N N N ~ N a0 C7 N N7 Q7 N N 00
M O M O O O O M O M N M O O O O M M O M M M M M O M O N M M M N O O N
~ N U U ~ N U U ~ N N N U
O p O O O O O U O O O O O O O O O O O U U U O U U U O U U O 0 U O O O
w lJJ W L1J W LlJ llJ l1.1 W LL lJJ l1J V! w W 111 W LL! W LL W UJ IL lJ.l IL
111 UJ W W W w w W V! W
110
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, coot.
0
V V
U U U L
U ~ Z
Z ~ Z ~ Z
_(U .D - .~
N N
!Z Q
T ~ a
T
Z Z Z Z ~ Z Z Z Z Z Z Z Z Z Z Z Z ~ ~ Z Z Z Z Z ~ Z Z Z
Z >- Z Z Z
x
d ~ z Z
Z
m a a a
T H H
H H 1
z z z z z r r z z z z z z z z z z z z r r z z z z z r z z z z z z r
z r r r z r r z z z z z z z z z z z r r z r z z z z z r z z z r r z
o c v _ v ~ o
O L ~ C CO O O 7
N ~ N C U D1 d
(9 N GI L O L j (NC ate.. C
C
y ~ ~V <U C N N
O O tU/f C _j d r N _ O
U NN ~ O N O V..~'. ~ dU
. _ N E p O T .a O x U O
p C
U.7, N UC LEC_ ~ O-Lp ON _ :YO NNO
p ~ ~ tpn m O p ~ ~p d ~ ~ N T N N ~ C E
E ~° ~- a E v ° o a ~ v ~, °' ~' ~ °? ~ ~c ~
a~
j< a O ~ ~ ~ N d N T ~ C v N (9 GO N C N
N N ' N C ~ N ~ U f0 0p O " (p ~ C'7 Q T c9
C ~ N U ~ (~ ~ ~ O N
N d :°_ v N > ~ m ayn v ? '~ ~v' y m
o .- ° ~ E '° o ~ c m L o a N c_ c ~ C~ v ~ d
p o o ~ a ~ . ~ v ~ m o °' Q, E c a ~ ~ . ~ .- o ~ x ._
N O w. 0I ~ ~ C X p p C
_=' (4 (9 C ~ O C L y L ~, ' a = y '_. O' E N O Q ~ ~ p N O v
C C O U . O O1 a V L d U C E p7 f6 N N ~ N ~ cC Q
C ~ 01 V
°.' :_' ~ _m E '~ - -'' c a~ E y X of c:i °' C > y ~ ~_ ~ N
C O O cD O y ' r d ~ E ~ O ~ C Q U .~C r ~ ~ ' C O p' C
O O O ~ 07 L ~ E O ~ O Z N O ~ '~ )' N loll f0/7 O O N y U N ~>' Ip
p ~ ~ Cp <6 U y ~ Of ~ ~ 3 O = 'U ~ E' fC f0 fC = ~ ~ C d U C O ~ C L
N O N N L E N U ~ U d L E ~ w C ~ _N C E ~O ~O O O t~ ~~ ' .~ O ~(C ~ O O
~ E E O N U ~ ~ C_ C C C C U f9 ~O O C ~X O L L L ~' C C ~ . d ~ _U ~0 !C
C C v f0 E O ~ ~ ~N ~N ~N O .. 7, N N ~ ~ N N N N .r0" E ~ O O E V .'- ~ N d
O
C N G7 O O O O O E U U U Y C ' O O O ~ N N W ' N p O O O O
f0 N fa i6 N V O >, >. T 7. T O U 7 7 C Q y y t L L . m O O O O
E E E E E E ~ E E E E E c c Z c c o a a a a o_ a a a a a a a a a a o_ n
I D NI (D
w ~ ~ wI O O 1"~ NI O
HI f~ f0 N O NI 00 NO
M O ' M ~ _I
O In O M V O (D ~ et N ~ M C'7 OI N (D fD 00 O O ~ ~ ~ (D G1 ~ Q1 M O I~ ~ (O
V M N N ~ O ~ ~ ~ O In Z U N ~ ~ ~ ~ ~ N sf ~ f0 ~ In ~ et ~ M t~ N O
ID C~ O~ a0 I 10 .~ N N M o7 I I O O O M I ~ ~ I ~t M I ~ I I (D OD O~
M IMn l~n o V ~ C7 0 't o V V ~o ago ~ ~ U ~ ch U °~ U o U m U f~ p'rp
N (h N o~
D X X X ~ ~ Z J ~ ~ '~ ~ ~ ~ D ~ ~ ~ D ~ D ~ ~ J ~ ~ ~ ~ ~ ~ ~ ~ J D
I (a r-I .-I ~I Q r-I ~I CJ O ~I ~I .-I ~I Q .-I ~I ~I Q Q ~I Q ~-I m Q Q
t~ N t0 ~ 00 O~ a0 O 10 (O ~f ~ 01 1f7 O h N h !'~ O ef 00 t~ c'~ h In N V ~
f0 c_~I O
h 10 (D ~ V M W ~ t~ In N m Q1 a0 CD 1f7 O N 1~ 1~ fD N h O 00 N 1~ (O N O t~
OJ f~
O Ip O~ 00 O (D 1~ O_ N O N O O C7 c_0 1l7 ~T O '.t O aO N 01 sf Q1 f~ M M In
O M O_ O f_~
O c~7 N N O~ N O O) N 10 sT at O 10 O O ~ 10 t~7 O O M h ~ N h
O c'~ O O N O O O N O M N P'7 O O O O O t~ O O O f~ f~ c~ O O O O ('~ O O fh O
(n fn In fn fn (n N (n In fn fn fn (n fn (n (n (n fn (n (n fn fn In fn fn (n
tn (n (n fn fn fn (n !n (n
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
w w w w w w w w w w w w w w w w w w w w w w w w w w w w w w w w w w w
111
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, coot
o
V ~ U
V U U
U U "
o o
> x
~ Z
o ~ .
- Z
Z -
Z
_
ca ca
c~ ~ -
~ -
a~
n ~ a
n
a . .
a
7, T >. >,
Z Z Z Z Z ~' Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z ~- Z Z Z Z
]- Z ~- ~ Z Z
U X
N Z Z N
O N
c4 L - ~
N
1" H
Z Z Z Z Z >' ~ Z Z Z Z Z Z Z Z Z Z Z Z ~ ~- Z Z Z Z Z ~ Z Z Z Z Z Z
z r r r z r ~- z z z z z z z z z z z r r z ~- z z z z z r z z z r r z
o c v ~ ~ o
O 19 t O C pp O 7
y .D ? ~ ~ N C
Q7 ~ N
U ~ ~ j U 47 =~ V ..~'. r T V d U
N W O) T L X O
N UC LEC ~ ~~ Ou1 - :Y~.0-00 tOn~O
N ~ T O m O p N ~w d O t/W' d N ~j C N E
N
U . E N N E ~ O O d ~ ~ ~, O T ~ ~ ~ .C L N
X N O' ~ ~ ~ N O d t/1 >. C v . _ N f9 W _O C f/1
d C ~ C C C ~ N U ;O ~ ~ O. U ~' (~ ~ ~ O y f9 ~ N N
_ ~ ~ B_ 7 ~T' ~~ t6
d O _ _
° " O O Q ~ L ~ O cC9 ~ Of ~ C a fE N ~ L C O N ~ X
_ f'
T IOn N N a N ~ U O. O " ~ f0 C d O E L O O ~ O E C
m m c ~ c r a~ L >. d Q 1° ai ° E hi o o ?? In c~ m
C C O Y ~ O 01 T V ~ ~ V ~ E O ~ ~ N ~ ~ N O ~ O N
'a~ 'ao ~ ~ E '~ ~ a~ E y X ~ ~ N c ' > ~ o ~ ~ ~o ~ c o
C OO O _c0 ° Y ~C _O d ~ E E p1 'O ' Q U ~C ~, V ~ d Q 01 'N N N
E O O O C O ~ ~ O ~ O Z N O C ~' ~, 10/1 N N O O N y U N T O O' C
p O. ~ f6 p ~ U ~ p1 N ~ ~ 3 U 7 ' ~ E _d _d ~ 7 p) ~ X O
~ N N t E N ~ ~ U d L E O ~ C W N C ~ ~O O O O M T ' ~° ~ c0 L' ~ N
r C7 E E o o ~, p v cc c c c '° ~ ° c ~X a~ L L L Z' ~ c ~ _c~
_~ m
p v v C c0 E C O N ~O ~O ~O ~ O E V ~U U Y C ~ O O O 7 ~ ~~ 'O ~ E T O
'W fC N i0 d V O T 7. T T 7. O U > > C Q O y ,= L L ."' O O O E O O O O
E E E E E E ~ E E E E E c c Z c c o a a a n a a a a a a a a a a a n. a
I O N M
v O .~1 O at~o M NI NI O
M H I M O C (O O ~.1 O 00 ~ =I
0 0 ~ wn ~ M
sT M N N O O O ~ N 01 ~ Z U N ~ ~ ~1 ~ ~ N '? ~ ~ (O W f7 ~ 'at ~ M h N O
W'rno~~Z~ ~~~a'~D~~~O~O~~J~°~d'»~~~JD
O x x X I 1 1 I I I 1 I 1 I I I 1 I 1 1 I I I I I 1 I I
~I ml ~I ~I ~I Q ~I .-I D D ~ ~ ~ ~ Q r- .- ~ Q Q ~ Q ~- m Q Q
M N cD In 00 01 a0 O In tD V ~ O ~ O h N 1~ M O st a0 I~ M h 10 N ~ ~ tD M O
h In (O V V M In ~ Im0 N 01 O 00 OD tn 01 N 1~ 1~ f0 N h O 00 N t~ 10 N 07 h
OO h
O Ip pi cp O~ t0 I~ O N O N O 01 M cD In V O s_t O 00 N O) V m ~ M O ~ O M O
O> 1_
O M N N 01 N O 01 N ~ ~ O Ih O C 117 M O O M 1~ ~ CV f~ 'Q'
O M O O N O O O N O M N ~ O O O O O M O ~ O M M M O O O O M O O M O
U U ~ ~ U ~ U U U N U U U ~ U U U U N U ~ ~ U ~ U U ~ N U U N
O O U O O O O O O O U O O O O O O O O O O O O U O O O O O O O U O O
W W LIJ LL! W LJJ w W V! W LlJ w w W LJJ tJJ w V! W W w l1J W Lll LL U~ W w W
W U~ w W LL W
112
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
'Table 2~ cont
0
r X
U U V
U U U U U
> U
z Z v Z
Z Z
N N
~ .17 f13 ~ ~ . -
d d
r E- f
~ Z Z Z Z Z Z Z Z Z Z Z Z >- Z Z >- Z Z Z Z >-
>- Z Z Z Z
Z Z Z Z ~
]-
X X ~ U
O O
N N Z Z ~p c0
Z Z
~ a ~ a ~ - _
a a Q n
H H H H H
H H
z z r r z z r z z z z z z r z z z z z z r z z r z z z z r z z z z
z r z r z r r z z r z z z z z z z r z z z z z r z z r z r z z z z
a~ a~ y ° T m o
c o 0
o E o~ ~ o E
E ~n L ° ~y c v ~ o L o
c _ o r. >. ° ° o. o, ° E
O O7 N V ° ~ L d
in d d ~ C tn ~ d ~ E E O M U E O Z
'.~ E N N.U N ~C ~ fir- ~ Q' C
U ~ V N ~ U N ~ C p_ T ~ C l9 C O L V O
E O O C N .C N w C O ~ f0 . p N Q7 N L f0 C . m p C
~ o. a rc ' o c .° Q o r- D v' v a~ ~ E a ~ E c °
O) N N d J ~ E L T ~ L C O 7 N O y ° 7 _f6 ~ ~>
v v ! (0 C C v O ~~ t v 100 L f0 U N C ~ ~ E d O
c . .~ o o n- m E aWo _a'~ o ~?? a~ ' >. ~ o~
E ~ ~ 'E C T = N ~ j ~ C V 'E E ~ ~ a ~ Y .E ,°'n ~ v o ~ °
E
L U a ~ C N O N p ° L O O O N T N f' ~
N .~ .o v d~ 't E E
L L Q ~ U O Y (~ C v O O N ~ O_ C O w U a O ~ L C y > l0 C
N N N ~ _O N - ~ ° y 01 C C N d y ~ T ~ > f0 O ' ~ O N f_6 1/! y
O O N E U ~y C ~ O T C U ~p ~ ~ C (0 E ~ L E (~ d X ~ N ~N O E t
r s m ~ L ~° D ~, ' °' ~ ° °.' 3 0 ~i ~' v o
°' ° a~ c~
N d ~ O ~ ~ d d V G) ~ ~ C N ~ C a Q O ~ m ~ C ~ ~ H ~ O C N N O
N C C p C O ~ ' a O C ~ -p d O C O O f' m -_ O
-- om, C E
N ~ ~ N d E ~ O_ ' Y O ~ V y " p C ~ O O ~ p C f0 L 'N E, C ~ ~ E ~ (C (/7
C t9 d ~ ~ 7 ~ ~ ~ ~ ~X O V U L ' C O E O M Y p O ' r
'° a ~ tn 0 v ~ ° °' 'x ° = c :°_ ~ ~ ~ a~
a~ ° ~ a~ aW 'n m E_
C C C ? O Y C ~ O [yJ C p ~ O t0 O ~ ~ ~ N O V ~ D1 Y E E C C ~ O ~ fn N m
01 (O l0 7 V C E Q O E L 41 (=/1 C C C C C d O ~O E N ~ ~ j~ N
d d d ' In Vl II) ~ f/7 Vl In N N N N r. r r .r. r. r..
aoi M N
1 v ~ ~ ~I m Q
ao ~ o ~ n co ~ NI m a~ M r- ~ n r~l n r~ ~ ,~ NI M rn
tn tp (O O O M O M ~ O 07 O O ~ ~ Q~ ~ 00 O 1l7 (O h t0 m
ca ooN Io~co,~NN~N~IajN~oN~m°M°~ I~p~NO~_a~oOMO~ Wn I
N ~ ~ D U ~ ~ ~ Z ~ N ~ U ~ X ~ X D J D ~ ~ O ~ ~ ~ ~ D X ~ ~ >
~I ~I ~1 ~I DI ~I ~I ~I ~I Q ~I ~I U ~I ~I Q ~I r1 ~I ~I m ~1 ~I ~I ~I ~I ~I
~I ~I ~I ~I ~I QI ~I ml
a0 ~ 00 ~_t'f N O~ ~ O O CO O~ O ~ '~1 00 ~t1 t0 M V' N Qf O ~_ ~f'f O 00 CO O
O u7
a0 V O M V M ~ O ~ ~ n t_D ~ 10 a0 O 10 t0 O O h h 01 h 00 O) N (D _~- ~ O) M
N
N 1~ ~ N M M ~ O) O fD N ~ O f_' O (O O (D N M
M O O Q) O N O a0 ~ M M M O M M M ~ M M M M O) M M
O M N O O O M M N M M M O O M M M O O O M M M O M N M M O
N (n fn CO In UJ (O (n N (n (O (O N In (n (O (n In (O fn N (n fn (O N fn (n (n
In (O (n In (n fn fO
C)
w W IL W W W w W W W w LL W W IL W w LL W W W W W w W W LL LJJ w W W w U~ w
11.1
113
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, conk
p U O U T O O O >
~ ~ ~ U
U
U X U ~ o U U U o
O > T O X 7, >, >, X
U z O U U U O
Z Z
Z ~ ~ Z ~ Z Z Z
_ _ _ _ _ _ _ _ _
_(~ ~
N N O N N N N N O O
O
o_ n. a a n_ a a a a n.
o_
f- f-- >' f-
~ ~ ~
y- ~ Z ~- ~ >- Z Z Z ~- Z Z Z >-
Z Z Z Z Z Z Z Z Z ~ Z Z Z
N > ~ N X ~. ~. >~
O U O
U z Z U U
Z ~ ~ Z ~ Z Z
._-_-_ ___- _m -_-_ ._
a_
a~ d a~ a~ a~ d d d a~ a~
a o. a a a d o. a a a
a
a
H H FT- H H H H H IT H
ET
z z z z z z z z r r r z r r ~ r z r ~ z z z ~- z z z z z r z
N z z r rz z z r r rz z ~ z z z z ~- z z z z zz z z z
z z z
c ~~o
O U d
d ~, E E a~
~ N
_ C O d
N 7 =
~Q ~
f4 O X
E ~ E ~ ~'
U
O a0f0
N 0- 3 C v E
C . .
= O .C7 O 41 N
O O
E Z ~ v N c0O
M
V _ ~11~ d ~ _ N 7 d
~ ~. N
~ ? c ~ D c v
N o a~ o ~'
n o y
~ a~ _a~ ~ ~ Q m n ~ m
N
OI C O N N ~ - O O >
O S'c
O N y C N C d ~ U C y
O
' U ~ E a''a~~ 'a~ ~ ~ a~m
~, '
> _ C O . Y
' 'vo c o ~ ~ L ~ . U
o
o~ u, i a '~a~ a ~o' ~ c c
E o o
C ~ i1 E ~ U L N E y E Z ~ O
O '- O
p7 C I o ~ _m a~ ~ p
O L U ~
m
ca r ~ L O C U ~ O O ~ ~ d
C U d X ~
~ O O N d j N ~ O __
~ O ~
(n ~ O r ~ O ~ O U ~ N
~ ~ V
fn Q ~ O N L N C " O C
CO E11f7 Cr C L j DN ~ f0C
f~j~ pp d~ C O C C CC y C ' CN CpN C1
N 01 O C
O
C N X E ~ u N VIN N (/7N N N f/1
0 ~ N
-
1-
c0 ~ O N C 'pU 7> GI~ C V1E ' ~ ~ F-H F-!-H F-F-F-F-h H F
O U fl)(l)(n(n(nfl)fl)!1)f/)
VJ
U C ~ >.E~ ~ O f0(C(4f4~ L O O >'fG~ C C (/J(l~fl)ILU!LJJ111111ILl1!liJUJ
N ~ U IL ILIlllJJ UJ
'
T N N N mm CD.oU U Uc U U U v C~N
>.
I
U
0
n N ~IO
~
N O
O ~Y N M ~ O 1t7f_DtDM_O)N ttC7
O Z O N O ~"~ V ~ O~n (D
1- (D
H M
I a0 n n MM n n O ~ ~ ~ OO n ~ p O ~ (OO ~ n O lf7n n ~ N
( ( M O n M O n O ~ Q) f0
N
N ~ ~~ O ~ a0p ~~ tn0 ~ ~0 l~l7t~~ '~MN
M X M ~ p C
~ O
_ ~ _ ~ ~
I I I l ao~ n n ~ o ~ M o Z ~ UZ
I I -
D O Z ~ ~Q X X J N J D Z> > J ~
f~ ~
o d0 O O OO (DM o tnM~ n 01COe-'~sf O n tnM~O~ 1 O NEOM O O~
n n O n
N of N 0OOa0 f0 n ~faON n (O O)t0O _ N (DQ)O Q7_ 00Q7O Q1f0t(7
CO f0 O
N N M (D~O~Dst4DN N 00sfO N n sfO O ~ O O N ODM (Dn n n N t0
~ n M ~ '?'vt~~ ~ ~ O
~
N
M V ~OM On M N M _ MM O ~ O ON O O M n N N~tO O O OO O O O
O M NN M O O MO M OO O O
M N N In(n(n(n(nIn(n(n(nfnInfnV7fn(nN InIn(nfnfnfn(n(nfnfnIn(nN
M O fn fn
In (n
(n fn
O O O O OO O O O O OO O O O OO O O O O O OO O O O OO O O O
O O O
UJ l1J V!W WLLW llJlJJW WW lJJW W UJW W UJW W w LllV!W L1JV!WW UJW lJJ
W LLI UJ
114
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, cont
O O O X
U U
(j U
V U U U
Z Z Z
N ~ N
d d d f1
>, >, T
Z Z Z Z Z Z Z Z Z ~- Z Z Z Z ~ Z Z ~ ~ Z Z Z Z Z Z Z Z Z
Z Z Z Z
U U U
N
d N N
p d Q d
, f- H H
z z z z z z z z z r z z z z z z z r z z r z >- z z z z z z z z z
z z z r z z z z z z z z r z z z z z z z r z z z z z z z z z z z
N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N
f- f- H E- ~ f- ~ ~ f- ~ f- H F- E- ~ H f- H f- H H" f- ~ ~ ~ ~ f ~ H ~ f- ~ ~
f" f-
W W W W W W W W W W W W W W W W W W W W ~ W W W W W ~ W W W W W W w W
1('! I~ Q) (O OD V O ~ ~ p~ M h O 000 CO 01 1n O
cD a0 O O~ 1~ N N aD O M O O
_ Of N ~ f0 N ~ N O~ N h ~ O h f~ OO V ~_ ~ N t~ O (O ~ tC~ M
WM17 ~ 0~0 n t0 N p n a0 M O~ M M u7 0~ (O M O ~ op u7 OD O~ M O h ID fD M 01
O~
N N M h N N tn M M Wn ~ GO (p ~1 1n ~C1 N ~ O M I~ m CO 1~ O M ~ ~ M 1~ ID
C 'ct 'Q V ~ O N p (_O m '~1 (O M N N N ~ '~t '? O M CO CO N (D O M 00 ~ N_ M
O~
LL Z Z Z ~ h- 0 ti Z I ~ ~ ~ ~ Z ~ ~ Q
pp ~ ~ N M ~ f0 t~ a0 t~ t0 M O M (~ ~ Q~ t~ 'Q Q_~ M M (D M tn M c0 O~ 01 N N
N
M ~(7 pp M N O QO M OJ O M M (D Q1 O O) N h V f~ V (D ~ r_' O CO X17 M O t~ O
00 O) N
O O O N h (O O ~"> M O CD O (O O O O O 1~ CO N ~ <D O O O V OW f7 ~C1 a0 ~f1
Q~ Q~
OOOOOOOOOpOO M V_u7_~_~_O_O_O_O_n_OONNNNNNNN
f~ N f~ f~ l~ t~ t~ f~ !~ f~ f~ N (~ l~ N f~ (~ f~ V7 (~ N N f~ N N N f~ (~ N
(~ f~ f~ (~ N f~
l1J W W W V! W UJ W 111 W 111 w 111 w 11.1 W w W W W w V! w W w UJ W W w W W w
U! W W
115
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, coot.
O U
U
U o
T X
U O
Z Z
N O
Q d
T
Z ~ Z Z Z ~ Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z
x
U O
a
C C
o. a °' °'
-a 'o.
m m
N N
N
.: .: C
_. .. O
U -'
Z
Q Q ~ Z
z r z z z z z z z r z z z z z z z z r ~ >- ~ z z >.. ° z z z r ~ z ~
z Z cn u~ ~ cn a ° p ~ '~ ~ _ ~ z Z
u. N . o z z
J J J J J ~ O S ~ ~a ~ ~p ~ ~ M "7
a c a Q Q Q Q Q --co ~ C a .H l1! '~ ~ ~ J_ J_
J N Q LL lL ll LL lL ~ = O C cC'4 (~
Q m w > > > > ~ a ~ ... Z ' In fn Q Q
_N ~ fn U) tn U) U) C ~ N ~ o U) ~ U7 In ~ lL
J J J J J C ~ O M L ~ ~ U U fn fn
M : .: .: .:
Q Q Q Q Q ~ U a~ ti Z F- Q Q Q Q
O _ . -. _. _. _.
U o 0 0 0 0 0 0 0 0 0 0 0
o uJ .: _: _: .:
vYtn'm'm~'aco'm'mcem''m~'oca'moo00
0 0 0 -~ -~ ~~ -~ ~~ -~ ~E y y
p ~N ~N ~N ~N ~N ~N ~N ~N ~N ~N ~N ~N ~ ~
7. T_T_>. T_>._>._T_T T Tj. E ~E ~E ~E
E E E ~ N N d 41 N N O N d N 1U ~N ~N ~f/1 N
N N N ~ f9 (C N (4 f' fC (C 10 f0 N f9 ~., >. T 7
7. 7. T y N G7 G7 d N N d d N N O Y Y Y -1C
t t t W ~ ~ ~ 'O 'O 'D ~ ~ ~ 'O ~ O !4 (D N
01 O~ OI O O O O O O O O O O O O d d O d
N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N
f- ~ f- f" f- f- f- ~ f- ~"" ~ f" ~- ~ ~ ~ H ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ f" f' ~ f"
N (n tn (n fn fn In In fn (n (n (n N fn (n tn In fn In (n (n (n (n (n In fn In
(n (n fn (n fn V7 (n
W W w w W W w W W W W w W W W w w W W W W W w w W W W w W W w W W W
O M M O M M M OO _M 00 O CO 01 O h M M M f~ '~f ~l1 ~D
M O ~c7 O~ 00 N M O ~f tn O O (D 00 ~ CO ~ ~ 00 V N M O
V ~t7 ~ ~ M N 1~ ~ et O 07 O a0 ~_ O_ h t0 N 07 C~ M h a0 tD M tn ~ O tD
OD ~ M tD c0 O_ O (D tn M N 07 f~ N N f0 M 00 00 t~ O OD N tn O ~ O O tD st ~
M 1n
N y11 tn V M M O ~ M 1~ O0 M tW 'p (D tn O t~ O CO O~ Q~ M_ CO ~ O ~ M CO O CD
M_
O O O N O 'Q ~ ~ ~ N 'at tt et ~ ~t O tn M O O (D '? ~O M M O '~t O O N (p O N
o ~ ~ ~ ~ o ~ ~ ~ ~ ~ ~ v ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ z ~ ~ z ~
O~ a0 N M n cD o ~ o M O M ~C) N (D N M o M fD f0 o M ~_ o U7 O tfi h tD G_O
(D ~ ~ O 1~ a0 a0 N O) M O) N OO O tn 00 M m of IO m O ~ h N ~ O1 O O ~ O
N N I~ W_l1 10 (O m O O N N 01 O ~f N M h a0 O ~ 1~ h. t0 a0 M (D 07 m ~ 1~ f0
a0
O O O N M M M ~ V 1f7 V_ ~ h 01 V ~ M M ~ 00 ~ M ~ N V ~ ~J '? N a0
N M M M M M M M M M M M M M M N N O N N N O O N N M N N M O
~ N ~ ~ ~ ~ ~ ~ N N N ~ N ~ N ~ N N ~ ~ U
O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O
W V! W W U! W LJJ W IJJ W W W W 111 W W tJJ w W W LL W w W W W UJ LL W 111 UJ
W W UJ LJJ
116
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, coast
0
o X
x °'
U
U
U
U Z
Z
_f0
d
H
Z Z Z Z >- Z Z ~ Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z ?~ Z Z
U
U Z
_fC
N
a a
H
N
_d
Z Z Z Z >- Z Z ~ Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z a> Z Z Z Z 7- Z Z
d
.o E
O O
U
N N
Q7 U
Q. C C
N N
d
z z z r z z z z r z ~ z z z r z z z z z r r r ~ z >- ~~ z r z z r z r
O U U ~ 4f N Z T N
M N Q ~ O E ~ ~ ~ N
a ago U Q '° ~ c%~ c c ~ r r E o ~ z Y E Q o v d
M M ~ C O r ~ ~, r U N N O O y D ~' N Z H N N
a~ a~ ~ °~ c m c v~ o o a a a a c C7 C7 ~ v v ~ t a~ a>
.a .D ~ E C 'U ~x O d ~ C_ C_ C_ ~ E O a E U U U C a N N
f0 f4 Q d p C ~ y M d G7 N N N ~ O V ~ (/) C f0 f9 ~9 E ~ y
y y yo E ", c ~ ~ x ~ E o 0 0 ~ ~ ~' ~ cc z 'a~ r r r ~ o ° a
N f~D = d ~ C U ~ ~ ~ ~ C V d d d Q Q ' ~ C E a d d j U T E
O O = w j 'V7 ~ C C E C7 O Q (D O) ~ N ~ ~ ~ O ~ U O N 41 N U Z T U
C C ~ ~ ~G ..O- O E T Z sf f~ Q1 C l6 T U C C C O U
a~ a~ p c N ~a ~ M ~.°? o E o ~o ~ ° o o ~ a ~ E ~Y a o~ ?~ m a~
a~ c ~ m Q
C C ~ p ~ N N 'P_ O d C .C p E O N ~ O 01 O Q> Q1 p E N Z
C C N d Q ~p ~ ~ ~ Y Q f4 N N v OO (D O M 01 C p
~ ctn'oQL ~v ~.n ,~'n'-°~YYY cZp o c ~ aVooo ~'o p
O O O O O .a d p C C Z ~ E d ~ N w w .Ø. w w U N O d ..p.. ~ tn d V p
°3 ~ :°_ m o E
C c0 N m
m m m m m o 'o °~ a E Q Q Q Q Q Q u~
_E~ E d E o~~~~~~ ~, ~ N jYYY-a
'In 'V1 N ~4) 'N ~ d O C O Y E T T ~ U E E E E E E N ~3 C C -~ w ~ '~O- O
T 7. T T T ~ ~ N H d V ~ ~ ~ V O C C C C C C C C A O- - t4 ~p Z Z Z Z U ~ d
m m a - - - - -m m ~ v c Q > > Z a, "- a> > a~ a~ a~ a~ a~ a~ a~ a~ a a a o ~
~ ~ ~ ~ t d
a> a~ a~ a~ a~ ~ c_ ° Z c Z m m ~ 'o ~ ~ ~~ m m m m m m m C7 r !~ E E E
E ~ a a
~ G7 C ~ y ~ O ' ~ 1n N N N M M N ~1 C C C C C C C C C C C
(D p C ~C Q ~p. N ~ ~ O O O O O O O O f9 l0 (6 ffl (0 f0 N !4 cC N f0
H >. ,°n 'a v ~ c N N L E E E E E E E E E E E E E E E E E E E E E
(O (O (O In CO ~ Y cc ?~ o ~ v o 0 0 0 0 0 0 0 0 0 > > > > ~ > > ~ >
IL uJ IL LU uJ :~ ti v~ m o> C~ Z S Z t Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z Z
Z
M
O
O ~ M ~ O
N f~ n t!7 M (O Q~ ~ ~ sf M O O t0 ~ ~ ~ f0 N ~ h M M O OO O 07 f0 M M Q1 ~ n
01
O aD p~ (O ~ M ~ aD 00 ~ ~1 c0 M 1~ py0 tW 00 N (O O O O OD O t0 M 00 N N ~ ~
O
O ~ O M M a0 ~ O ~ M _ O 07 V N f'~ M Q~ sf st O n O M M ~ O ~ f00 ~ ~ of a0
N (O N O C '~ N O~ O O O h M O I~ tt (O V M
O Q~ M V7 O M C7 f0 (D fD ~ I~ 00 tn CO N O O tn '? 00 m O N O N
~ X > > _ ~ X X X Z J ~ Z ~ D Z ~ N.' ~ J ~ O D ~ Q X
Q> O c0 M 1~ 1~ 1~ 1~ M_ ~_ ~f O CO 00 (D ~ t0 00 M O a0 N N f~ a0 f0 CO O f~
1~ t0 'Q
(D ~ f0 O M a0 M ~ ~ O ~_ a0 00 M N 1~ 07 ~O M ~ O N 1~ O) N s_T
N V M M O t_~ ~ ~ ~ N ~ O N O O~ ~ Q~ a0 M M O> O) ~ M O_ M ~ ,~ O N N M O N
~p ~p v(7 t(7 M M M N O V fn N M O M 00 N f0 O O_ O ~
N O N O M M M O O O O M O M N M O O M O O O M O M O M O ~ O O
N N N N ~ ~ ~ N ~ ~ N ~ ~ ~ ~ ~ ~ N N N
W L1J LlJ W W W U~ w IlJ W lL l1J lJJ L1J W !JJ 111 L1J W UJ lJJ W lJJ w W W
UJ LLI W w UJ W llJ UJ LL
117
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
'Table ~ coal.
0
x
U U
U
U U
0 0 >,
X X U
Z
Z
_
_ N
_(9 .a ~_ _(0 _.
~ N N N
d a d d a
T ?. ~, >. >,
f- E- H ~- f-
Z Z Z ?- Z Z Z Z Z Z Z Z ~- ]- Z Z
Z Z Z Z Z Z Z Z Z Z Z Z Z
Z ~- ~- Z Z
X X U
Z
Z
_
_ f0
_m a_ a_ _m -_
a~ a~ d a~ a~
n n n n n
T T T 7. T
H H H H H
z Z z r z z z z z z z z z z z z z z z z z r r z z z z r r z z z z z
r z z r r z r z r r z z r z z r r z z z r z z z r z z r z r z z r z
w
t L m N ~ N N L
n a ,~ in a 0 ~ o ~ c~ m c
o. v°m°ny~a)'o~c_~c°''
'O ~9 O E O N J U V ~ C U O N
C L O D ~ N U - 7 U - ~ O T
ch O UUVN tpn a'v_V t VV p_N
'O U '- O ~ - N
N N ~ T C d 4=. O ~O -O ~ 7
n n - E T 'Y O d O y d n E ~ f0
N C N O - C ~ U ~.' L' C
_y E ' ~ . . . p Q
E C E p N C n n _C v u0- O 7, d U V YO ~O N p N
V y C ~ ~ ~ C U U ~ Or C n ~ ~ ~ N ~ ~ L ~ C O
U N d N ~M L O- f0 L L ~ O d m
O) ~ ~ 'D N N ' U ~ ~ ~ C C
n O C ' C O C N n M of t4 N .. ' d m O 'O O, N C r
0I O ~ V d C_ C_ C C ~d .C C 'Y 01 ~7 ~ ~O ~ N ~p ~ N
L ~ N N O n O p N ~O ~p O ~O ~ ~C O) 10/1 N ~ > > A N '~ _fp
G7 C 10 L U O' N n V E c9 Y p p O l9 m ~ N Q L ~ . p C
17 ~ d ~ C ~ O ~ _ N n n U (0 - y U ~1 N~ p Q ~N N C U
p ~ (p ~p L _Y Vl d
t m ~' d O L ~ ~ O ~ N p~ ~ ~ y ~ O f0 N N O C C O ~ ~ ~ d N
c Z _n ~ ~ ~C O) ~ n O O y N L L _f0 ~c °: a a~ a~ c° a~ a~ v ~
~ c c ~ m
j ~ O = C ~ N U 19 a C ~ ~ O O O ~ E ~ ~ ' ~ Q O D) Of C X N N O ~ C
O O N T O 01 '~ O
E .Y L ~ N v U ~ E E
C ~ ~ = O O N C C C O X E E ~ ~ f6 N N C C C
f0 ~ ' _v n d U ~ c0
i. o o . °-' c°j E ~E °'
c ~E ~E ~ E m Q Q m m a~ i. ~ ~ r r ~ m m m m o 'o o ~0 0 0
~ C7 c c c ,~ Y ;~ m m ~ T >. g ~ E E E E c n n a a a a o. a n n a n n a
rn
M
H_
_ ~ O (O Cn0 _ 0~0 t~D
N ~ a0 tt> M tD N 'Q O tp O op c0 'p N O N N N N Of c0 ~ ~ (O M ~ ~ ~!7 O N n
t(~ O tn
O U Ot~7 O N N O ~ O 00 O t~~7 N m m COO st ~ M O off ~ p O. I' at M ~ ~ f~ '?
of t' O M
Z ~ X ~ O > j ~ ~ ti > > j X X ~ D ~ ~ ~ ~ r X ~ ~ ~ ~ ~ ~ X Q
0o m o ~ M o o~ o~ v o v v v <o cmn M o n M v v M n o w_ _n v o M
O st 01 N O) 1~ Q1 a0 aO (D O N M O (O t0 ~ ~ N t0 1~ N N (D N a0 (O O 01 O M
M (O O a0 _~ O ~ M (O O~ (D st op h V f0 pW O M (O O s_t e- CO O Q~ 07 s_! 07
M O~ O O O~ N
O O N M N N N M N O N N N N N N M ~f N N O M I~ ~ N M N N
N O ~ O O M O M O M O M N O O M O O M M M O O O M O O O M O O M O M O
~ ~ U ~ ~ ~ U ~ N U ~ U U N U N N U N ~ N N ~ ~ U
O O O O O O U O O U U U O O O U O U U O O O O O O O O O O U O O O O O
W LIJ W tlJ W w w UJ V! W LIJ W V! tJJ lJJ l1! V! W W W w W U~ W W lJJ V! W IL
W w W LJJ llJ w
118
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
Table 2, cont
0
x o
U
~, U
> U w'
Z U
~ Z
_(0 _ _(0 _ _
N ~ N N
Q. d d a. Q
T T T T T
~
Z Z Z Z ]- Z Z Z Z Z Z ~ Z Z Z Z Z
Z Z Z ~ ~ Z Z Z Z Z Z Z
> U O
Z r.
O
Z
m m -
! - ! . _ E _m
M '
>. T T >. T
f- N
H H H H ~1
Z Z Z Z ~ Z Z Z Z Z Z Z Z ~ Z Z Z Z Z ~ Z ~ ~ Z Z Z ? Z Z w Z Z
Z Z Z ~ >- Z Z Z Z 7- ~ Z Z ~ Z ~ Z ~ Z ~ Z ~ Z Z Z Z N Z Z '~ Z Z
D> O U Z Z C f' f' G7 ~ N N U N O
o Q '° vi o m '-> ~. ~. a 'a ~ c c p E
0 o E v v ~ Q Q ~ N ~a n °' o Z
~ L O Q O d d C >~ ~ O E E O O N (/~ N
N DON-L-' ~rQd E X00 =OOE~M
r ~ :~ tn In ~ a~ m o ai ai a c L L o o Z = = Z m
p c O t11 IL x ~ c " c c m ~ c N ~ m a a~
E U ~ Z Z ~ c °~ .n ~ m m ~ ° ~ ~ M t~ r~ Z = ~ U
i0 c C ~ ~ ~ N E p O N N C ~ d O O O ~ M N Q
y O 7 N ~ C T N O O L p O U U # ~ ~ C Z N d
'O (n m C T ~ O v U U U U C C C f' L
Q O j, ~ ~ O ~ ~ E ~ C m m y N m O O C CI O) a w j O
~C L Y U 'D N Q a ~ C L ~ C O .O m m D1 E O L N fR 7 p C C - ~
U - U m ~ N N y U E ~ E E . ~ C ~ C
o ' ~ ' c c ~ ~ a ° E o ~ 0 0 ~ c o > ~> m a~ ;~ m a~
c .~ ~ ~ m - ~ Y ~C C O ~ ~ ~ 7 d ' n. m ~ E o E m m ~ ~ ~ c
y N m ~ m o 0 0 ~~_ a 3 N m s_ o L L o ~ y ~ E 'E a~ ~ a~ N N N a~
O C ~> C O L E U U ~~ ~ C w U U ~ U 3 V d _d N ~ L .Ø. Y Y ' ~ m ~ ~ ~ N
c C7 N . ~ O_ O y p m m O U C7 d m m C ~ m > > ~ ~ m m m
~U O N ..p. C O ~ ~ ~ O C C N w ' V-. N d m ~ m 'V N 47 (n _ O O O N
d U Q ~ O T U U W L Q O 0 ~ L ~ C N ~' d N O ~ N C C ~ ~ U N N In
m ., ~ ~ ~ v v o m o ~ ~o. n. o N " Z ~ m °'. E ~ ~ m
M O ~ O C ~ N ~ ~ a m ~cj .i~ E _c c ~ Z Z V ~ ~ a~ ~~ y
E U O '
Q U U a~ O) 0 m m ~' C E N ~. .. ~" _ ~E N a a V in N in f~ f' N N M O ~ O (p
a w v a~ ~ E E ~ T ~ N m m m ~ yn O O c d m a~ U a~ a~ 0) !o M N
v (n N N N fn N N (/J N '~ > > > 7 7 > > > > T T >, T >v T N
O
1~ V O ~p M O ~ O~ O f0 V ~ O~ ~ a c0 ~ 0~7 N ~ O N O N O 00 ~ 1n V' aO N O
O_~
N f~ M '~ M OQ a0 ~ O M sf O~ 'vt M O OQ p O .p N sf OQ O O Q h st O CO
D ~ ~ ~ ~ Z H X ~ X D ~ 2 ~ ~ X ~ IM- Z Z ~ ~ ~ Z ~ F~- F~-- ice- ~
(O ~ ~ OD 117 N a0 07 (D M OD (p In 01 07 1f1 4'1 M 1n ~ M ~ CO O ~ O tn O tW
l7 e1 O
O) N O) '~ '~ O ~ a~ O a0 O 1~ 00 00 M N 01 N O) ~_ W et 07 N N OD O) O tn CO
O ~f ~ M _~ O_) c0 ~ M M h 1n O~ (D ~ 1_~ f~ O V' N 01 e1 O_O N O 1W V f~
O O M O N 00 m ~ N ~ M ~f 07 M f0 1~ O 1~ O) ~ ~f M M tn O 1n 01 01 M
O M M N O O M M O M M O N M N O N O M N M N M N M N O
(~ f~ N N (~ l~ l~ (~ (~ !~ (~ f~ (~ (~ f~ (f7 N f~ N (~ f~ !~ N N f~ !~ N N N
N f~ f~ N (~
O O O U O O O U O U U O O O O O O O O O O O O O O O O O O O O O O O
lJJ tJJ V! !1J LL lJJ W W W W W L!J W UJ w LJJ W lJJ l1J tL UJ W l1.1 W w W W
W tJJ W UJ W W UJ
119
CA 02381699 2002-02-07
WO 01/11086 PCT/US00/22061
TABLE 3
Exemplar
Accession Com lete Title UniGeneID
11/29/99
D86425 Homo sa iens mRNA for nido en-2 Hs.82733
D86983 Human mRNA for KIAA0230 ene; artial Hs.118893
cds
HG1098-HT1098C statin D
HG1103-HT1103"Guanine Nucleotide-Bindin Protein
Ral, Ras-Onco ene
HG3342-HT3519Id 1
J03764 lasmino en activator inhibitor; t Hs.82085
a I
L06797 chemokine C-X-C motif ; rece for 4 Hs.89414
fusin
L15388 "Human G rotein-cou led rece for kinaseHs.211569
GRK5 mRNA,
phosphodiesterase 4B; cAMP-specific
L20971 (dunce Hs.188
Droso hila -homolo hos hodiesterase
E4
L35545 endothelial cell rotein C/activated Hs.82353
rotein C rece for
L76380 calcitonin rece tor-like Hs.152175
M21305 Human al ha satellite and satellite Hs.247946
3 'unction DNA se uence
M24736 selectin E endothelial adhesion moleculeHs.89546
1
M31166 entaxin-related ene; ra idl induced Hs.2050
b IL-1 beta
M31551 lasmino en activator inhibitor; t Hs.75716
a II ar inine-ser in
M32334 intercellular adhesion molecule 2 Hs.83733
2 0 M61916 laminin; beta 1 Hs.82124
M68874 "Human hos hatid Icholine 2-ac Ih
drolase cPLA2 mRNA,
M74719 transcri tion factor 4 Hs.75356
M92934 connective tissue rowth factor Hs.75511
M94856 fatt acid bindin rotein 5 soriasis-associatedHs.153179
2 5 003057 sin ed Droso hila -like sea urchin Hs.118400
fascin homolo like
003877 EGF-containin fibulin-like extracellularHs.76224
matrix rotein 1
018300 dama e-s ecific DNA bindin rotein Hs.77602
2 48kD
027109 Human re romultimerin mRNA; com lete Hs.32934
cds
031384 uanine nucleotide bindin rotein 11 Hs.83381
3 0 033053 rotein kinase C-like 1 Hs.2499
059423 MAD mothers a ainst deca enta 1e ic; Hs.79067
Droso hila homolo 1
070322 ka o herin im ortin beta 2 Hs.168075
081607 kinase scaffold rotein ravin Hs.788
083463 s ndecan bindin rotein s ntenin Hs.8180
3 5 089942 I s I oxidase-like 2 Hs.83354
X04729 Human mRNA for lasmino en activator
inhibitor t a 1
X06256 inte rin; al ha 5 fibronectin rece Hs.149609
tor; al ha of a tide
X07820 matrix metallo roteinase 10 stromel Hs.2258
sin 2
X54925 matrix metallo roteinase 1 interstitialHs.83169
colla enase
4 0 X54936 lacental rowth factor; vascular endothelialHs.2894
rowth factor-related
X60957 t rosine kinase with immuno lobulin Hs.78824
and a idermal rowth factor
X67235 hemato oieticall ex ressed homeobox Hs.118651
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
X67951 roliferation-associated ene A naturalHs.180909
killer-enhancin factor A
X69910 H.sa iens 63 mRNA for transmembrane Hs.74368
rotein
X79981 cadherin 5; VE-cadherin vascular a Hs.76206
ithelium
218951 caveolin 1; caveolae rotein; 22kD Hs.247266
"zp61 b6.r1 Stratagene endothelial
AA187101 cell 937223 Homo sapiens
cDNA clone IMAGE:624659 5', mRNA se
uence"
N24990 ESTs Hs.26418
881003 Homo sa iens serine rotease mRNA; Hs.154737
com lete cds
AA025351 ESTs Hs.134797
AA027168 ESTs Hs.10031
AA040465 ESTs Hs.8728
AA045136 ESTs Hs.22575
AA054087 hos holi ase A2; rou IVC c osolic; Hs.18858
calcium-inde endent
AA071089 ESTs; Moderatel similar to !!!! ALU Hs.187932
SUBFAMILY SC WARNING
AA085918 H.sa iens HUNK/ mRNA Hs.247482
AA187490 ESTs Hs.21941
AA227926 ESTs Hs.6682
AA234743 ESTs Hs.22120
AA236559 ESTs; Weakl similar to neuronal threadHs.8768
rotein AD7c-NTP
AA292694 ESTs Hs.3807
2 0 AA398243 ESTs; Moderatel similar to defline Hs.21806
not available 3694664
AA406363 ESTs Hs.30822
AA411465 ESTs Hs.8619
AA412284 oliovirus rece for Hs.171844
AA423987 ESTs Hs.7567
2 5 AA425309 ESTs Hs.33287
AA435896 ESTs Hs.18397
AA448238 Homo sa iens mRNA for KIAA0915 rotein;Hs.16714
com lete cds
AA478778 ESTs Hs.16450
AA621714 ESTs Hs.25338
3 0 D51069 Human isolate JuSo MUC18 I co rotein Hs.211579
mRNA 3' variant ;
UDP-N-acetyl-alpha-D-galactosamine:polypeptide
T34527 N-acet I alactosamin Itransferase Hs.80120
1 GaINAc-T1
097519 odocal xin-like Hs.16426
AA127221 ESTs Hs.71059
ESTs; Moderately similar to C-1-TETRAHYDROFOLATE
AA132983 SYNTHASE; CYTOPLASMIC H.sa lens Hs.44155
3 5 AA135606 ESTs; Weakl similar to !!!! ALU SUBFAMILYHs.189384
SB WARNING
AA156125 ESTs Hs.72116
AA179845 RAB6 interactin ; kinesin-like rabkinesin6Hs.73625
AA232645 ESTs Hs.42699
F10399 ESTs Hs.14763
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
H16772 ESTs Hs.31444
N39584 ESTs Hs.17404
UDP-N-acetyl-alpha-D-galactosamine:polypeptide
N52006 N-acet I alactosamin Itransferase 1 Hs.80120
GaINAc-T1
N53375 Homer; neuronal immediate earl ene; Hs.166146
3
N54067 Homo sa iens mRNA for NIK; artial cds Hs.3628
N64436 ESTs Hs.20813
826892 ESTs Hs.221434
T33637 ESTs Hs.6841
"yc20g11.s1 Stratagene lung (#937210)
T57112 Homo sapiens cDNA
clone IMAGE:81284 3', mRNA se uence."
W80763 ESTs; Moderatel similar to FK506-bindinHs.3849
rotein 65kD
AA046808 ESTs; Hi hl similar to 40S RIBOSOMAL Hs.108957
PROTEIN S27
AA253217 ESTs Hs.41271
AA255991 ESTs Hs.175319
AA258138 ESTs Hs.88297
AA426573 ESTs Hs.41135
AA443793 ESTs Hs.94761
AA490588 ESTs Hs.43118
AA496257 ESTs; Weakl similar to defline not Hs.72165
available 3513303
AA609717 ESTs; Weakl similar to MICROTUBULE-ASSOCIATEDHs.66048
2 0 D59570 ESTs Hs.17132
F13787 ESTs Hs.58596
H88157 ESTs Hs.41105
H98988 ESTs Hs.42612
N34287 unc5 C.ele ans homolo C Hs.44553
2 5 N52090 EST Hs.47420
N66845 ESTs; Weakl similar to !!!! ALU CLASS Hs.165411
B WARNING ENTRY !!!!
N68905 small inducible c okine A5 RANTES
832894 ESTs Hs.45514
861715 ESTs Hs.138237
"yi54c08.s1 Soares placenta Nb2HP Homo
sapiens cDNA clone
IMAGE:143054 3' similar to gb~M87908~HUMALNE32
0 71234 Human
carcinoma cell-derived Alu RNA transcript,
(rRNA); gb:S41458
ROD CGMP-SPECIFIC 3',5'-CYCLIC PHOSPHODIESTERASE
"yr30g11.s1 Soares fetal liver spleen
898105 1 NFLS Homo sapiens cDNA
clone IMAGE:206852 3', mRNA se uence."
T97186 small inducible c okine A5 RANTES
W80814 ESTs; Moderatel similar to !!!! ALU Hs.193700
SUBFAMILY SB WARNING
AA404418 EST Hs.144953
3 5 AA405747 ESTs; Moderatel similar to HMG-box Hs.97865
transcri tion factor
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Accession Com lete Title UniGeneID
11/29/99
AA488687 ESTs; Moderatel similar to !!!! ALU Hs.190307
SUBFAMILY SQ WARNING
AA599143 ESTs; Moderatel similar to !!!! ALU
SUBFAMILY SQ WARNING
AA608588 ESTs Hs.193634
AA608751 ESTs; Moderatel similar to !!!! ALU Hs.244904
SUBFAMILY SC WARNING
C13961 EST Hs.210115
D60302 ESTs Hs.108977
H94892 v-ral simian leukemia viral onto ene Hs.6906
homolo A ras related
N93521 transcri tion factor 4 Hs.241362
N95477 ESTs Hs.102943
860044 ESTs; Weakl similar to !!!! ALU SUBFAMILYHs.106706
J WARNING
870506 ESTs; Moderatel similar to transformation-relatedHs.107159
rotein
"ye20f05.s1 Stratagene lung (#937210)
Homo sapiens cDNA
91518 clone IMAGE:118305 3' similar to contains
Alu repetitive
element;contains MER12 re etitive
element ;, mRNA se uence."
T95333 ESTs; Weakl similar to Strabismus Hs.122730
D.melano aster
845630 ESTs; Hi hl similar to KIAA0372 H.sa Hs.170098
iens
"yg05c07.r1 Soares infant brain 1NIB
820839 Homo sapiens cDNA clone
IMAGE:31444 5', mRNA se uence."
823858 ESTs; Moderatel similar to envelo Hs.23986
a rotein H.sa lens
A1024874 ESTs; Weakl similar to defline not Hs.57958
available 3882257
W26247 U5 snRNP-s ecific rotein 220 kD ; Hs.6413
ortholo of S. cerevisiae
AA856990 ESTs Hs.125058
2 0 AA136653 ESTs
AA358869 ESTs; Hi hl similar to SEC13-RELATED Hs.227949
PROTEIN H.sa lens
AI123976 ESTs Hs.105689
AI369384 a Isulfatase D
AA379500 ESTs Hs.193155
2 5 849693 ESTs Hs.107708
AA195678 Homo sa iens mRNA for KIAA0465 rotein;Hs.108258
artial cds
M30257 vascular cell adhesion molecule 1 Hs.109225
AA028131 ESTs Hs.110342
M10321 "Human von Willebrand factor mRNA, Hs.110802
3' end"
3 0 J03040 secreted rotein; acidic; c steine-richHs.111779
osteonectin
M86933 amelo enin Y chromosome Hs.1238
AA012933 tubulin-s ecific cha erone d Hs.241687
AA286710 m hoc a ada for rotein Hs.13131
I
AA243278 ribosomal rotein; mitochondrial; L12 Hs.109059
3 5 D59711 ESTs Hs.237289
" ye36g7.s1 Stratagene lung (#93721
T94452 I ) Homo sapiens cDNA clone Hs.241207
MAGE:119868 3', mRNA se uence"
123
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA053400 ESTs Hs.241227
AA370302 Homo sa iens mRNA; cDNA DKFZ 58611518 Hs.21739
from clone
J05008 endothelin 1 Hs.2271
U85193 nuclear factor I/B Hs.33287
AA256153 ESTs Hs.23912
X83107 BMX non-rece for t rosine kinase Hs.27372
AA046593 ESTs Hs.28959
AA410480 ESTs Hs.30089
D45304 ESTs Hs.31595
M90657 transmembrane 4 su erfamil member 1 Hs.3337
AA010163 a stream re ulato element bindin roteinHs.3383
1
AA136353 ESTs Hs.38022
Y07867 irin Hs.38842
U84573 rocolla en-I sine; 2-oxo lutarate 5-dioxHs.41270
enase I sine
X60486 H4 histone famil ; member G Hs.46423
AA132969 metallo rotease 1 itril sin famil Hs.4812
AA114250 KIAA0512 ene roduct Hs.48924
F13782 LIM bindin domain 2 Hs.4980
AA283035 ESTs; Weakl similar to !!!! ALU SUBFAMILYHs.54813
J WARNING
2 0 AB002301 Human mRNA for KIAA0303 ene; artial Hs.54985
cds
AA056731 S'o ren s ndrome anti en A2 60kD; ribonucleoHs.554
rotein
U68019 MAD mothers a ainst deca enta 1e ic; Hs.211578
Droso hila homolo 3
H99198 ESTs; Moderatel similar to THYMOSIN Hs.56145
BETA-4 H.sa iens
AA598702 bone mor ho enetic rotein 6 Hs.6101
2 5 N77151 Homo sa iens mRNA for KIAA0799 rotein;Hs.61638
artial cds
AA505133 ESTs Hs.62273
AB000584 rostate differentiation factor Hs.116577
D12763 interleukin 1 rece tor-like 1 Hs.66
AA253193 ESTs Hs.6631
3 0 AA432248 ESTs Hs.6738
AA083572 v-ral simian leukemia viral onto ene Hs.6906
homolo A ras related
AA479713 ESTs Hs.71962
L40395 Homo sa iens clone 23689 mRNA; tom Hs.170001
lete cds
X52947 a 'unction rotein; al ha 1; 43kD connexinHs.74471
43
3 5 W80846 vesicle-associated membrane rotein Hs.74669
5 m obrevin
M34539 FK506-bindin rotein 1A l2kD Hs.752
D67029 SEC14 S. cerevisiae -like Hs.75232
U09587 I c I-tRNA s nthetase Hs.75280
M85289 "Human he aran sulfate roteo I can Hs.211573
HSPG2 mRNA, tom lete
4 0 D10522 m risto lated alanine-rich rotein kinaseHs.75607
C substrate MARCKS;
W84712 calumenin Hs.7753
D29992 tissue factor athwa inhibitor 2 Hs.78045
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Exemplar
Accession Com lete Title UniGeneID
11129/99
L34657 latelet/endothelial cell adhesion Hs.78146
molecule CD31 anti en
S78569 laminin; al ha 4 Hs.78672
D43636 Human mRNA for KIAA0096 ene; artial Hs.79025
cds
U97188 IGF-II mRNA-bindin rotein 3 Hs.79440
AA487558 ESTs Hs.8135
M28882 "Human MUC18 I co rotein mRNA, com Hs.211579
lete cds"
X70683 SRY sex determinin re ion Y -box 4 Hs.83484
X14787 thrombos ondin 1 Hs.87409
AA236324 ESTs; Weakl similar to !!!! ALU CLASSHs.92381
A WARNING ENTRY !!!!
C15324 ESTs Hs.93668
AA452000 ESTs Hs.94030
D83174 colla en-bindin rotein 2 colli en Hs.9930
2
D00596 Homo sa iens ene for th mid late s Hs.196351
nthase; exons 1; 2; 3; 4; 5;
D11428 eri heral m elfin rotein 22 Hs.103724
D13640 ma'or histocom atibilit com lex; classHs.183618
I; C
D14874 adrenomedullin Hs.394
D26129 ribonuclease; RNase A famil ; 1 ancreaticHs.78224
D28476 th roid hormone rece for interactor Hs.138617
12
D86425 Homo sa iens mRNA for nido en-2 Hs.82733
2 0 D86983 Human mRNA for KIAA0230 ene; artial Hs.118893
cds
D87953 N-m c downstream re ulated Hs.75789
HG1862-HT1897Calmodulin T a I
HG2614-HT2710"Colla en, T a Viii, AI ha 1"
HG2639-HT2735Sin le-Stranded Dna-Bindin Protein
Mss -1
2 5 HG2855-HT2995"Heat Shock Protein, 70 Kda Gb:Y00371
"
HG3044-HT3742"Fibronectin, Alt. S lice 1"
HG3342-HT3519Id1
HG3543-HT3739Insulin-Like Growth Factor 2
HG4069-HT4339Monoc a Chemotactic Protein 1
3 0 HG417-HT417Cathe sin B
J03764 lasmino en activator inhibitor; t Hs.82085
a I
L06797 chemokine C-X-C motif ; rece for 4 Hs.89414
fusin
L08246 m eloid cell leukemia se uence 1 BCL2-relatedHs.86386
L12711 transketolase Wernicke-Korsakoff s Hs.89643
ndrome
3 5 L13977 rol Icarbox a tidase an iotensinase Hs.75693
C
L15388 "Human G rotein-cou led rece for kinase
GRK5 mRNA,
L19871 activatin transcri tion factor 3 Hs.460
L20859 Human leukemia virus rece for 1 GLVR1Hs.78452
mRNA; com lete cds
L42176 four and a half LIM domains 2 Hs.8302
4 0 L49169 Human GOS3 mRNA; com fete cds Hs.75678
L76380 calcitonin rece tor-like Hs.152175
M15990 v- es-1 Yama uchi sarcoma viral onco Hs.194148
ene homolo 1
M23254 ~ calpain; large polypeptide L2 Hs.76288
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Accession Com lete Title UniGeneID
11/29/99
M24736 selectin E endothelial adhesion moleculeHs.89546
1
M26576 colla en; t a IV; al ha 1 Hs.119129
M27396 as ara ine s nthetase Hs.75692
M31166 entaxin-related ene; ra idl induced Hs.2050
b IL-1 beta
M31994 "Homo sa iens aldeh de deh dro enase
ALDH1 ene, exon 13
M32334 intercellular adhesion molecule 2 Hs.83733
M35878 insulin-like rowth factor bindin roteinHs.77326
3
M36429 ostmeiotic se re ation increased 2-likeHs.89672
12
M57730 a hrin-A1 Hs.1624
M57731 GR02 onco ene Hs.75765
M60858 nucleolin Hs.79110
M62994 filamin B; beta actin-bindin rotein-278Hs.81008
M68874 "Human hos hatid Icholine 2-ac Ih drolase
cPLA2 mRNA,
M69043 nuclear factor of ka a Ii ht of a tideHs.81328
ene enhancer in B-cells
M74719 transcri tion factor 4 Hs.75356
M75126 hexokinase 1 Hs.118625
CD59 antigen p18-20 (antigen identified
M84349 by monoclonal antibodies Hs.119663
16.3A5; EJ16; EJ30; EL32 and 6344
M92843 zinc fin er rotein homolo ous to Zf Hs.198309
-36 in mouse
M92934 connective tissue rowth factor Hs.75511
2 0 M93056 rotease inhibitor 2 anti-elastase ; Hs.183583
monoc e/neutro hil
M94856 fatt acid bindin rotein 5 soriasis-associatedHs.153179
M95787 traps elfin Hs.75777
S76965 Protein kinase inhibitor human; neuroblastomaHs.75209
cell line
S81914 DIFFERENTIATION-DEPENDENT GENE 2 Hs.76095
2 5 003057 sin ed Droso hila -like sea urchin Hs.118400
fascin homolo like
003100 catenin cadherin-associated rotein Hs.178452
; al ha 1 102kD
003877 EGF-containin fibulin-like extracellularHs.76224
matrix rotein 1
008021 nicotinamide N-meth Itransferase Hs.76669
014391 m osin IC Hs.82251
3 0 031384 uanine nucleotide bindin rotein 11 Hs.83381
032944 d nein; c o lasmic; Ii ht of a tide Hs.5120
040369 "Human s ermidine/s ermine N1-acet
Itransferase SSAT ene,
041767 "Human metar idin recursor mRNA, com
lete cds"
048959 Homo sa iens m osin Ii ht chain kinaseHs.75950
MLCK mRNA;
3 5 051010 "Human nicotinamide N-meth Itransferase
ene, exon 1 and 5'
051478 ATPase; Na+/K+ traps ortin ; beta 3 Hs.76941
0l a tide
Human ovarian cancer downregulated
053445 myosin heavy chain Hs.15432
homolo Doc1 mRNA; com lete cds
059289 cadherin 13; H-cadherin heart Hs.63984
059423 MAD mothers a ainst deca enta 1e ic; Hs.79067
Droso hila homolo 1
4 0 062015 "Homo sa iens C r61 mRNA, com lete
cds"
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Accession Com lete Title UniGeneID
11/29/99
063825 Human he atitis delta anti en interactinHs.66713
rotein A di A mRNA;
067963 Human I so hos holi ase homolo HU-K5 Hs.6721
mRNA; com lete
073379 Human c clin-selective ubi uitin carrierHs.93002
rotein mRNA; com lete
073824 euka otic translation initiation factorHs.183684
4 amma; 2
077604 microsomal lutathione S-transferase Hs.81874
2
081607 kinase scaffold rotein ravin Hs.788
089942 I s I oxidase-like 2 Hs.83354
X04412 elsolin am loidosis; Finnish t a Hs.80562
X06985 heme ox enase dec clin 1 Hs.75967
X07820 matrix metallo roteinase 10 stromel Hs.2258
sin 2
X12876 keratin 18 Hs.65114
X15729 DEAD/H As -Glu-Ala-As /His box of a Hs.76053
tide 5 RNA helicase;
X52541 earl rowth res onse 1 Hs.738
X53416 filamin A; al ha actin-bindin rotein-280Hs.76279
X54489 GR01 onco ene melanoma rowth stimulatinHs.789
activit ; al ha
X54925 matrix metallo roteinase 1 interstitialHs.83169
colla enase
X57206 inositol 1;4;5-tris hos hate 3-kinase Hs.78877
B
X59798 c clin D1 PRAD1: arath roid adenomatosisHs.82932
1
X60957 t rosine kinase with immuno lobulin Hs.78824
and a idermal rowth factor
2 0 X65965 H.sa iens SOD-2 ene for man anese su
eroxide dismutase
X69111 inhibitor of DNA bindin 3; dominant Hs.76884
ne ative helix-loo -helix
X70940 euka otic translation elon ation factorHs.2642
1 al ha 2
X87838 catenin cadherin-associated rotein Hs.171271
; beta 1 88kD
X91247 thioredoxin reductase 1 Hs.13046
2 5 X97748 H.sa lens PTX3 ene romotor re ion
Y00815 rotein t rosine hos hatase; rece for Hs.75216
t e; F
AA303711 a hrin-B1 Hs.144700
L44538 ESTs Hs.156044
AA025351 ESTs Hs.134797
3 0 AA027050 ESTs Hs.31189
AA029462 ESTs Hs.17235
AA045136 ESTs Hs.22575
AA047437 ESTs Hs.22968
AA054087 hos holi ase A2; rou IVC c osolic; Hs.18858
calcium-inde endent
3 5 AA071089 ESTs; Moderatel similar to !!!! ALU Hs.187932
SUBFAMILY SC WARNING
AA156450 ESTs; Weakl similar to Similar to Rat Hs.8982
tr ene roduct
AA187490 ESTs Hs.21941
ESTs; Moderately similar to PROBABLE
AA195031 G PROTEIN-COUPLED Hs.9305
RECEPTOR APJ H.sa iens
AA205724 ESTs Hs.10119
4 0 AA227926 ESTs Hs.6682
~
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Accession Com lete Title UniGeneID
11/29/99
AA227986 ESTs Hs.25329
AA234743 ESTs Hs.22120
AA253216 ESTs Hs.22283
AA256210 oncomodulin Hs.199134
AA256268 ESTs Hs.10283
AA279397 ESTs; Moderatel similar to fibronectinHs.25001
H.sa iens
AA292379 ESTs; Moderatel similar to !!!! ALU Hs.20340
SUBFAMILY SQ WARNING
AA292717 ESTs; Weakl similar to JM2 H.sa iens Hs.7891
AA346551 ESTs Hs.23457
AA400292 ESTs Hs.23786
AA404338 ESTs Hs.21812
AA412284 oliovirus rece for Hs.171844
AA423987 ESTs Hs.7567
AA428594 ESTs Hs.21321
AA430108 ESTs Hs.6019
AA431462 ESTs Hs.28329
ESTs; Weakly similar to CAMP-DEPENDENT
AA431470 PROTEIN KINASE Hs.3407
INHIBITOR; MUSCLE/BRAIN FORM H.sa
iens
AA443756 ESTs; Moderatel similar to defline Hs.6673
not available 4105275
AA449479 ESTs; Hi hl similar to defline not Hs.5216
available 5106787
2 0 AA459916 brad kinin rece for B2 Hs.25021
AA465226 ESTs Hs.28631
AA478778 ESTs Hs.16450
AA479037 ESTs Hs.7961
AA482597 ESTs; Hi hl similar to defline not Hs.26054
available 4704739
AA487561 ESTs; Hi hl similar to RAS-RELATED Hs.9813
PROTEIN RAB-1A
AA489245 ESTs; Weakl similar to s erm s ecificHs.5682
rotein H.sa iens
AA504110 ESTs Hs.18063
ESTs; Highly similar to SERINE/THREONINE
AA520989 PROTEIN Hs.9195
PHOSPHATASE PP1-BETA CATALYTIC SUBUNIT
H.sa iens
AA599434 ESTs Hs.25035
3 0 AA608649 Homo sa iens clone 23742 mRNA; artialHs.6354
cds
AA609519 ESTs Hs.26458
D51069 Human isolate JuSo MUC18 I co rotein Hs.185718
mRNA 3' variant ;
U97519 odocal xin-like Hs.16426
W28391 roliferation-associated 2G4; 38kD Hs.5181
3 5 AA035638 Homo sa iens mRNA; cDNA DKFZ 564F053 Hs.71968
from clone
AA083514 ESTs Hs.68301
AA121315 ESTs Hs.70823
AA147186 ESTs Hs.92387
AA156125 ESTs Hs.72116
4 0 AA188932 ESTs Hs.85640
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Accession Com lete Title UniGeneID
11/29/99
AA219653 ESTs Hs.87125
AA232645 ESTs Hs.42699
F10078 ESTs Hs.13233
H48032 ESTs Hs.9645
H82117 ESTs Hs.28043
N39584 ESTs Hs.17404
N54067 Homo sa iens mRNA for NIK; artial Hs.3628
cds
N59858 ESTs Hs.33032
N90933 ESTs Hs.4867
N93764 ESTs; Moderatel similar to !!!! ALU Hs.10175
CLASS C WARNING ENTRY
826124 ESTs Hs.24024
827957 ESTs Hs.24230
855470 ESTs; Moderatel similar to K02E10.2 Hs.11067
C.ele ans
T16550 ESTs; Hi hl similar to vacuolar roteinHs.6650
sortin homolo h-v s45
T26674 ESTs; Weakl similar to neuronal threadHs.6966
rotein AD7c-NTP
"yc20g11.s1 Stratagene lung (#937210)
T57112 Homo sapiens cDNA Hs.8881
clone IMAGE:81284 3', mRNA se uence."
T88700 ESTs Hs.173374
T90527 ESTs Hs.7890
W42789 ESTs Hs.31446
2 0 W60002 lastin 3 T isoform Hs.4114
W78175 ESTs Hs.17901
W84768 ESTs Hs.141742
W94427 ESTs; Weakl similar to Na;K-ATPase Hs.3807
amma subunit
AA253217 ESTs Hs.41271
2 5 AA426573 ESTs Hs.41135
AA432374 ESTs Hs.48029
AA446622 ESTs Hs.74313
AA478771 ESTs Hs.50841
AA482594 ESTs Hs.62684
3 0 AA490588 ESTs Hs.43118
D59570 ESTs Hs.17132
H88157 ESTs Hs.41105
H94648 ESTs Hs.41995
H97538 ESTs Hs.42392
3 5 H98670 ESTs; Weakl similar to defline not Hs.49753
available 4884081
N22107 ESTs; Moderatel similar to !!!! ALU Hs.172241
SUBFAMILY SC WARNING
W38197 Accession not listed in Genbank
W80814 ESTs; Moderatel similar to !!!! ALU Hs.196785
SUBFAMILY SB WARNING
AA287347 ESTs Hs.105088
4 0 AA402799 ESTs Hs.182538
129
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA404418 EST Hs.144953
AA425107 ESTs Hs.97016
AA425435 ESTs; Moderatel similar to !!!! ALU Hs.98438
SUBFAMILY J WARNING
AA442872 ESTs Hs.110771
AA452860 ESTs; Moderatel similar to !!!! ALU Hs.197214
SUBFAMILY SP WARNING
AA488687 ESTs; Moderatel similar to !!!! ALU Hs.190307
SUBFAMILY SO WARNING
AA599674 ESTs; Weakl similar to ORF D.melano Hs.108115
aster
F13673 ESTs Hs.99769
H99093 DEAD/H As -Glu-Ala-As /His box of a Hs.6179
tide 72kD
"yw35g11.s1 Morton Fetal Cochlea Homo
N22495 sapiens cDNA clone Hs.102415
IMAGE:254276 3', mRNA se uence."
N23031 m osin; heav of a tide 7; cardiac muscle;Hs.929
beta
815740 carboh drate chondroitin 6/keratan Hs.104576
sulfotransferase 1
839610 cal ain; lar a of a tide L2 Hs.76288 .
W45560 ESTs Hs.102541
239833 H.sa iens mRNA for Rho6 rotein Hs.124940
240583 ESTs Hs.101259
AA825437 ESTs
866613 Homo sa iens mRNA; cDNA DKFZ 564F053
from clone
AA868063 carboh drate chondroitin 6/keratan
sulfotransferase 1
"z116dO8.r1 Soares_pregnant_uterus_NbHPU
2 0 AA128075 Homo sapiens
cDNA clone IMAGE:502095 5', mRNA se
uence."
N66570 ESTs
A1051390 ESTs
AA627122 ESTs
X02761 fibronectin 1 Hs.118162
2 5 AF010193 MAD mothers a ainst deca enta 1e ic; Hs.100602
Droso hila homolo 7
AA149044 ESTs; Hi hl similar to the KIAA0195 Hs.10086
ene is ex ressed
082108 solute carrier famil 9 sodium/h dro Hs.101813
en exchan er ; isoform 3
D78676 ESTs; Moderatel similar to defline Hs.105509
not available 4529890
L35240 eni ma LIM domain rotein Hs.102948
3 0 AA598737 lactate deh dro enase B Hs.180414
869417 ESTs Hs.107055
ESTs; Weakly similar to Human pre-mRNA
AA232837 cleavage factor I 68 Hs.107125
kDa subunit H.sa iens
N72695 ESTs Hs.108557
M30257 vascular cell adhesion molecule 1 Hs.109225
inhibitor of DNA binding 2; dominant
3 5 M96843 negative helix-loop-helix Hs.109617
rotein
X68277 dual s ecificit hos hatase 1 Hs.171695
AA292440 m eloid differentiation rima res onse Hs.110571
J 03040 secreted rotein; acidic; c steine-richHs.111779
osteonectin
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA228107 ESTs Hs.54642
AA449789 connective tissue rowth factor Hs.75511
W01367 ESTs Hs.170980
AA610116 ESTs; Hi hl similar to defline not Hs.11663
available 4325180
AA258308 Homo sa iens mRNA; cDNA DKFZ 564F053 Hs.165618
from clone
AA460273 Homo sa iens mRNA for KIAA0517 rotein;Hs.12372
artial cds
AA286710 I m hoc a ada for rotein Hs.13131
T68873 metallothionein 1 L Hs.143289
D63476 PAK-interactin exchan a factor beta Hs.172813
M62403 insulin-like rowth factor-bindin roteinHs.1516
4
X55740 5' nucleotidase CD73 Hs.153952
L10284 calnexin Hs.155560
AA243278 ribosomal rotein; mitochondrial; L12 Hs.109059
AA430032 ituita tumor-transformin 1 Hs.159626
H16402 ESTs Hs.17121
D59711 ESTs Hs.17132
"ye36g7.s1 Stratagene lung (#93721
T94452 ) Homo sapiens cDNA clone
IMAGE:119868 3', mRNA se uence"
AA431571 ESTs Hs.17894
879356 Homo sa iens mRNA for KIAA0544 rotein;Hs.19280
artial cds
2 0 AA280375 ESTs Hs.19928
249269 small inducible c okine subfamil A Hs.20144
C s-C s ; member 14
241740 ESTs Hs.24462
AA121543 Homo sa iens mRNA for KIAA0758 rotein;Hs.22039
artial cds
J05008 endothelin 1 Hs.2271
2 5 AA101878 ESTs Hs.22793
ESTs; Highly similar to (defline not
T35341 available 4519883) Hs.22880
H.sa iens
N87590 ESTs Hs.23037
AA256153 ESTs Hs.23912
W74533 Homo sa iens mRNA for KIAA0786 rotein;Hs.24212
artial cds
3 0 U25997 stanniocalcin Hs.25590
V01512 v-fos FBJ murine osteosarcoma viral Hs.25647
onto ene homolo
V01512 v-fos FBJ murine osteosarcoma viral Hs.25647
onto ene homolo
V01512 v-fos FBJ murine osteosarcoma viral Hs.25647
onto ene homolo
V01512 v-fos FBJ murine osteosarcoma viral Hs.25647
onto ene homolo
3 5 X56681 ' un D roto-onto ene Hs.2780
AA161292 nterteron; al ha-inducible rotein 27 Hs.2867
i
AA491465 ESTs Hs.28792
AA046593 ESTs Hs.28959
D50914 Human mRNA for KIAA0124 ene; artial Hs.30736
cds
4 0 D45304 ESTs Hs.31595
M90657 t ransmembrane 4 su erfamil member 1 Hs.3337
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Exemplar
Accession Complete Title UniGeneID
11/29/99
W69127 _ Hs.3449
ESTs; Weakl similar to zinc fin er
rotein ZNF191 H.sa iens
AA316186 ESTs; Hi hl similar to defline not Hs.34549
available 4262136
AA384503 ESTs Hs.179260
AA136353 ESTs Hs.38022
AA044755 ESTs; Weakl similar to !!!! ALU SUBFAMILYHs.173705
SX WARNING
procollagen-lysine; 2-oxoglutarate
U84573 5-dioxygenase (lysine Hs.41270
h drox lase 2
AA058911 ESTs; Weakl similar to membrane I Hs.4193
co rotein M.musculus
AA620962 d nein; c o lasmic; Ii ht intermediateHs.44251
of a tide 2
AA285290 small EDRK-rich factor 2 Hs.44499
X60486 H4 histone famil ; member G Hs.46423
831641 ESTs Hs.197148
AA489190 ESTs Hs.48320
F13782 LIM bindin domain 2 Hs.4980
AA257993 Janus kinase 1 a rotein t rosine kinaseHs.50651
M24283 intercellular adhesion molecule 1 Hs.168383
CD54 ; human rhinovirus
ESTs; Weakly similar to PIM-1 PROTO-ONCOGENE
AA443114 SERINE/THREONINE-PROTEIN KINASE H.sa Hs.5326
lens
T35289 casein kinase 1; al ha 1 Hs.195206
N23817 Homo sa iens clone 23675 mRNA se uenceHs.5807
AA047151 ESTs Hs.5897
2 0 N77151 Homo sa iens mRNA for KIAA0799 rotein;Hs.61638
artial cds
AA480074 ESTs Hs.62206
Y00787 interleukin 8 Hs.624
T99789 ESTs Hs.64313
W84341 tissue inhibitor of metallo roteinaseHs.6441
2
2 5 L09209 am loid beta A4 recursor-like rotein Hs.64797
2
D12763 interleukin 1 rece tor-like 1 Hs.66
T16484 ESTs Hs.6607
AA253193 ESTs Hs.6631
AA432248 ESTs Hs.6738
3 0 X82200 stimulated traps-actin factor 50 kDa Hs.68054
AA083572 v-ral simian leukemia viral onco ene Hs.6906
homolo A ras related
L00352 low densit Ii o rotein rece for familialHs.181182
h ercholesterolemia
N75791 ESTs Hs.7153
X57579 H.sa iens activin beta-A subunit exon
2
3 5 X02612 c ochrome P450; subfamil I aromatic Hs.72912
com ound-inducible ;
H44631 i mmediate earl rotein Hs.737
AA090257 su eroxide dismutase 2; mitochondrialHs.177781
X83703 H.sa iens mRNA for c okine inducible Hs.74019
nuclear rotein
L40395 Homo sa iens clone 23689 mRNA; com Hs.170001
lete cds
4 0 AA227913 ESTs Hs.198456
132
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
X52947 a 'unction rotein; al ha 1; 43kD connexinHs.74471
43
M11313 al ha-2-macro lobulin Hs.74561
L14837 ti ht 'unction rotein 1 zona occludensHs.74614
1
M60721 "Human homeobox ene, com lete cds"
D90209 activatin transcri tion factor 4 tax-resHs.181243
onsive enhancer element
"yc28e12.s1 Stratagene liver (#937224)
T67986 Homo sapiens cDNA Hs.75106
clone IMAGE:82030 3' similar to b:X14723
CLUSTERIN
AA148318 Human mRNA for KIAA0069 ene; artial Hs.75249
cds
097105 dih dro rimidinase-like 2 Hs.173381
T25747 H.sa lens OZF mRNA Hs.75471
K02574 Accession not listed in Genbank
tyrosine 3-monooxygenase/tryptophan
D78577 5-monooxygenase Hs.75544
activation rotein; eta of a tide
X53331 matrix Gla rotein Hs.75742
S73591 a re ulated b 1;25-dih drox itamin Hs.179526
D-3
X95735 z xin Hs.75873
L16862 G rotein-cou led rece for kinase 6 Hs.76297
044975 Homo sa iens Kru el-like zinc fin Hs.76526
er rotein Zf9 mRNA;
M97796 inhibitor of DNA bindin 2; dominant Hs.180919
ne ative helix-loo -helix
086782 26S roteasome-associated ad1 homolo Hs.178761
AA099391 ESTs Hs.77310
2 0 M19267 tro om osin 1 al ha Hs.77899
D29992 tissue factor athwa inhibitor 2 Hs.78045
L19314 hos ho lase kinase; beta Hs.195217
S78569 laminin; al ha 4 Hs.78672
028811 "Human c steine-rich fibroblast rowth
factor rece for CFR-1
2 5 L77886 rotein t rosine hos hatase; rece for Hs.79005
t e; K
C14407 neuronal tissue-enriched acidic roteinHs.79516
M60278 di htheria toxin rece for he arin-bindinHs.799
a idermal rowth
881509 s licin factor; ar inine/serine-rich Hs.184571
11
AA487558 ESTs Hs.8135
3 0 D86962 KIAA0207 ene roduct Hs.81875
AA478971 disabled Droso hila homolo 2 mito Hs.81988
en-res onsive
D50683 transformin rowth factor; beta rece Hs.82028
for II 70-80kD
056637 ca in rotein actin filament muscle Hs.184270
Z-line; al ha 1
M61199 Human cleava a si nal 1 rotein mRNA; Hs.82767
com lete cds
35 M28882 "Human MUC18 I co rotein mRNA, com
lete cds"
X15183 CDW52 anti en CAMPATH-1 anti en Hs.180532
S53911 CD34 Hs.85289
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
U20734 Human transcri tion factor'unB 'unB Hs.198951
ene; 5' re ion and
D28235 rosta landin-endo eroxide s nthase Hs.92309
2 rosta landin G/H
AA236324 ESTs; Weakl similar to !!!! ALU CLASSHs.92381
A WARNING ENTRY !!!!
AA148923 Homo sa iens mRNA for DEPP decidual Hs.93675
rotein induced b
AA174183 ESTs Hs.93872
AA456311 ESTs; Weakl similar to !!!! ALU CLASSHs.93961
A WARNING ENTRY !!!!
L08069 heat shock rotein; DNAJ-like 2 Hs.94
AA452000 ESTs Hs.94030
AA282140 ESTs Hs.9587
J02854 m osin re ulato Ii ht chain 2; smoothHs.9615
muscle isoform
AA442054 hos holi ase C; amma 1 formerl subt Hs.993
a 148
AB000450 vaccinia related kinase 2
AB002380 KIAA0382 rotein
AB003103 roteasome rosome; macro ain 26S subunit;
non-ATPase; 12
AB004884 tousled-like kinase 2
AF000573 homo entisate 1;2-diox enase homo
entisate oxidase
AF008937
AF009301 similar to S. cerevisiae SSM4
AF009368 cAMP res onsive element bindin rotein
3 luman
2 0 D00591 chromosome condensation 1
D00760 roteasome rosome; macro ain subunit;
al ha t e; 2
D11139 tissue inhibitor of metallo roteinase
1 a hyoid otentiatin
D14657 KIAA0101 ene roduct
D14878 D123 ene roduct
mannosyl (alpha-1;6-)-glycoprotein
D17716 beta-1;6-N-acet I- lucosamin Itransferase
D21090 RAD23 S. cerevisiae homolo B
D26135 diac I I cerol kinase; amma 90kD
D26528 DEAD/H As -Glu-Ala-As /His box of
a tide 7 RNA helicase;
D30742 calcium/calmodulin-de endent rotein
kinase IV
3 0 D31762 KIAA0057 ene roduct; TRAM-like rotein
D31765 KIAA0061 rotein
D31888 KIAA0071 rotein
D38128 rosta landin 12 rostac clin rece for
IP
D38500 ostmeiotic se re ation increased 2-like
4
3 5 D38551 RAD21 S. ombe homolo
D42087 KIAA0118 rotein
D49396 antioxidant rotein 1
D55640
D63391 latelet-activatin factor acet Ih drolase;
isoform Ib; amma
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Exemplar
Accession Com lete Title UniGeneID 11/29/99
D63477 KIAA0143 rotein
D63483 acet I LDL rece tor; SREC
D64015 TIA1 c otoxic ranule-associated RNA-bindin
rotein-like 1
D79990 KIAA0168 ene roduct
D79997 KIAA0175 ene roduct
D80010 KIAA0188 rotein
D84276 CD38 anti en 45
D86425 nido en 2
D86978 KIAA0225 rotein
D87012 Human lambda DNA for immuno lobin
Ii ht chain
D87075 solute carrier famil 23 nucleobase
traps orters ; member 1
D87432 solute carrier famil 7 cationic amino
acid traps orter; +
D87448 to oisomerase DNA II bindin rotein
D87845 latelet-activatin factor acet Ih drolase
2 40kD
HG1098-HT1098
HG2167-HT2237
HG2415-HT2511
HG2825-HT2949
HG2887-HT3031
2 0 HG4660-HT5073
HG4704-HT5146
HG884-HT884
HG919-HT919
J00212
2 5 J04029 keratin 10 a idermol is h erkeratosis;
keratosis almaris et
methylenetetrahydrofolate dehydrogenase
J04031 (NADP+ dependent);
methen Itetrah drofolate c cloh drolase;
form Itetrah drofolate
J04088 to oisomerase DNA II al ha 170kD
J04543 annexin A7
TEK tyrosine kinase; endothelial (venous
L06139 malformations; multiple
cutaneous and mucosal
3 0 L07540 re lication factor C activator 1 5
36.5kD
L08895 MADS box transcri tion enhancer factor
2; of a tide C
L11239 astrulation brain homeo box 1
L11353 neurofibromin 2 bilateral acoustic
neuroma
myeloid/lymphoid or mixed-lineage
L13773 leukemia (trithorax
Droso hila homolo ; translocated to;
2
3 5 L13800
L14922 re lication factor C activator 1 1
145kD
L15189 heat shock 70kD rotein 9B mortalin-2
L15388 G rotein-cou led rece for kinase 5
L16895 I s I oxidase
4 0 L27476 ~ ight junction protein 2 (zona occludens
t 2)
135
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
L27624 tissue factor athwa inhibitor 2
L32976 mito en-activated rotein kinase kinase
kinase 11
L33404 kallikrein 7 ch mot tic; stratum corneum
L35263 mito en-activated rotein kinase 14
L37347 solute carrier Tamil 11 roton-cou
led divalent metal ion
L40371 th roid hormone rece for interactor
4
L40391 Homo sa iens clone s153 mRNA fra ment
L41607 lucosamin I N-acet I transferase 2;
I-branchin enz me
L77566 DiGeor a s ndrome critical re ion
ene DGSI
M13928 aminolevulinate; delta-; deh dratase
M13928 aminolevulinate; delta-; deh dratase
M14016 uro or h rino en decarbox lase
M14219 decorin
M15796 roliferatin cell nuclear anti en
M21305 Human al ha satellite and satellite
3 'unction DNA se uence
M22092
M22898 tumor rotein 53 Li-Fraumeni s ndrome
M22995 RAP1A; member of RAS onto ene famil
M23379 RAS 21 rotein activator GTPase activatin
rotein 1
2 0 M24364 ma'or histocom atibilit tom )ex; class
I I; DQ beta 1
M24400 ch mot sino en B1
M25753 c clin B1
M27691 cAMP res onsive element bindin rotein
1
M28213 RAB2; member RAS onto ene famil
2 5 M29550 rotein hos hatase 3 former) 2B ; catal
is subunit; al ha
M29971 O-6-meth I uanine-DNA meth Itransferase
M30269 nido en enactin
M31158 rotein kinase; cAMP-de endent; re
ulato ; t a II; beta
M31166 entaxin-related ene; ra id) induced
b IL-1 beta
endothelial differentiation; sphingolipid
3 0 M31210 G-protein-coupled
rece tor; 1
M55420 E silon ; I E
M59979 rosta landin-endo eroxide s nthase
1 rosta landin G/H
M62810 transcri tion factor 6-like 1 mitochondria)
transcri tion factor
M63838 interferon; amma-inducible rotein
16
3 5 M64710 Human CNP ene for C-t a natriuretic
a tide
M68874
M74524 ubi uitin-con'u atin enz me E2A RAD6
homolo
M80254 a tid I rol I isomerase F c clo hilin
F
M81780 s hin om elfin hos hodiesterase 1;
acid I sosomal acid
40 M81780 s hin om elfin hos hodiesterase 1;
acid I sosomal acid
136
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
M81780 s hin om elfin hos hodiesterase 1;
acid I sosomal acid
Homo Sapiens acid sphingomyelinase
M81780 (SMPD1 ) gene; complete
cds; ORF's 1-3; com lete cds's
Homo Sapiens acid sphingomyelinase
M81780 (SMPD1 ) gene; complete
cds; ORF's 1-3; com lete cds's
M83822 cell division c cle 4-like
M86934 DNA se ment; numerous co ies; ex ressed
robes GS1 ene
M87338 re lication factor C activator 1 2
40kD
M96326 azurocidin 1 cationic antimicrobial
rotein 37
M96954 TIA1 c otoxic ranule-associated RNA-bindin
rotein-like 1
M98833 Friend leukemia virus inte ration
1
S66793 arrestin 3; retinal X-arrestin
S72370 ruvate carbox lase
S78569 laminin; al ha 4
S79873 I sosomal-associated membrane rotein
2
S83325 as artate beta-h drox lase
S83364
S83365
001212 olfacto marker rotein s mbol rovisional
001922 translocase of inner mitochondria)
membrane 8 east homolo A
002556 t-com lex-associated-testis-ex ressed
1-like
2 0 002680 rotein t rosine kinase 9
003272 fibrillin 2 con enital contractural
arachnodact I
004209 microfibrillar-associated rotein 1
005237 fetal Alzheimer anti en
007225 uriner is rece for P2Y; G- rotein
cou led; 2
2 5 007620 mito en-activated rotein kinase 10
009759 mito en-activated rotein kinase 9
009820 al ha thalassemia/mental retardation
s ndrome X-linked
011313 sterol carrier rotein 2
014518 centromere rotein A 17kD
30 014575 rotein hos hatase 1; re ulato inhibitor
subunit 8
015173 BCL2/adenovirus E1B 19kD-interactin
rotein 2
015932 dual s ecificit hos hatase 5
018291 CDC16 cell division c cle 16; S. cerevisiae;
homolo
018300 dama e-s ecific DNA bindin rotein
2 48kD
3 5 018383 nuclear res irato factor 1
020536 cas ase 6; a o tosis-related c steine
rotease
021551 branched chain aminotransferase 1;
c osolic
023028 euka otic translation initiation factor
2B; subunit 5 a silon;
023752 SRY sex-determinin re ion Y -box 11
4 0 025435 t ranscri tional re ressor
025997 stanniocalcin
137
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
028251 zinc fin er rotein 169
028831
030245
032315 s ntaxin 3A
032439 re ulator of G- rotein si nallin 7
032849 N-m c and STAT interactor
035139 necdin mouse homolo
036764 euka otic translation initiation factor
3; subunit 2 beta; 36kD
039400 chromosome 11 o en readin frame 4
039657 mito en-activated rotein kinase kinase
6
041344 roline ar inine-rich end leucine-rich
re eat rotein
041766 a disinte rin and metallo roteinase
domain 9 meltrin amma
041813 homeo box A9
041815 nucleo orin 98kD
043286 seleno hos hate s nthetase 2
044378 MAD mothers a ainst deca enta 1e ic;
Droso hila homolo 4
044754 small nuclear RNA activatin com lex;
of a tide 1; 43kD
047011 fibroblast rowth factor 8 andro en-induced
047011 fibroblast rowth factor 8 andro en-induced
2 0 047011 fibroblast rowth factor 8 andro en-induced
047011 fibroblast rowth factor 8 andro en-induced
047077 rotein kinase; DNA-activated; catal
is of a tide
048251 rotein kinase C bindin rotein 1
050535 Human BRCA2 re ion; mRNA se uence CG006
2 5 056833 von Hi el-Lindau bindin rotein 1
058091 cullin 4B
058837 c clic nucleotide ated channel beta
1
059289 cadherin 13; H-cadherin heart
059863 TRAF famil member-associated NFKB activator
3 0 067122 ubi uitin-like 1 sentrin
067319 cas ase 7; a o tosis-related c steine
rotease
068019 MAD mothers a ainst deca enta 1e ic;
Droso hila homolo 3
a disintegrin and metalloproteinase
069611 domain 17 (tumor necrosis
factor; al ha; convertin enz me
070322 ka o herin im ortin beta 2
3 5 073524 ATP/GTP-bindin rotein
079267 rotein hos hatase 4; re ulato subunit
1
079291 Human clone 23721 mRNA se uence
082671 Homo sa iens clone LM1955 H105e3 ene;
artial cds
082671 zinc fin er rotein 185 LIM domain
4 0 084573 rocolla en-I sine; 2-oxo lutarate 5-diox
enase I sine
090914 carbox a tidase D
091316 c osolic ac I coenz me A thioester
h drolase
091932 ada tor-related rotein com lex 3; si
ma 1 subunit
138
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
096131 Homo sa iens HPV16 E1 rotein bindin
rotein mRNA;
097018 echinoderm microtubule-associated rotein-like
097188 IGF-II mRNA-bindin rotein 3
V00503 colla en; t a I; al ha 2
X04327 2;3-bis hos ho I cerate mutase
X06389 s na to h sin
X07496 a oli o rotein A-I
X07820 matrix metallo roteinase 10 stromel
sin 2
X14787 thrombos ondin 1
X15525 acid hos hatase 2; I sosomal
methylene tetrahydrofolate dehydrogenase
X16396 (NAD+ dependent);
methen Itetrah drofolate c cloh drolase
X16609 ank rin 1; a hroc is
X53586 inte rin; al ha 6
X53586 inte rin; al ha 6
X53793 multifunctional of a tide similar to
SAICAR s nthetase and AIR
X54936 lacental rowth factor; vascular endothelial
rowth factor-related
X55740 5' nucleotidase CD73
X57025 insulin-like rowth factor 1 somatomedin
C
X60673 aden late kinase 3
2 0 X60673 aden late kinase 3
X60708 di a tid I a tidase IV CD26; adenosine
deaminase com lexin
X62048 wee1+ S. ombe homolo
X63097 Rhesus blood rou ; D anti en
X63563 0l merase RNA II DNA directed of a
tide B 140kD
2 5 X64037 eneral transcri tion factor IIF; of
a tide 1 74kD subunit
X69636 hect domain and RLD 2
X69878 fms-related t rosine kinase 4
X70649 DEAD/H As -Glu-Ala-As /His box of a
tide 1
X72841 retinoblastoma-bindin rotein 7
3 0 X74987 ATP-bindin cassette; sub-famil E OABP
; member 1
X83107 BMX non-rece for t rosine kinase
X84194 ac I hos hatase 1; a hroc a common
t a
X85753 c clin-de endent kinase 8
X87870 he atoc a nuclear factor 4; al ha
3 5 X89066 transient rece for otential channel
1
X89398 uracil-DNA I cos lase
X89398 uracil-DNA I cos lase
X89399 RAS 21 rotein activator GTPase activatin
rotein 3
X89426 endothelial cell-s ecific molecule
1
4 0 X91247 t hioredoxin reductase 1
X91648 H.sa iens mRNA for ur al ha extended
3'untranslated re ion
139
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
X92098 coated vesicle membrane rotein
X92110 H.sa lens mRNA for he VIII rotein
X94703 RAB28; member RAS onco ene famil
X96506 DR1-associated rotein 1 ne ative cofactor
2 al ha
X97230 killer cell immuno lobulin-like rece
tor; three domains; Ion
X98263 M- hase hos ho rotein 6
X98296 ubi uitin s ecific rotease 9; X chromosome
Droso hila fat
X99584 SMT3 su ressor of mif two 3; east homolo
1
Y00264 am loid beta A4 recursor rotein rotease
nexin-II; Alzheimer
Y07566 Ric Droso hila -like; ex ressed in
man tissues
Y07759 m osin VA heav of a tide 12; m oxin
Y07827 but ro hilin; subfamil 3; member A1
Y07867 Pirin
Y09443 alk I I cerone hos hate s nthase
Y09858 H.sa lens mRNA for unknown rotein
Y12394 ka o herin al ha 3 im ortin al ha 4
211559 iron-res onsive element bindin rotein
1
211695 mito en-activated rotein kinase 1
215005 centromere rotein E 312kD
2 0 246261 H3 histone famil ; member A
AA011243 0l rC -bindin rotein 2
ESTs; Weakly similar to type-1 protein
AA018418 phosphatase skeletal
muscle I co en tar etin subunit H.sa
iens
AA018758 ESTs
AA018804 Homo sa iens clone 23675 mRNA se uence
2 5 AA031993 SUMO-1 activatin enz me subunit 2
AA044217 ESTs; Weakl similar to colla en al
ha 2 I chain R.norve icus
SWI/SNF related; matrix associated;
AA046548 actin dependent regulator of
chromatin; subfamil e; member 1
AA057447 ESTs; Moderatel similar to alternativel
s liced roduct usin
AA058376 S'o ren s ndrome anti en A2 60kD; ribonucleo
rotein
3 0 AA083572 v-ral simian leukemia viral onco ene
homolo A ras related
AA085696
AA088744 ESTs
AA089688 EST
AA091284 ESTs; Hi hl similar to HSPC030 H.sa
iens
3 5 AA092700 ESTs
AA092968 ESTs
AA094800 euka otic translation initiation factor
3; subunit 7 zeta; 66/67kD
AA100219 ESTs
AA114885 ESTs
140
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA129547 met roto-onco ene he atoc a rowth factor
rece for
AA133016 ESTs
AA149507 homolo of mouse uakin QKI KH domain
RNA bindin rotein
AA151005 s erm surface rotein
AA187101
AA195179 euka otic translation initiation factor
4A; isoform 2
AA203138 low densit Ii o rotein rece for familial
h ercholesterolemia
AA203645 Ar /Abl-interactin rotein Ar BP2
AA206236
AA227621 ESTs; Weakl similar to weak similarit
to colla ens C.ele ans
AA248283 ESTs; Weakl similar to rostate-s ecific
traps lutaminase
AA249611 SH3-bindin domain lutamic acid-rich
rotein
AA282640 ubi uitination factor E4B homolo ous
to east UFD2
AA287199 KIAA0081 rotein
AA313990 DKFZP564M112 rotein
AA314256 ESTs; Hi hl similar to CGI-94 rotein
H.sa iens
AA314389 ADP-ribos lation factor-like 5
AA324364 ESTs
AA329211 NS1-associated rotein 1
2 0 AA399187 DKFZP434A043 rotein
AA421079 ESTs; Weakl similar to Sox-like transcri
tional factor H.sa iens
AA422029 ESTs
AA425230 Ras-GTPase-activatin rotein SH3-domain-bindin
rotein
AA447052 KIAA0251 rotein
AA452000 Homo sa iens mRNA; cDNA DKFZ 586E1624
from clone
AA456687 ESTs
AA487015 Homo sa iens mRNA; cDNA DKFZ 586L0120
from clone
AB002326 Human mRNA for KIAA0328 ene; artial
cds
-BioB-3
3 0 C01527 ESTs
C01714 serum-inducible kinase
C01811 Homo sa iens clone 24921 mRNA se uence
C02352 ESTs; Hi hl similar to CGI-121 rotein
H.sa iens
C02375 ESTs
3 5 C14448 EST
D16611 co ro or h rino en oxidase co ro or
h ria; hardero or h ria
D25216 KIAA0014 ene roduct
D31352 ESTs
D58024 ESTs; Weakl similar to KIAA0768 rotein
H.sa iens
4 0 D80897 KIAA1036 rotein
~ D82614 ~ ESTs
141
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
D87845 latelet-activatin factor acet Ih drolase
2 40kD
D89377 msh Droso hila homeo box homolo 2
H06583 cAMP res onsive element bindin rotein-like
2
H40732 ESTs
H46617 ESTs
H56731 ESTs
H75570 ESTs
H78886 ESTs
H81241 Kru el-like factor 8
L36531 inte rin; al ha 8
M63154 astric intrinsic factor vitamin B s
nthesis
M63180 threon I-tRNA s nthetase
M91504 ESTs
N56191 rotocadherin 68
N78483 ESTs; Weakl similar to F20D12.3 ene
roduct C.ele ans
N79268 zinc fin er rotein 198
814652 Homo sa iens PAC clone DJ0872F07 from
7 31
820459 ESTs
822303 ESTs; Weakl similar to utative 150
H.sa iens
2 0 833779 ESTs; Weakl similar to 40 H.sa iens
836553 ESTs; Weakl similar to KIAA0681 rotein
H.sa lens
864534 ESTs
866475 ESTs
870621 KIAA0896 rotein
2 5 879356 KIAA0544 rotein
884933 ESTs
AA007160 Homo sa iens mRNA; cDNA DKFZ 564D016
from clone
AA007234 ESTs
AA018409 ESTs
3 0 AA025351 ESTs
AA027168 KIAA0955 rotein
AA027317 ESTs
ESTs; Weakly similar to PUTATIVE PRE-MRNA
AA029423 SPLICING
FACTOR RNA HELICASE H.sa iens
AA031357 ESTs; Weakl similar to N-WASP H.sa
iens
3 5 AA045136 ESTs
AA053400 ESTs
AA055829 ESTs; Weakl similar to !!!! ALU SUBFAMILY
J WARNING
AA065217 ESTs
AA116054 ESTs; Weakl similar to KIAA0638 rotein
H.sa iens
4 0 AA126311 ESTs
AA129390 ESTs
~ AA130273 ESTs; Weakly similar to hypothetical
protein; similar to
142
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA142919 ESTs
AA150205 Kru el-like factor 7 ubi uitous
AA176867 ESTs
AA180321 Homo sa iens clone S164 mRNA; 3' end
of cds
AA180487 transformin ; acidic coiled-coil containin
rotein 1
AA187634 euka otic translation initiation factor
3; subunit 1 al ha; 35kD
AA195399 ESTs
AA234717 ESTs
AA234743 ESTs
AA234957 m otubularin related rotein 1
AA235604 Homo sa iens clone 25007 mRNA se uence
AA236559 ESTs; Weakl similar to !!!! ALU SUBFAMILY
SQ WARNING
AA242868 ESTs; Weakl similar to house-kee in
rotein M.musculus
AA251776 'un D roto-onco ene
AA251909 buddin uninhibited b benzimidazoles
1 east homolo ; beta
diptheria toxin resistance protein
AA252672 required for diphthamide
bios nthesis Saccharom ces -like 2
AA256157 ESTs
AA256680 Homo sa iens mRNA; cDNA DKFZ 564H1916
from clone
AA258873 ESTs
2 0 AA262727 KIAA1033 rotein
AA281451 DKFZP564A043 rotein
AA281545 nuclear rece for co-re ressor 1
AA282069 KIAA0603 ene roduct
AA283044 ESTs
2 5 AA283930 ESTs
AA284755 CDW52 anti en CAM PATH-1 anti en
AA291268 DKFZP586L0724 rotein
AA291927 ESTs
AA343514 ESTs
3 0 AA398109 ESTs
AA405737 ESTs
AA406610 ESTs
AA411465 ESTs; Moderatel similar to HMG-box
transcri tion factor
AA416886 Homo sa iens mRNA; cDNA DKFZ 564C1563
from clone
3 5 AA424013 Homo sa iens clone 23767 and 23782
mRNA se uences
AA424148 DKFZP4341116 rotein
AA424558 hosducin-like
AA424961 similar to S. cerevisiae SSM4
AA425367 ESTs
4 0 AA425921 ESTs
AA426220 KIAA0523 rotein
143
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA427735 ESTs
AA430673 ESTs
AA432248 ESTs
AA435896 ESTs
AA436705 KIAA0766 ene roduct
AA446561 KIAA0470 ene roduct
AA448238 KIAA0915 rotein
AA448688 ESTs; Weakl similar to KIAA0638 rotein
H.sa iens
AA449756 ESTs; Weakl similar to !!!! ALU SUBFAMILY
J WARNING
1 0 AA450303 ESTs
AA452411 ESTs; Hi hl similar to mediator H.sa
iens
AA454566 hemo lobin; amma G
AA454667 ESTs
AA456437 ESTs
1 5 AA456646 ESTs
AA456826 ESTs
AA456981 ESTs
AA458959 ESTs
AA459950 ESTs
2 0 AA460449 ESTs; Hi hl similar to hos hoserine
aminotransferase
AA463910 ESTs
AA464603 ESTs
AA464606 MRS1 rotein
AA465093 TIA1 c otoxic ranule-associated RNA-bindin
rotein
2 5 AA465692 KIAA0648 rotein
AA476473 tri 1e functional domain PTPRF interactin
AA478109 ESTs
AA478474 ESTs
AA480889 ESTs
3 0 AA485223 ESTs
AA485254 ESTs
AA486183 ESTs; Weakl similar to similar to ox
sterol-bindin roteins
AA496936 ESTs
AA598589 ESTs
3 5 AA598831 ESTs
AA600150 ESTs
AA608545 RAD51 S. cerevisiae homolo E coli RecA
homolo
AA609210 ESTs
AA610108 ESTs; Hi hl similar to CGI-124 rotein
H.sa iens
4 0 AA620582 ESTs; Weakl similar to KIAA0869 rotein
H.sa lens
AA621239 ESTs; Hi hl similar to ALG-2 interactin
rotein AIP1
AA621714 ESTs
144
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Exemplar
Accession Com lete Title UniGeneID 11/29/99
AA621718 ESTs; Moderatel similar to CGI-74
rotein H.sa iens
D19673 ESTs
D25755 ESTs
D51095 DKFZP586E1621 rotein
D60272 ESTs; Weakl similar to macro ha a
lectin 2 H.sa iens
T08879 cathe sin F
UDP-N-acetyl-alpha-D-galactosamine:polypeptide
T34527 N-acet I alactosamin Itransferase
1 GaINAc-T1
T40327 lun resistance-related rotein
T62771 nucleo hosmin/nucleo lasmin; 3
T63174 Homo sa iens mRNA; cDNA DKFZ 58610324
from clone
T83444 KIAA0887 rotein
T93641 ESTs
048263 re ronocice tin
049065 interleukin 1 rece tor-like 2
079300 Human clone 23629 mRNA se uence
088573 NBR2
093867 0l merase RNA III DNA directed 62kD
W01094 ESTs
W01568 ESTs
cartilage oligomeric matrix protein
W26853 (pseudoachondroplasia;
a i h seal d s lasia 1; multi 1e
W27179 BCL2/adenovirus E1 B 19kD-interactin
rotein 3-like
W27965 EST
W36280 NS1-associated rotein 1
W47063 ESTs
2 5 W79060 isocitrate deh dro enase 2 NADP+ ;
mitochondrial
W88550 KIAA1058 rotein
X60486 H4 histone famil ; member G
X78931 zinc fin er rotein 272
214077 YY1 transcri tion factor
3 0 AA002147 EST
AA004711 ESTs
AA010383 EST
AA015761 ESTs
AA018772 ESTs
3 5 AA021473 EST
AA024835 otassium volta e- ated channel; dela
ed-rectifier; subfamil S;
AA025858 Homo sa iens mRNA; cDNA DKFZ 58681024
from clone
AA027229 ESTs; Weakl similar to F45E12.5 C.ele
ans
AA029428 ESTs
4 0 AA035143 ESTs
~ AA035237 butyrate response factor 2 (EGF-response
I factor 2)
145
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA039347 EST
AA040740 ESTs
AA041551 ESTs
AA045513 ESTs
AA045745 ESTs
AA055348 ESTs
AA056582 KIAA0372 ene roduct
AA056697 ESTs
AA056746 EST
1 0 AA057678 ESTs
AA058681 ESTs
AA058686 ESTs
AA062840
AA064859
1 5 AA065069
AA069923
AA070799 zinc fin er rotein 6 CMPX1
AA070815
AA075374
2 0 AA076382
AA078787 ESTs
AA078986
AA079393
AA079487
2 5 AA083207 EST
AA083256
AA084415
AA085274
AA088678 ESTs
stress-associated endoplasmic reticulum
3 0 AA100925 protein 1; ribosome
associated membrane rotein 4
AA101255 Homo sa iens mRNA for H-2K bindin factor-2;
com lete cds
AA126474 stanniocalcin 2
AA127017 ESTs
ESTs; Weakly similar to PROTEIN PHOSPHATASE
AA129968 PP2A; 130
KD REGULATORY SUBUNIT H.sa iens
3 5 AA130240 ESTs
AA131866 ESTs; Weakl similar to DY3.6 C.ele
ans
AA132039 ESTs
AA132983 DKFZP586G1517 rotein
AA133250 ESTs
4 0 AA133583 hi h-mobilit rou nonhistone chromosomal
rotein isoform I-C
AA135941 ESTs
' AA148650 - _ _
-~
146
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Exemplar
Accession Com lete Title UniGeneID 11/29/99
AA151110 ESTs
AA155754 EST
ESTs; Moderately similar to hedgehog-interacting
AA156125 protein
M.musculus
AA156289 ESTs
AA156997 ESTs
AA157291 EST
AA157293 ESTs
AA164293 ESTs
ESTs; Weakly similar to weak similarity
AA164676 to S. cerevisiae
intracellular rotein traps ort rotein
US 1 C.ele ans
AA167375 KIAA0530 rotein
AA167550 Homo sa iens mRNA; cDNA DKFZ 564M113
from clone
AA176589 EST
AA180448 EST
AA187144 endothelin 1
1 5 AA189170 ESTs
AA192757 ESTs
AA205650 ESTs
AA233342 ESTs; Weakl similar to WD40 rotein
Ciao 1 H.sa iens
AA233472 ESTs
2 0 AA234110 ESTs
D80981 ESTs
F01660 ESTs
F02206 EST; Hi hl similar to ether-a- o-
o-related rotein H.sa lens
F02208 ESTs
2 5 F02544 ESTs
F03918 ESTs
F04258 ro hos hatase inor anic
F04600 ESTs
F08998 ESTs
3 0 F09605 ESTs
F11115 ESTs
H06371 Homo sa iens clone 24993 mRNA se uence
H10995 Homo sa iens mRNA full len th insert
cDNA clone EUROIMAGE
H11938 ESTs; Hi hl similar to histone acet
Itransferase H.sa lens
3 5 H16568 ESTs
H16772 ESTs
H18951 ESTs; Moderatel similar to dJ1163J1.1
H.sa iens
H20859 ESTs
H23747 ESTs
4 0 H38087 ESTs; Weakl similar to NG22 H.sa iens
H40331 ESTs
H40567 ~ ESTs _ _ ~ _
147
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
H46966 ESTs
H56640 ESTs
H57154 ESTs; Weakl similar to or anic anion
traps orter 1 H.sa iens
H96712 ESTs
N20814 ESTs
N25249 s pa tosomal-associated rotein; 23kD
N27100 keratin 5 a idermol sis bullosa sim
lex;
N39616 RNA uanine-7- meth Itransferase
N48982 ESTs
1 0 N51957 ESTs
N52271 LIM rotein similar to rat rotein kinase
C-bindin eni ma
N59435 ESTs; Hi hl similar to CGI-112 rotein
H.sa lens
N64139 ESTs; Weakl similar to lar a tumor
su ressor 1 H.sa iens
N66981 ESTs
1 5 N68640 ESTs
N69352 DEAD/H As -Glu-Ala-As /His box of a
tide 15
N95226 KIAA0758 rotein
800138 ESTs
807998 ESTs; Weakl similar to !!!! ALU SUBFAMILY
J WARNING
2 0 808929 ubi uitin-con'u atin enz me E2G 2 homolo
ous to east UBC7
810307 ESTs
833354 ESTs
836083 ESTs
837938 KIAA0440 rotein
2 5 839330
840816 cullin 4A
843162 ESTs
845698 ESTs; Weakl similar to cAMP inducible
2 rotein M.musculus
854554 ESTs
3 0 868425 ESTs
868568 ATX1 antioxidant rotein 1; east homolo
1
868763 ESTs
870467 ESTs
873565 Homo sa iens mRNA; cDNA DKFZ 564M113
from clone
3 5 873640 ESTs
878376 EST
892453 EST
T03865 ESTs
T03872 ESTs
4 0 T10072 ESTs
T10080 ESTs
T10132 KIAA0478 gene product
148
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Exemplar
Accession Com lete Title UniGeneID 11/29/99
T15343 ESTs
T23457 ESTs
T23555 ESTs
T23670 ESTs
T23948 ESTs
T33464 ESTs
T34413 ESTs
T34611 ESTs
T40920 ESTs
T55182 ESTs; Hi hl similar to IGF-II mRNA-bindin
rotein 2 H.sa iens
T77453 ESTs
T84039 ESTs
T86458 ESTs
T87693 EST
1 5 T89350 ESTs
T90945 ESTs
T90987 ESTs
T91863 ESTs
T91881 KIAA0563 ene roduct
2 0 T93783 ESTs
T96687 ESTs
T96944 Homo sa iens mRNA; cDNA DKFZ 434H132
from clone
T97307 ESTs; Moderatel similar to !!!! ALU
SUBFAMILY J WARNING
T97764 ESTs
2 5 W48817 ESTs
W58343 DKFZP586B2420 rotein
W59949 ESTs; Moderatel similar to GTP-BINDING
PROTEIN TC10
W74644 ESTs
W74761 ESTs; Hi hl similar to ubi uitin-con'u
atin enz me HBUCE1
3 0 W74802 ESTs
W81205 ESTs
W81237 ESTs
W90146 ESTs
W92798 ESTs
3 5 238412 EST
238709 inositol 1;4;5-tri hos hate rece tor;
t a 2
238904 ESTs; Weakl similar to KIAA0970 rotein
H.sa lens
239103 core-bindin factor; runt domain; al
ha subunit 2; translocated to;
239930 calreticulin
4 0 239939 ESTs
2 40012 NCK-associated rotein 1
'2 40377 I ESTs
149
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Exemplar
Accession Com lete Title UniGeneID 11/29/99
240820 ESTs
241680 Homo sa iens mRNA; cDNA DKFZ 566P013
from clone
-BioB-3
AA005112 LIM domain onl 7
AA005432 DKFZP547E2110 rotein
AA010163 a stream re ulato element bindin rotein
1
AA026356 ESTs
AA026901 ESTs
AA036867 ESTs; Weakl similar to coded for b
C. ele ans cDNA k30b3.5
AA044644 I m hoc e-s ecific rotein 1
AA046426 Cdc42 effector rotein 3
AA054515 ESTs; Weakl similar to rostate-s ecific
traps lutaminase
AA084162
AA085749 ATP bindin rotein associated with
cell differentiation
AA098874 DKFZP4341116 rotein
AA 101056
AA102746 ESTs
AA114250 KIAA0512 ene roduct
AA126561 stanniocalcin
2 0 AA128980 ESTs
AA129757 ESTs; Weakl similar to 60S RIBOSOMAL
PROTEIN L22
AA129921 S-adenos Ihomoc steine h drolase-like
1
AA133331 KIAA0741 ene roduct
AA135958 ESTs
2 5 AA136524 euka otic translation elon ation factor
1 al ha 1
AA147044 ESTs; Weakl similar to !!!! ALU CLASS
C WARNING ENTRY !!!!
AA148885 minichromosome maintenance deficient
S. cerevisiae 4
AA150043 ESTs
AA151621 ESTs
3 0 AA155743 ferritin; Ii ht of a tide
AA156335 ESTs
AA156336 nuclear rece for co-re ressor 1
AA159181 ESTs; Weakl similar to L a8 S.cerevisiae
AA159825 ESTs; Weakl similar to ORF YNL227c
S.cerevisiae
3 5 AA234185 ESTs
AA234929 ESTs
AA234935 ESTs
AA236359 ESTs
AA236466 ESTs
4 0 AA236535 Human clone 23654 mRNA se uence
AA236935 Human normal keratinoc a mRNA
~ AA236942 ESTs
~
150
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Exemplar
Accession Com lete Title UniGeneID 11/29/99
AA237018 ESTs
AA237025 ESTs
AA242751 KIAA0903 rotein
AA242760
AA242763 CDC14 cell division c cle 14; S. cerevisiae
homolo B
AA242809 ESTs; Weakl similar to !!!! ALU SUBFAMILY
J WARNING
AA243133 serine/threonine kinase 15
AA243495 lectin; mannose-bindin ; 1
AA243706 ESTs
1 0 AA250848 ESTs
AA250868 ESTs
AA251152 ESTs
AA251544 ESTs
AA251792 fatt -acid-Coenz me A Ii ase; Ion
-chain 4
AA252063 BH- rotocadherin brain-heart
AA252144 ESTs
AA252524
AA253461 ESTs
AA255522 ESTs; Weakl similar to INHIBITOR OF
APOPTOSIS PROTEIN 1
2 0 AA256468 ESTs
AA256528 ESTs
AA257976 ESTs
AA258296 KIAA0579 rotein
AA258409 m elfin rotein zero-like 1
2 5 AA258421 h othetical rotein
AA262077 aldeh de deh dro enase 5 famil ; member
A1
AA278650 ESTs; Weakl similar to similar to
the beta transducin famil
AA278766 ESTs
AA279667 natural killer-tumor reco nition se
uence
3 0 AA280791 euka otic translation initiation factor
5
AA280819 MADS box transcri tion enhancer factor
2; of a tide C
AA280828 Homo sa iens mRNA; cDNA DKFZ 586M141
from clone
AA282195 ESTs; Weakl similar to Unknown H.sa
iens
AA283127 Homo sa iens clone LM1955 H105e3 ene;
artial cds
3 5 AA284694 nucleo orin-like rotein 1
AA291137 ESTs
AA291708 ESTs; Weakl similar to !!!! ALU SUBFAMILY
SO WARNING
AA293495 chromosome 8 o en readin frame 1
AA347193 ESTs
4 0 AA398474 Homo sa iens mRNA; cDNA DKFZ 586H051
from clone
~ AA398512 STs
~
151
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA400277 ESTs
AA400896 ESTs
AA404494 CTP s nthase
AA410345 ESTs; Weakl similar to 'unctional adhesion
molecule H.sa iens
AA416733 ESTs; Weakl similar to !!!! ALU SUBFAMILY
SC WARNING
AA425154 ESTs
AA426573 ESTs; Moderatel similar to endomucin
M.musculus
AA431418 N-acet I lucosaminidase; al ha- Sanfili
o disease IIIB
Human DNA sequence from clone 44A20
on chromosome
6q23.1-24.3. Contains a gene for a
A436182 novel protein similar to
MTHFD1 (methylenetetrahydrofolate dehydrogenase
(NADP+
de endent ; methen Itetrah drofolate
c cloh drolase;
1 0 AA437099 ESTs
AA446585 ESTs
AA446887 ESTs
AA447224 ESTs; Weakl similar to cDNA EST CEESW54F
comes from this
AA447709 ESTs; Moderatel similar to utative
transcri tion factor CA150
AA453624 deox nucleotid Itransferase; terminal
AA455044 ESTs
AA456045 ESTs
AA460454 ESTs; Weakl similar to KIAA0512 rotein
H.sa iens
AA476494 ESTs; Weakl similar to KIAA0512 rotein
H.sa iens
2 0 AA476738 leucine rich re eat in FLII interactin
rotein 1
AA481422 Homo sa iens mRNA for H-2K bindin factor-2;
tom lete cds
AA482269 inte ral membrane rotein 1
AA482595 ESTs; Weakl similar to F25B5.3 C.ele
ans
AA485084 ESTs
2 5 AA485431 ESTs
AA489057 stromal anti en 2
AA489638 DKFZP564M2423 rotein
AA491000 Homo sa iens mRNA; cDNA DKFZ 586N1720
from clone
AA491250 ESTs
3 0 AA505133 solute carrier famil 2 facilitated
lucose traps orter ; member 3
AA598447 ex ortin; tRNA nuclear ex ort rece
for for tRNAs
AA599243 eneral transcri tion factor IIIA
AA599574 i ase; endothelial
l
AA600153 DEK onto ene DNA bindin
3 5 AA609309 ESTs; Weakl similar to !!!! ALU SUBFAMILY
J WARNING
AA609710 Human chromosome 3 21.1 ene se uence
AA610068 PIBF1 ene roduct
AA621399 ESTs
' AA621752 6S proteasome-associated padl homolog
~2
152
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Exemplar
Accession Com lete Title UniGeneID 11/29/99
C21523 ESTs
D12160 ESTs; Moderatel similar to !!!! ALU
SUBFAMILY J WARNING
D19708 ESTs
D25801 ESTs; Hi hl similar to KIAA0445 rotein
H.sa lens
a disintegrin-like and metalloprotease
D45652 (reprolysin type) with
thrombos ondin t a 1 motif; 4; a recap
1
D60208 ESTs
D80504 zinc fin er rotein 198
F03010 m eloid/I m hold or mixed-lines a
leukemia 2
F04247 ESTs
F10966 Homo sa iens mRNA; cDNA DKFZ 434M196
from clone
F13700 ribonuclease P; 40kD subunit
H05063 ESTs; Weakl similar to / rediction
H16758 a hro oietin rece for
H17315 EST
H22556 utative translation initiation factor
H22566 ESTs
H48459 KIAA0186 ene roduct
H53073
H56559 KIAA0601 rotein
2 0 H57957 EST
H64938 ESTs
H64973 ESTs
H69535 ESTs
H73110 ESTs
2 5 H81783 ESTs
H86259 Homo sa iens chromosome 19; cosmid
832611
H88353 ESTs; Weakl similar to line-1 rotein
ORF2 H.sa iens
H88639 YY1-associated factor 2
H88675 nuclear rece for co-re ressor 1
3 0 H93708 s erm s ecific anti en 2
N22107 ESTs
N24046 ESTs
N27028 ESTs
N30205 ESTs
3 5 N30621 ESTs
N33258 nuclear rece for co-re ressor 1
N33390 EST
N40180 EST
EST; Highly similar to similar to
N45198 Cdc14B1 phosphatase
H.sa iens
4 0 N45979 SH3 domain rotein 1 B
N48325 EST
153
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
N48913 ESTs
N49394 KIAA0716 ene roduct
N50656 ESTs; Hi hl similar to mosaic rotein
LR11 H.sa lens
N50721 si nal se uence rece tor; amma translocon-associated
rotein
N53143 Homo sa iens clone 25218 mRNA se uence
N53359 ESTs; Weakl similar to beta-TrCP rotein
E3RS-Ika aB
N55326 ESTs
N55493
N57493 EST
N62955 ESTs; Weakl similar to KIAA0396 H.sa
iens
N63520 EST
N63604 ESTs
N64166 frizzled Droso hila homolo 7
N64168 ESTs
N64191 ESTs
N66845 ESTs; Weakl similar to !!!! ALU CLASS
B WARNING ENTRY !!!!
N67135 ESTs
N67295 ESTs
N68399 H2B histone famil ; member N
2 0 N68963 ESTs
N69331 a tid I rol I isomerase C c clo hilin
C
N70777 ESTs
N71364 ESTs
N71545 ESTs
2 5 N71571 ESTs
N74456 EST
N75594 ESTs
N79035 ESTs
N80279 h othetical rotein
3 0 N91797 ESTs
N92454 ka o herin im ortin beta 1
N94581 actin; beta
N94746 ESTs
N98238 ESTs
3 5 802384 ESTs
ESTs; Weakly similar to !!!! ALU SUBFAMILY
816833 J WARNING
ENTRY !!!! H.sa iens
841828 m osin VA heav of a tide 12; m oxin
843203 EST
846395 DKFZP566A0946 rotein
4 0 858863 ESTs
878248 ESTs; Weakl similar to KIAA0970 rotein
H.sa iens
L T11483 _ ESTs - _
~
154
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Exemplar
Accession Com lete Title UniGeneID 11/29/99
T16896 ESTs
T23820 c clin T2
T30222 ESTs; Moderatel similar to tetrac
cline trans orter-like rotein
W15275 Homo sa iens mRNA; cDNA DKFZ 586E1624
from clone
W38194
W42414 MAD mothers a ainst deca enta 1e ic;
Droso hila homolo 3
W46577 endothelial cell-s ecific molecule
1
W49632 Human clone 23908 mRNA se uence
W57613 ESTs
W57759 EST
W61118 ESTs
W65344 ESTs; Moderatel similar to h othetical
rotein H.sa lens
W69216 ESTs
W69379 Homo sa iens mRNA; cDNA DKFZ 586D0923
from clone
W86728 ESTs
238499 MKP-1 like rotein t rosine hos hatase
238630 bladder cancer related rotein 10kD
239494 ESTs
239623 ESTs
2 0 240071 BMX non-rece for t rosine kinase
240174 ESTs
240182 EST
240904 EST
AA166965 ESTs
2 5 AA167500 EST
AA169599 ESTs
AA171724 ESTs; Weakl similar to ORF YNL059c
S.cerevisiae
AA171739 ESTs
ESTs; Weakly similar to MITOCHONDRIAL
AA177105 CARNITINE/ACYLCARNITINE CARRIER PROTEIN
H.sa lens
3 0 AA182626 ESTs
AA186324 cell c cle ro ression 8 rotein
AA192099 zinc fin er rotein 148 HZ-52
AA192173 ESTs
AA192415 ESTs
3 5 AA192553 ESTs; Hi hl similar to RGC-32 R.norve
icus
AA194851 ESTs
AA195520 ESTs
AA196300 ESTs; Weakl similar to alternativel
s liced roduct usin exon
AA196517 rotease; serine; 15
4 0 AA196549 ESTs
AA196721
155
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA196729 ESTs
AA196979 ESTs; Weakl similar to rotease H.sa
lens
AA206828
AA207123 immuno lobulin su ertamil ; member
3
AA214539 TIA1 c otoxic ranule-associated RNA-bindin
rotein
AA226914 nuclear rece for subfamil 2; rou C;
member 1
AA227260 Zic famil member 3 odd- aired Droso
hila homolo ; heterotax
AA227469 EST
ESTs; Highly similar to multifunctional
AA233122 calcium/calmodulin-de endent rotein
kinase II delta2 isoform
Machado-Joseph disease (spinocerebellar
AA233334 ataxia 3;
olivo ontocerebellar ataxia 3; autosomal
dominant; ataxin 3
AA233347 zinc fin er rotein 216
AA233519 ESTs; Weakl similar to evectin-1 R.norve
icus
AA233714 A 12 auto ha 12; S. cerevisiae -like
AA233796 euka otic translation initiation factor
4E
1 5 AA235050 ESTs
AA235704 ESTs; Weakl similar to Wiscott-Aldrich
S ndrome rotein
AA236031 ESTs
AA236352 ESTs
AA236390 ESTs
2 0 AA236453 ESTs
AA243370 EST
AA250947 ESTs
AA251083 ESTs
AA251113 ESTs
2 5 AA251973 ESTs
AA252023 ESTs; Weakl similar to HRIHFB2157 H.sa
iens
AA252414 ESTs
AA252650 mito en-activated rotein kinase kinase
7
AA255523 ESTs
3 0 AA258128 ESTs
AA262105 Homo sa iens mRNA; cDNA DKFZ 564L1916
from clone
AA262107 ESTs
AA262235 ESTs
AA278298 M- hase hos ho rotein 1
3 5 AA278529 serine/threonine kinase 18
AA278721 ESTs
AA280036 euka otic translation initiation factor
4A; isoform 2
AA280648 ESTs; Weakl similar to rab-related
~ GTP-bindin rotein
AA280738 ESTs
4 0 280794 ~ ESTs
156
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA280837 ESTs
AA280886 ESTs
AA280934 ESTs
AA281535 KIAA0879 rotein
AA281797 eneral transcri tion factor IIH; of
a tide 2 44kD subunit
AA282047 ESTs
AA283002 zinc fin er rotein 187
AA283709 cal ain like rotease
AA283902 ESTs
AA284108 Human DNA from chromosome 19-s ecific
cosmid F25965;
Human DNA sequence from clone 71 L16
on chromosome Xp11.
A284109 Contains a probable Zinc Finger protein
(pseudo)gene; an
unknown utative ene; a seudo ene with
hi h similarit to art
AA284371 interleukin 13 rece tor; al ha 1
AA284744 ESTs; Hi hl similar to refoldin subunit
2 M.musculus
AA284784 mitochondrial ribosome rec clin factor
1 5 AA284840 ESTs
AA286844 ESTs
AA287032 ESTs
AA287038 ESTs
AA287546 ESTs
2 0 AA287553 ESTs
AA287556 ESTs; Weakl similar to !!!! ALU CLASS
B WARNING ENTRY !!!!
AA287564 IDN3 rotein
AA291015 CDC7 cell division c cle 7; S. cerevisiae;
homolo -like 1
AA291716 ESTs
2 5 AA291749 estro en rece for 1
AA293656 ESTs
Human DNA sequence from clone 141 H5
on chromosome
A302430 Xq22.1-23. Contains parts of a novel
Chordin LIKE protein with
von Willebrand factor t a C domains.
Contains ESTs; STSs and
AA302809 EST
AA302820 uriner is rece for P2X; Ii and- ated
ion channel; 4
3 0 AA310499 ESTs
AA321890
AA340589 EST
AA340622 ESTs
AA342457 ESTs; Moderatel similar to !!!! ALU
SUBFAMILY SQ WARNING
3 5 AA342828 I co rotein V latelet
AA342864 ESTs
AA342973 ESTs
AA346495 ESTs
I AA347573 fibronectin leucine rich transmembrane
~ protein 2
157
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA347614 ESTs
AA347717 ESTs
AA348913 ESTs
AA349647 EST
AA349773 ESTs
AA350541 ESTs
AA357159 EST
AA357172 ESTs
AA369856 vacuolar rotein sortin 41 east homolo
1 0 AA370132 EST
AA370472 ESTs
AA370867 ESTs
AA377296 ESTs
AA383902 ESTs; Weakl similar to !!!! ALU SUBFAMILY
J WARNING
AA385934 EST; Hi hl similar to redicted usin
Genefinder C.ele ans
AA386255 EST
AA386260 EST
AA386266 ESTs; Weakl similar to M6a H.sa iens
AA398014 ESTs
2 0 AA398222 ESTs
AA398235 ESTs
AA398348 ESTs
AA398482 EST
AA398504 ESTs
2 5 AA398505 ESTs
AA398507 nucleo orin 50kD
AA398523 ESTs
AA398625 ESTs
AA398632 ESTs
3 0 AA398633 ESTs
AA398894 ESTs
AA398895 EST
AA398900 ESTs
AA398904 EST
3 5 AA399122 ESTs; Weakl similar to mitochondria)
citrate trans ort rotein
AA399371 ESTs; Weakl similar to zinc fin er
rotein SALL1 H.sa iens
AA399373 ESTs; Hi hl similar to KIAA0568 rotein
H.sa lens
AA399441 ESTs
AA399636 ESTs
4 0 AA399640 ESTs
AA399680 ESTs
AA400080 ESTs
AA400262 ESTs
158
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA400725 ESTs
AA400748 Homo sa iens mRNA; cDNA DKFZ 434D024
from clone
AA400780 ESTs
AA401631 ESTs
AA401688 ESTs
AA401695 EST
AA402227 ESTs; Weakl similar to TROPOMODULIN
H.sa lens
phosphodiesterase 4A; CAMP-specific
AA402329 (dunce
Droso hila -homolo hos hodiesterase
E2
AA402398 ESTs
1 0 AA402449 EST
AA402468 ESTs
AA403268 ESTs
AA403314 ESTs
AA404229 EST
1 5 AA404260 ESTs
AA404271 lutamate rece tor; ionotro ic; kainate
1
AA405026 ESTs
AA405182 ESTs
AA405237
2 0 AA406061 EST
AA406063 ESTs
AA406070 EST
AA406137 EST
AA406335 ESTs
2 5 AA411801 KIAA0307 ene roduct
AA411804 ESTs
AA411833 ESTs; Hi hl similar to Trad H.sa iens
AA412219 ESTs
AA412259 ESTs
3 0 AA412497 EST
AA412498 ESTs
AA416586 ESTs
AA416867 EST
AA416874 ESTs
3 5 AA421133 ESTs
AA421138 EST
AA422079 ESTs; Weakl similar to RAR-RESPONSIVE
PROTEIN TIG1
AA423837 ESTs
AA424328 ESTs
4 0 AA424339 ESTs
AA424469 ESTs
~ AA424502 ESTs
~
159
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA425004 ESTs
AA425734 ESTs; Weakl similar to h othetical
rotein H.sa lens
AA425887 ESTs
AA426456 ESTs
AA427396 ESTs
AA427555 KIAA0203 ene roduct
AA428218 ESTs
AA428242 transcri tion factor 9 binds GC-rich
se uences
AA428281 EST
1 0 AA428865 EST
AA428994 ESTs
AA429666 ESTs
AA430181 ESTs
AA430184 ATP/GTP-bindin rotein
AA431288 CD3D anti en; delta of a tide TiT3
com lex
AA431293 ESTs
AA431478 ESTs
AA431492 EST
AA431732 EST
2 0 AA432278 ESTs
AA434411 ESTs
AA435512 ESTs
AA435698 ESTs
AA435711 KIAA0712 ene roduct
2 5 AA435815 Clk-associatin RS-c clo hilin
AA435842 ESTs
AA436475 ESTs
AA436489 ESTs
AA442060 ESTs
3 0 AA442079 ESTs
AA443151 ESTs; Weakl similar to weak similarit
with uinone
AA446133 ESTs
AA447145 Homo sa iens KIAA0399 mRNA; artial
cds
AA447398 EST
3 5 AA447643 ESTs
AA447742 d nein; axonemal; heav of a tide 17-like
AA448226
AA448825 EST
AA449444 ESTs
4 0 AA450087 e ulator of Gz-selective rotein si
r nalin
AA450211 EST
AA450244 ESTs
AA452123 ESTs; Weakl similar to T-com lex rotein
1 OA H.sa iens
AA452155 inc fin er rotein 198
z
160
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA452156 EST
AA453036 ESTs; Weakl similar to similar to
mol bdoterin bios nthesis
AA453526 ESTs
AA454085 EST
AA454103 ESTs
AA454642 ESTs
AA454935 nuclear res irato factor 1
AA456323 ESTs
AA457395 ESTs
AA458850
AA459662 ESTs
AA459668 3-h drox isobut I-Coenz me A h drolase
AA459679 ESTs; Weakl similar to The KIAA0191
ene is ex ressed
AA459702 ESTs
AA460017 ESTs; Weakl similar to dia hanous-related
formin M.musculus
AA460324 ESTs
AA461509 ESTs; Weakl similar to utative 150
H.sa iens
AA464414 ESTs
AA464428 ESTs
2 0 AA470084 ESTs
AA476606 ESTs
AA478521 ESTs
AA478523 ESTs; Moderatel similar to !!!! ALU
SUBFAMILY J WARNING
AA479949 RAB2; member RAS onto ene famil
2 5 AA481252 onto ene TC21
AA485351 KIAA1067 rotein
AA487264 ESTs
AA489072 KIAA0870 rotein
AA489630 KIAA0665 ene roduct
3 0 AA490225 ESTs
AA490227 ESTs
AA490255 ESTs
AA490890 ESTs
AA490916 ESTs
3 5 AA490925 a ile s ; ro ressive m oclonic a ile
s ; t a 2 ene; Lafora
AA490955 ESTs; Weakl similar to bullous em
hi oid anti en M.musculus
AA495812 ESTs
AA495824 ESTs
AA496369 ESTs
4 0 AA504125 ESTs
AA521473 SEC10 (S. cerevisiae)-like 1
161
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
AA598440 EST
AA598899 Homo sa iens mRNA; cDNA DKFZ 564D036
from clone
AA599244 KIAA0530 rotein
AA599694 KIAA0133 ene roduct
AA600037 ESTs
AA609135 ESTs
AA609582 katanin 60 ATPase-containin subunit
A 1
AA609684 ESTs
AA609839 4-vitro hen I hos hatase domain and
non-neuronal SNAP25-like
AA609862 RNA-bindin rotein eve with multi )e
s licin
AA620423 EST
AA620747 ESTs
AA621364 ESTs
C20653 ESTs
D20085 ESTs; Weakl similar to KIAA0742 rotein
H.sa iens
D20749 ESTs
D51285 ESTs
D59972 cullin 5
F04112 ESTs
2 0 F13604 ESTs
H01662 ESTs
H05135 ESTs
H12245
H22842 EST
2 5 H30894 ESTs
H43442 leuc I-tRNA s nthetase; mitochondria)
H45996 utative G rotein-cou led rece for
H69281 ESTs
H69485 ESTs
3 0 H69899 ESTs; Moderate) similar to unknown
H.sa lens
H70627 ESTs; Weakl similar to !!!! ALU CLASS
E WARNING ENTRY !!!!
H73050 Rhesus blood rou ; D anti en
H73260 ESTs
H77531 HIR histone cell c cle re ulation
defective; S. cerevisiae
3 5 H80552 EST
H80737 I s I oxidase
H93412 ESTs; Weakl similar to ORF YGR101w
S.cerevisiae
H94892 v-ral simian leukemia viral onto eve
homolo A ras related
H95643 neurotro hit t rosine kinase; rece
tor; t a 1
4 0 H96552 ESTs
H97146 ESTs; Hi hl similar to G rotein-cou
led rece for kinase 6;
H99131 ESTs
162
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
H99462 ribosomal rotein; mitochondrial; L12
H99837 ESTs
N22140 ESTs; Weakl similar to beta-tubulin
H.sa lens
N22197 Sec23-interactin rotein 125
N23756 solute carrier famil 23 nucleobase
traps orters ; member 1
N24134 euka otic translation initiation factor
1A; Y chromosome
N24195 novel centrosomal rotein RanBPM
N26739 DKFZP564B147 rotein
N27098 EST
1 0 N27637 ESTs
N33090 DEAD/H As -Glu-Ala-As /His box of a
tide 19 Db 5; east;
N35967 serinelthreonine kinase 24 Ste20; east
homolo
N38959 cha eronin containin TCP1; subunit
2 beta
N39069 ESTs
1 5 N46441 ESTs
N48270 ESTs
N48365 ESTs
N51316 ESTs
N51499 A kinase PRKA anchor rotein 2
2 0 N53976 ESTs
N54157 ESTs
N54300 ESTs
N54831 ESTs
N59849 ESTs
2 5 N62132 ESTs
N62375 EST
N63138 ESTs
N63172 cell division c cle 42 GTP-bindin rotein;
25kD
novel putative protein similar to YIL091C
N63772 yeast hypothetical 84 kD
rotein from SGA1-KTR7
3 0 N63787 sema domain; immuno lobulin domain
I ; short basic domain;
N68168
N68201 ESTs
N68300 ESTs
N68321 EST
3 5 N69575 EST
N75007 ESTs; Moderatel similar to KIAA1004
rotein H.sa iens
N75542 transcri tion factor 4
O-linked N-acetylglucosamine (GIcNAc)
N90066 transferase
UDP-N-acet I lucosamine: of a tide-N-acet
I lucosamin I
N91246 ESTs
4 0 N92751 ESTs; Weakl similar to MICROTUBULE-ASSOCIATED
N93214 KIAA0318 protein
163
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
N99148 ESTs; Weakl similar to ZINC FINGER
PROTEIN 83 H.sa iens
807876 ESTs; Weakl similar to unknown S.cerevisiae
810865 al ha-feto rotein
811056 ESTs
811488 ESTs
822947 ESTs
823930 ESTs; Hi hl similar to rediabetic
NOD sera-reactive autoanti en
826589 ESTs
837588 RAB2; member RAS onco ene famil -like
837613 Homo sa iens clone 25027 mRNA se uence
838398 Homo sa iens clone 23758 mRNA se uence
839179 ESTs
840923 ESTs
841179 Human mRNA for KIAA0328 ene; artial
cds
1 5 841294 ESTs
842307 earl develo ment re ulator 2 homolo
of of homeotic 2
843189 ESTs
843306 ESTs
844357 ESTs; Weakl similar to cDNA EST EMBL:T01421
comes from
2 0 844519 EST; Moderatel similar to Pro-Pol-dUTPase
of rotein
845088
847948 ESTs
851524 ESTs
854950 ESTs
2 5 855241 ESTs
859585 ESTs
860044 ESTs; Hi hl similar to BETA-CATENIN
H.sa lens
860872 ESTs
866690 ESTs
3 0 867266 exostoses multi 1e -like 1
873588 ESTs
879403 ESTs
887647 ESTs
893622 euka otic translation initiation factor
2; subunit 2 beta; 38kD
3 5 899599 hetero eneous nuclear ribonucleo rotein
U scaffold attachment
899612 ESTs; Moderatel similar to !!!! ALU
SUBFAMILY J WARNING
T02888
T03170 EST
T10465
4 0 T15418 EST
T15597 KIAA0661 ene roduct
T15652 ~ ESTs
164
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
T16898 ash2 absent; small; or homeotic; Droso
hila; homolo -like
T26644 ESTs; Weakl similar to zinc fin er
rotein H.sa iens
T40841 ESTs
T47566
T50116
T50145
T58615 ESTs
T59940 ESTs
T63595 ESTs
T64891
T64924 ESTs
ESTs; Weakly similar to !!!! ALU SUBFAMILY
T64933 SQ WARNING
ENTRY !!!! H.sa lens
T68875
T69027 ESTs
T69924
T70353 ESTs
T79780 ESTs; Weakl similar to CGI-69 rotein
H.sa iens
T79951 ESTs
T80174 ESTs; Moderatel similar to similar
to NEDD-4 H.sa lens
2 0 T80622 ESTs; Weakl similar to envelo a H.sa
lens
T85352 ESTs
T85373 ESTs
T86284 ESTs
T89579 transcri tion factor D -1
2 5 T90360 ESTs
T94328 ESTs
T95590
T97257 ESTs
T97599 ESTs
3 0 T97620 ESTs
T97775 EST
T98152 fibrillin 2 con enital contractural
arachnodact I
W31479 ESTs
W37999 ESTs
3 5 W38240
W40150 chondroitin sulfate roteo I can 6 bamacan
W45435 KIAA0784 rotein
W58202 ESTs
W58344 ESTs
4 0 W58650 ESTs
Human DNA sequence from clone 1189B24
on chromosome
Xq25-26.3. Contains NADH-Ubiquinone
68736 Oxidoreductase MLRO
subunit (EC 1.6.5.3; EC 1.6.99.3; CI-MLRQ);
Tubulin Beta and
Proto-onco ene T rosine- rotein Kinase
FER EC 2.7.1.112;
165
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
W69106 chromobox homolo 3 Droso hila HP1
amma
W69111 ESTs
W69385 nuclear mitotic a aratus rotein 1
W69399 H1 histone famil ; member 0
W69459 sex comb on midle Droso hila -like
1
W72424 S100 calcium-bindin rotein A9 cal
ranulin B
W72724 ESTs
W72834 ESTs
W73955 Homo sa iens chromosome 19; cosmid
826445
W74701 ESTs
W76540 DKFZP564G2022 rotein
W79397 ESTs
W85888 ESTs; Moderatel similar to !!!! ALU
SUBFAMILY SQ WARNING
W86038 ESTs
W86881 ESTs
W87804 ESTs
W88942
W90022 ESTs; Hi hl similar to LECT2 recursor
H.sa lens
W92272 chromodomain helicase DNA bindin rotein
3
2 0 W92764 tumor necrosis factor; al ha-induced
rotein 6
W93040 Homo sa iens aired mesoderm homeo
box 1 PMX1 ; mRNA
W93092 neutral s hin om elinase N-SMase activation
associated factor
W93227 EST
W93523 ESTs
2 5 W93659 ESTs
W94003 ESTs
W94401 ESTs
W94688 erili in
W94787 destrin actin de of merizin factor
3 0 238294 ESTs
238311 ESTs
238465 ESTs
238525 ESTs
238538 ESTs
3 5 238551 ESTs
238783 Ca2+-de endent activator rotein for
secretion
239113 ESTs
239255 YDD19 rotein
239591 EST
4 0 239783 ESTs; Weakl similar to K01 H12.1 C.ele
ans
239920 ESTs; Weakl similar to NADH-CYTOCHROME
B5 REDUCTASE
240166 ESTs
240388 ESTs
166
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Exemplar
Accession Com lete Title UniGeneID
11/29/99
240646 ESTs
241697 ESTs
299349 ESTs
299394 zinc fin er rotein 36 KOX 18
167
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TABLE 4
Exemplar UniGeneID(11/29/
Accession Complete Title 99)
D86425 Homo sapiens mRNA for nidogen-2 Hs.82733
D86983 Human mRNA for KIAA0230 gene; partial Hs.118893
cds
HG1098-HT1098Cystatin D
HG1103-HT1103Guanine Nucleotide-Binding Protein Ral,
Ras-Oncogene Related
HG3342-HT3519Id 1
J03764 plasminogen activator inhibitor; type Hs.82085
I
L06797 chemokine (C-X-C motif); receptor 4 (fusin)Hs.89414
L15388 Human G protein-coupled receptor kinase Hs.211569
(GRKS) mRNA, complete cds
phosphodiesterase 4B; cAMP-specific (dunce
L20971 (Drosophila)-homolog Hs.188
phosphodiesterase E4)
L35545 endothelial cell protein C/activated proteinHs.82353
C receptor
L76380 calcitonin receptor-like Hs.152175
M21305 Human alpha satellite and satellite 3 Hs.247946
junction DNA sequence
M24736 selectin E (endothelial adhesion moleculeHs.89546
1 )
M31166 pentaxin-related gene; rapidly induced Hs.2050
by IL-1 beta
M31551 plasminogen activator inhibitor; type Hs.75716
II (arginine-serpin)
M32334 intercellular adhesion molecule 2 Hs.83733
2 0 M61916 laminin; beta 1 Hs.82124
Human phosphatidylcholine 2-acylhydrolase
M68874 (cPLA2) mRNA, complete
cds
M74719 transcription factor 4 Hs.75356
M92934 connective tissue growth factor Hs.75511
M94856 fatty acid binding protein 5 (psoriasis-associated)Hs.153179
003057 singed (Drosophila)-like (sea urchin fascinHs.118400
homolog like)
003877 EGF-containing fibulin-like extracellularHs.76224
matrix protein 1
018300 damage-specific DNA binding protein 2 Hs.77602
(48kD)
027109 Human prepromultimerin mRNA; complete Hs.32934
cds
031384 guanine nucleotide binding protein 11 Hs.83381
3 0 033053 protein kinase C-like 1 Hs.2499
059423 MAD (mothers against decapentaplegic; Hs.79067
Drosophila) homolog 1
070322 karyopherin (importin) beta 2 Hs.168075
081607 kinase scaffold protein gravin Hs.788
083463 syndecan binding protein (syntenin) Hs.8180
3 5 089942 l ysyl oxidase-like 2 Hs.83354
X04729 Human mRNA for plasminogen activator inhibitor
type 1 N-terminus
X06256 i ntegrin; alpha 5 (fibronectin receptor; Hs.149609
alpha polypeptide)
X07820 matrix metalloproteinase 10 (stromelysin Hs.2258
2)
X54925 matrix metalloproteinase 1 (interstitial Hs.83169
collagenase)
4 0 X54936 placental growth factor; vascular endothelialHs.2894
growth factor-related protein
t yrosine kinase with immunoglobulin and
X60957 epidermal growth factor Hs.78824
homology domains
X67235 hematopoietically expressed homeobox Hs.118651
X67951 proliferation-associated gene A (natural Hs.180909
killer-enhancing factor A)
168
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
X69910 H.sapiens p63 mRNA for transmembrane proteinHs.74368
X79981 cadherin 5; VE-cadherin (vascular epithelium)Hs.76206
218951 caveolin 1; caveolae protein; 22kD Hs.247266
zp61 b6.r1 Stratagene endothelial cell
AA187101 937223 Homo sapiens cDNA
clone IMAGE:624659 5', mRNA sequence
N24990 ESTs Hs.26418
881003 Homo sapiens serine protease mRNA; completeHs.154737
cds
AA025351 ESTs Hs.134797
AA027168 ESTs Hs.10031
AA040465 ESTs Hs.8728
AA045136 ESTs Hs.22575
AA054087 phospholipase A2; group IVC (cytosolic; Hs.18858
calcium-independent)
ESTs; Moderately similar to !!!! ALU SUBFAMILY
AA071089 SC WARNING ENTRY Hs.187932
!!!! [H.sapiens]
AA085918 H.sapiens HUNKI mRNA Hs.247482
AA187490 ESTs Hs.21941
AA227926 ESTs Hs.6682
AA234743 ESTs Hs.22120
AA236559 ESTs; Weakly similar to neuronal thread Hs.8768
protein AD7c-NTP [H.sapiens]
AA292694 ESTs Hs.3807
AA398243 ESTs; Moderately similar to (defline not Hs.21806
available 3694664) [H.sapiens]
2 0 AA406363 ESTs Hs.30822
AA411465 ESTs Hs.8619
AA412284 poliovirus receptor Hs.171844
AA423987 ESTs Hs.7567
AA425309 ESTs Hs.33287
2 5 AA435896 ESTs Hs.18397
AA448238 Homo sapiens mRNA for KIAA0915 protein; Hs.16714
complete cds
AA478778 ESTs Hs.16450
AA621714 ESTs Hs.25338
Human isolate JuSo MUC18 glycoprotein
D51069 mRNA (3' variant); complete Hs.211579
cds
UDP-N-acetyl-alpha-D-galactosamine:polypeptide
3 0 T34527 N-acetylgalactosaminyltransferase 1 (GaINAc-T1Hs.80120
)
U97519 podocalyxin-like Hs.16426
AA127221 ESTs Hs.71059
ESTs; Moderately similar to C-1-TETRAHYDROFOLATE
AA132983 SYNTHASE; Hs.44155
CYTOPLASMIC [H.sapiens]
ESTs; Weakly similar to !!!! ALU SUBFAMILY
AA135606 SB WARNING ENTRY !!!! Hs.189384
[H.sapiens]
3 5 AA156125 ESTs Hs.72116
AA179845 RAB6 interacting; kinesin-like (rabkinesin6)Hs.73625
AA232645 ESTs Hs.42699
F10399 ESTs Hs.14763
H16772 ESTs Hs.31444
40 N39584 ~ ESTs Hs.17404
~
169
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
UDP-N-acetyl-alpha-D-galactosamine:polypeptide
N52006 N-acetylgalactosaminyltransferase 1 (GaINAc-T1Hs.80120
)
N53375 Homer; neuronal immediate early gene; Hs.166146
3
N54067 Homo sapiens mRNA for NIK; partial cds Hs.3628
N64436 ESTs Hs.20813
826892 ESTs Hs.221434
T33637 ESTs Hs.6841
yc20g11.s1 Stratagene lung (#937210) Homo
T57112 sapiens cDNA clone
IMAGE:81284 3', mRNA sequence.
W80763 ESTs; Moderately similar to FK506-bindingHs.3849
protein 65kD [M.musculus]
AA046808 ESTs; Highly similar to 40S RIBOSOMAL Hs.108957
PROTEIN S27 [H.sapiens]
AA253217 ESTs Hs.41271
AA255991 ESTs Hs.175319
AA258138 ESTs Hs.88297
AA426573 ESTs Hs.41135
AA443793 ESTs Hs.94761
AA490588 ESTs Hs.43118
AA496257 ESTs; Weakly similar to (defline not availableHs.72165
3513303) [H.sapiens]
ESTs; Weakly similar to MICROTUBULE-ASSOCIATED
AA609717 PROTEIN 1 B Hs.66048
[H.sapiens]
D59570 ESTs Hs.17132
F13787 ESTs Hs.58596
2 0 H88157 ESTs Hs.41105
H98988 ESTs Hs.42612
N34287 unc5 (C.elegans homology C Hs.44553
N52090 EST Hs.47420
ESTs; Weakly similar to !!!! ALU CLASS
N66845 B WARNING ENTRY !!!! Hs.165411
[H.sapiens]
2 5 N68905 small inducible cytokine A5 (RANTES)
832894 ESTs Hs.45514
861715 ESTs Hs.138237
yi54c08.s1 Soares placenta Nb2HP Homo
sapiens cDNA clone
71234 IMAGE:143054 3' similar to gb~M87908~HUMALNE32
Human carcinoma
cell-derived Alu RNA transcript, (rRNA);
gb:S41458 ROD
yr30g11.s1 Soares fetal liver spleen 1
898105 NFLS Homo sapiens cDNA clone
IMAGE:206852 3', mRNA sequence.
3 0 T97186 small inducible cytokine A5 (RANTES)
ESTs; Moderately similar to !!!! ALU SUBFAMILY
W80814 ! SB WARNING ENTRY
!!! [H.sapiens] s.193700
AA404418 EST Hs.144953
AA405747 ESTs; Moderately similar to HMG-box transcriptionHs.97865
factor [M.musculus]
ESTs; Moderately similar to !!!! ALU SUBFAMILY
AA488687 SQ WARNING ENTRY
! !!! [H.sapiens] s.190307
ESTs; Moderately similar to !!!! ALU SUBFAMILY
3 5 AA599143 SQ WARNING ENTRY
! !!! [H.sapiens]
AA608588 ESTs Hs.193634
170
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
ESTs; Moderately similar to !!!! ALU SUBFAMILY
AA608751 SC WARNING ENTRY Hs.244904
!!!! [H.sapiens]
C13961 EST Hs.210115
D60302 ESTs Hs.108977
H94892 v-ral simian leukemia viral oncogene homologHs.6906
A (ras related)
N93521 transcription factor 4 Hs.241362
N95477 ESTs Hs.102943
ESTs; Weakly similar to !!!! ALU SUBFAMILY
860044 J WARNING ENTRY !!!! Hs.106706
[H.sapiens]
870506 ESTs; Moderately similar to transformation-relatedHs.107159
protein [H.sapiens]
ye20f05.s1 Stratagene lung (#937210) Homo
T91518 sapiens cDNA clone
IMAGE:118305 3' similar to contains Alu
repetitive element;contains
T95333 ESTs; Weakly similar to Strabismus [D.melanogaster]Hs.122730
845630 ESTs; Highly similar to KIAA0372 [H.sapiens]Hs.170098
yg05c07.r1 Soares infant brain 1 NIB Homo
820839 sapiens cDNA clone
IMAGE:31444 5', mRNA sequence.
823858 ESTs; Moderately similar to envelope proteinHs.23986
[H.sapiens]
A1024874 ESTs; Weakly similar to (defline not availableHs.57958
3882257) [H.sapiens]
W26247 U5 snRNP-specific protein (220 kD); orthologHs.6413
of S. cerevisiae Prp8p
AA856990 ESTs Hs.125058
AA136653 ESTs
AA358869 ESTs; Highly similar to SEC13-RELATED Hs.227949
PROTEIN [H.sapiens]
AI123976 ESTs Hs.105689
2 0 AI369384 arylsulfatase D
AA379500 ESTs Hs.193155
849693 ESTs Hs.107708
AA195678 Homo sapiens mRNA for KIAA0465 protein; Hs.108258
partial cds
M30257 vascular cell adhesion molecule 1 Hs.109225
2 5 AA028131 ESTs Hs.110342
M10321 Human von Willebrand factor mRNA, 3' end Hs.110802
J03040 secreted protein; acidic; cysteine-rich Hs.111779
(osteonectin)
M86933 amelogenin (Y chromosome) Hs.1238
AA012933 ubulin-specific chaperone d Hs.241687
t
3 0 AA286710 ymphocyte adaptor protein Hs.13131
l
AA243278 ibosomal protein; mitochondrial; L12 Hs.109059
r
D59711 ESTs Hs.237289
ye36g7.s1 Stratagene lung (#93721 ) Homo
T94452 I sapiens cDNA clone Hs.241207
MAGE:119868 3', mRNA sequence
AA053400 ESTs Hs.241227
Homo sapiens mRNA; cDNA DKFZp58611518
3 5 AA370302 (from clone Hs.21739
DKFZp58611518)
J05008 endothelin 1 Hs.2271
U85193 nuclear factor IlB Hs.33287
AA256153 ESTs Hs.23912
X83107 BMX non-receptor tyrosine kinase Hs.27372
4 0 AA046593 ESTs Hs.28959
~ ~
171
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
AA410480 ESTs Hs.30089
D45304 ESTs Hs.31595
M90657 transmembrane 4 superfamily member 1 Hs.3337
AA010163 upstream regulatory element binding proteinHs.3383
1
AA136353 ESTs Hs.38022
Y07867 pirin Hs.38842
U84573 procollagen-lysine; 2-oxoglutarate 5-dioxygenaseHs.41270
(lysine hydroxylase) 2
X60486 H4 histone family; member G Hs.46423
AA132969 metalloprotease 1 (pitrilysin family) Hs.4812
AA114250 KIAA0512 gene product Hs.48924
F13782 LIM binding domain 2 Hs.4980
ESTs; Weakly similar to IIII ALU SUBFAMILY
AA283035 J WARNING ENTRY IIII Hs.54813
[H.sapiens]
AB002301 Human mRNA for KIAA0303 gene; partial Hs.54985
cds
Sjogren syndrome antigen A2 (60kD; ribonucleoprotein
AA056731 autoantigen Hs.554
SS-A/Ro)
U68019 MAD (mothers against decapentaplegic; Hs.211578
Drosophila) homolog 3
H99198 ESTs; Moderately similar to THYMOSIN BETA-4Hs.56145
[H.sapiens]
AA598702 bone morphogenetic protein 6 Hs.6101
N77151 Homo sapiens mRNA for KIAA0799 protein; Hs.61638
partial cds
AA505133 ESTs Hs.62273
2 0 AB000584 prostate differentiation factor Hs.116577
D12763 interleukin 1 receptor-like 1 Hs.66
AA253193 ESTs Hs.6631
AA432248 ESTs Hs.6738
AA083572 v-ral simian leukemia viral oncogene homologHs.6906
A (ras related)
2 5 AA479713 ESTs Hs.71962
L40395 Homo sapiens clone 23689 mRNA; complete Hs.170001
cds
X52947 gap junction protein; alpha 1; 43kD (connexinHs.74471
43)
W80846 vesicle-associated membrane protein 5 Hs.74669
(myobrevin)
M34539 FK506-binding protein 1A (12kD) Hs.752
3 0 D67029 SEC14 (S. cerevisiae)-like Hs.75232
U09587 glycyl-tRNA synthetase Hs.75280
M85289 Human heparan sulfate proteoglycan (HSPG2)Hs.211573
mRNA, complete cds
D10522 myristoylated alanine-rich protein kinaseHs.75607
C substrate (MARCKS; 80K-L)
W84712 calumenin Hs.7753
3 5 D29992 tissue factor pathway inhibitor 2 Hs.78045
L34657 platelet/endothelial cell adhesion moleculeHs.78146
(CD31 antigen)
S78569 laminin; alpha 4 Hs.78672
D43636 Human mRNA for KIAA0096 gene; partial Hs.79025
cds
U97188 IGF-II mRNA-binding protein 3 Hs.79440
4 0 AA487558 ESTs Hs.8135
M28882 Human MUC18 glycoprotein mRNA, complete Hs.211579
cds
X70683 SRY (sex determining region Y)-box 4 Hs.83484
X14787 ~t hrombospondin 1 Hs.87409
172
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
ESTs; Weakly similar to !!!! ALU CLASS
AA236324 A WARNING ENTRY !!!! Hs.92381
[H.sapiens]
C15324 ESTs Hs.93668
AA452000 ESTs Hs.94030
D83174 collagen-binding protein 2 (colligen 2) Hs.9930
Homo Sapiens gene for thymidylate synthase;
D00596 exons 1; 2; 3; 4; 5; 6; 7; Hs.196351
complete cds
D11428 peripheral myelin protein 22 Hs.103724
D13640 major histocompatibility complex; class Hs.183618
I; C
D14874 adrenomedullin Hs.394
D26129 ribonuclease; RNase A family; 1 (pancreatic)Hs.78224
D28476 thyroid hormone receptor interactor 12 Hs.138617
D86425 Homo sapiens mRNA for nidogen-2 Hs.82733
D86983 Human mRNA for KIAA0230 gene; partial cds Hs.118893
D87953 N-myc downstream regulated Hs.75789
HG1862-HT1897Calmodulin Type I
HG2614-HT2710Collagen, Type Viii, Alpha 1
HG2639-HT2735Single-Stranded Dna-Binding Protein Mssp-1
HG2855-HT2995Heat Shock Protein, 70 Kda (Gb:Y00371 )
HG3044-HT3742Fibronectin, Alt. Splice 1
H 63342-HT3519Id 1
2 0 HG3543-HT3739Insulin-Like Growth Factor 2
HG4069-HT4339Monocyte Chemotactic Protein 1
HG417-HT417Cathepsin B
J03764 plasminogen activator inhibitor; type I Hs.82085
L06797 chemokine (C-X-C motif); receptor 4 (fusin)Hs.89414
L08246 myeloid cell leukemia sequence 1 (BCL2-related)Hs.86386
L12711 t ransketolase (Wernicke-Korsakoff syndrome)Hs.89643
L13977 prolylcarboxypeptidase (angiotensinase Hs.75693
C)
L15388 Human G protein-coupled receptor kinase
(GRK5) mRNA, complete cds
L19871 activating transcription factor 3 Hs.460
3 0 L20859 Human leukemia virus receptor 1 (GLVR1 Hs.78452
) mRNA; complete cds
L42176 f our and a half LIM domains 2 Hs.8302
L49169 Human GOS3 mRNA; complete cds Hs.75678
L76380 calcitonin receptor-like Hs.152175
M15990 v-yes-1 Yamaguchi sarcoma viral oncogene Hs.194148
homolog 1
3 5 M23254 calpain; large polypeptide L2 Hs.76288
M24736 s electin E (endothelial adhesion molecule Hs.89546
1 )
M26576 c ollagen; type IV; alpha 1 Hs.119129
M27396 a sparagine synthetase Hs.75692
M31166 p entaxin-related gene; rapidly induced by Hs.2050
IL-1 beta
Homo sapiens aldehyde dehydrogenase (ALDH1
4 0 M31994 c ) gene, exon 13 and
omplete cds
M32334 i ntercellular adhesion molecule 2 Hs.83733
~ M35878 i nsulin-like growth factor binding protein Hs.77326
3
173
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
M36429 postmeiotic segregation increased 2-like Hs.89672
12
M57730 ephrin-A1 Hs.1624
M57731 GR02 oncogene Hs.75765
M60858 nucleolin Hs.79110
M62994 filamin B; beta (actin-binding protein-278)Hs.81008
Human phosphatidylcholine 2-acylhydrolase
M68874 (cPLA2) mRNA, complete
cds
nuclear factor of kappa light polypeptide
M69043 gene enhancer in B-cells Hs.81328
inhibitor; alpha
M74719 transcription factor 4 Hs.75356
M75126 hexokinase 1 Hs.118625
CD59 antigen p18-20 (antigen identified
M84349 by monoclonal antibodies Hs.119663
16.3A5; EJ16; EJ30; EL32 and 6344)
M92843 zinc finger protein homologous to Zfp-36 Hs.198309
in mouse
M92934 connective tissue growth factor Hs.75511
M93056 protease inhibitor 2 (anti-elastase); monocyte/neutrophilHs.183583
M94856 fatty acid binding protein 5 (psoriasis-associated)Hs.153179
M95787 transgelin Hs.75777
Protein kinase inhibitor [human; neuroblastoma
S76965 cell line SH-SY-5Y; Hs.75209
mRNA; 2147 nt]
S81914 DIFFERENTIATION-DEPENDENT GENE 2 Hs.76095
003057 singed (Drosophila)-like (sea urchin fascinHs.118400
homolog like)
003100 catenin (cadherin-associated protein); Hs.178452
alpha 1 (102kD)
2 0 003877 EGF-containing fibulin-like extracellular Hs.76224
matrix protein 1
008021 nicotinamide N-methyltransferase Hs.76669
014391 myosin IC Hs.82251
031384 guanine nucleotide binding protein 11 Hs.83381
032944 dynein; cytoplasmic; light polypeptide Hs.5120
Human spermidine/spermine N1-acetyltransferase
2 5 040369 (SSAT) gene,
complete cds
041767 Human metargidin precursor mRNA, complete
cds
048959 Homo sapiens myosin light chain kinase Hs.75950
(MLCK) mRNA; complete cds
Human nicotinamide N-methyltransferase
051010 gene, exon 1 and 5' flanking
region
051478 ATPase; Na+/K+ transporting; beta 3 polypeptideHs.76941
Human ovarian cancer downregulated myosin
3 0 053445 heavy chain homolog Hs.15432
(Doc1 ) mRNA; complete cds
059289 cadherin 13; H-cadherin (heart) Hs.63984
059423 MAD (mothers against decapentaplegic; Drosophila)Hs.79067
homolog 1
062015 Homo sapiens Cyr61 mRNA, complete cds
Human hepatitis delta antigen interacting
063825 protein A (dipA) mRNA; Hs.66713
complete cds
3 5 067963 Human lysophospholipase homolog (HU-K5) Hs.6721
mRNA; complete cds
073379 Human cyclin-selective ubiquitin carrier Hs.93002
protein mRNA; complete cds
073824 eukaryotic translation initiation factor Hs.183684
4 gamma; 2
077604 microsomal glutathione S-transferase 2 Hs.81874
~ U81607 kinase scaffold protein gravin Hs.788
174
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
089942 lysyl oxidase-like 2 Hs.83354
X04412 gelsolin (amyloidosis; Finnish type) Hs.80562
X06985 heme oxygenase (decycling) 1 Hs.75967
X07820 matrix metalloproteinase 10 (stromelysin Hs.2258
2)
X12876 keratin 18 Hs.65114
X15729 DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptideHs.76053
5 (RNA helicase; 68kD)
X52541 early growth response 1 Hs.738
X53416 filamin A; alpha (actin-binding protein-280)Hs.76279
X54489 GR01 oncogene (melanoma growth stimulatingHs.789
activity; alpha)
X54925 matrix metalloproteinase 1 (interstitial Hs.83169
collagenase)
X57206 inositol 1;4;5-trisphosphate 3-kinase B Hs.78877
X59798 cyclin D1 (PRAD1: parathyroid adenomatosisHs.82932
1)
tyrosine kinase with immunoglobulin and
X60957 epidermal growth factor Hs.78824
homology domains
X65965 H.sapiens SOD-2 gene for manganese superoxide
dismutase
X69111 inhibitor of DNA binding 3; dominant negativeHs.76884
helix-loop-helix protein
X70940 eukaryotic translation elongation factor Hs.2642
1 alpha 2
X87838 catenin (cadherin-associated protein); Hs.171271
beta 1 (88kD)
X91247 thioredoxin reductase 1 Hs.13046
X97748 H.sapiens PTX3 gene promotor region
2 0 Y00815 protein tyrosine phosphatase; receptor Hs.75216
type; F
AA303711 ephrin-B1 Hs.144700
L44538 ESTs Hs.156044
AA025351 ESTs Hs.134797
AA027050 ESTs Hs.31189
2 5 AA029462 ESTs Hs.17235
AA045136 ESTs Hs.22575
AA047437 ESTs Hs.22968
AA054087 phospholipase A2; group IVC (cytosolic; Hs.18858
calcium-independent)
ESTs; Moderately similar to !!!! ALU SUBFAMILY
AA071089 SC WARNING ENTRY
!~!! [H.sapiens] s.187932
3 0 AA156450 ESTs; Weakly similar to Similar to Rat Hs.8982
trg gene product [C.elegans]
AA187490 ESTs Hs.21941
ESTs; Moderately similar to PROBABLE G
AA195031 PROTEIN-COUPLED Hs.9305
RECEPTOR APJ [H.sapiens]
AA205724 ESTs Hs.10119
AA227926 ESTs Hs.6682
3 5 AA227986 ESTs Hs.25329
AA234743 ESTs Hs.22120
AA253216 ESTs Hs.22283
AA256210 oncomodulin Hs.199134
AA256268 ESTs Hs.10283
4 0 AA279397 ESTs; Moderately similar to fibronectin Hs.25001
[H.sapiens]
ESTs; Moderately similar to !!!! ALU SUBFAMILY
AA292379 SO WARNING ENTRY Hs.20340
! !!! H.sa lens
175
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
AA292717 ESTs; Weakly similar to JM2 [H.sapiens] Hs.7891
AA346551 ESTs Hs.23457
AA400292 ESTs Hs.23786
AA404338 ESTs Hs.21812
AA412284 poliovirus receptor Hs.171844
AA423987 ESTs Hs.7567
AA428594 ESTs Hs.21321
AA430108 ESTs Hs.6019
AA431462 ESTs Hs.28329
ESTs; Weakly similar to CAMP-DEPENDENT
AA431470 PROTEIN KINASE Hs.3407
INHIBITOR; MUSCLE/BRAIN FORM [H.sapiens]
AA443756 ESTs; Moderately similar to (defline not Hs.6673
available 4105275) [H.sapiens]
AA449479 ESTs; Highly similar to (defline not availableHs.5216
5106787) [H.sapiens]
AA459916 bradykinin receptor B2 Hs.25021
AA465226 ESTs Hs.28631
AA478778 ESTs Hs.16450
AA479037 ESTs Hs.7961
AA482597 ESTs; Highly similar to (defline not availableHs.26054
4704739) [H.sapiens]
AA487561 ESTs; Highly similar to RAS-RELATED PROTEINHs.9813
RAB-1A [H.sapiens]
AA489245 ESTs; Weakly similar to sperm specific Hs.5682
protein [H.sapiens]
2 0 AA504110 ESTs Hs.18063
ESTs; Highly similar to SERINE/THREONINE
AA520989 PROTEIN Hs.9195
PHOSPHATASE PP1-BETA CATALYTIC SUBUNIT
[H.sapiens]
AA599434 ESTs Hs.25035
AA608649 Homo sapiens clone 23742 mRNA; partial Hs.6354
cds
AA609519 ESTs Hs.26458
Human isolate JuSo MUC18 glycoprotein
2 5 D51069 mRNA (3' variant); complete Hs.185718
cds
097519 podocalyxin-like Hs.16426
W28391 proliferation-associated 2G4; 38kD Hs.5181
Homo sapiens mRNA; cDNA DKFZp564F053 (from
AA035638 clone Hs.71968
DKFZp564F053)
AA083514 ESTs Hs.68301
3 0 AA121315 ESTs Hs.70823
AA147186 ESTs Hs.92387
AA156125 ESTs Hs.72116
AA188932 ESTs Hs.85640
AA219653 ESTs Hs.87125
3 5 AA232645 ESTs Hs.42699
F10078 ESTs Hs.13233
H48032 ESTs Hs.9645
H82117 ESTs Hs.28043
N39584 ESTs Hs.17404
4 0 N54067 Homo sapiens mRNA for NIK; partial cds Hs.3628
~ N59858 ~ ESTs
Hs.33032
176
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
N90933 ESTs Hs.4867
ESTs; Moderately similar to !!!! ALU CLASS
N93764 C WARNING ENTRY !!!! Hs.10175
[H.sapiens]
826124 ESTs Hs.24024
827957 ESTs Hs.24230
855470 ESTs; Moderately similar to K02E10.2 [C.elegans]Hs.11067
ESTs; Highly similar to vacuolar protein
T16550 sorting homolog h-vps45 Hs.6650
[H.sapiens]
T26674 ESTs; Weakly similar to neuronal thread Hs.6966
protein AD7c-NTP [H.sapiens]
yc20g11.s1 Stratagene lung (#937210) Homo
T57112 sapiens cDNA clone Hs.8881
IMAGE:81284 3', mRNA sequence.
T88700 ESTs Hs.173374
T90527 ESTs Hs.7890
W42789 ESTs Hs.31446
W60002 plastin 3 (T isoform) Hs.4114
W78175 ESTs Hs.17901
W84768 ESTs Hs.141742
W94427 ESTs; Weakly similar to Na;K-ATPase gammaHs.3807
subunit [H.sapiens]
AA253217 ESTs Hs.41271
AA426573 ESTs Hs.41135
AA432374 ESTs Hs.48029
AA446622 ESTs Hs.74313
2 0 AA478771 ESTs Hs.50841
AA482594 ESTs Hs.62684
AA490588 ESTs Hs.43118
D59570 ESTs Hs.17132
H88157 ESTs Hs.41105
H94648 ESTs Hs.41995
H97538 ESTs ' Hs.42392
H98670 ESTs; Weakly similar to (defline not availableHs.49753
4884081 ) [H.sapiens]
ESTs; Moderately similar to !!!! ALU SUBFAMILY
N22107 SC WARNING ENTRY Hs.172241
!!!! [H.sapiens]
W38197 Accession not listed in Genbank
ESTs; Moderately similar to !!!! ALU SUBFAMILY
3 0 W80814 SB WARNING ENTRY Hs.196785
!!!! [H.sapiens]
AA287347 ESTs Hs.105088
AA402799 ESTs Hs.182538
AA404418 EST Hs.144953
AA425107 ESTs Hs.97016
ESTs; Moderately similar to !!!! ALU SUBFAMILY
3 5 AA425435 J WARNING ENTRY Hs.98438
! !!! [H.sapiens]
AA442872 ESTs Hs.110771
ESTs; Moderately similar to !!!! ALU SUBFAMILY
AA452860 SP WARNING ENTRY Hs.197214
! !!! [H.sapiens]
ESTs; Moderately similar to !!!! ALU SUBFAMILY
AA488687 SQ WARNING ENTRY Hs.190307
! !!! [H.sapiens]
AA599674 ESTs; Weakly similar to ORF [D.melanogaster]Hs.108115
177
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
F13673 ESTs Hs.99769
H99093 DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptideHs.6179
(72kD)
yw35g11.s1 Morton Fetal Cochlea Homo Sapiens
N22495 cDNA clone Hs.102415
IMAGE:254276 3', mRNA sequence.
N23031 myosin; heavy polypeptide 7; cardiac muscle;Hs.929
beta
815740 carbohydrate (chondroitin 6/keratan) sulfotransferaseHs.104576
1
839610 calpain; large polypeptide L2 Hs.76288
W45560 ESTs Hs.102541
239833 H.sapiens mRNA for Rho6 protein Hs.124940
240583 ESTs Hs.101259
1 0 AA825437 ESTs
Homo sapiens mRNA; cDNA DKFZp564F053 (from
866613 clone
DKFZp564F053)
AA868063 carbohydrate (chondroitin 6/keratan) sulfotransferase
1
z116d08.r1 Soares_pregnant uterus NbHPU
AA128075 Homo sapiens cDNA clone
IMAGE:502095 5', mRNA sequence.
N66570 ESTs
A1051390 ESTs
AA627122 ESTs
X02761 fibronectin 1 Hs.118162
AF010193 MAD (mothers against decapentaplegic; Hs.100602
Drosophila) homolog 7
ESTs; Highly similar to the KIAA0195 gene
AA149044 is expressed ubiquitously. Hs.10086
[H.sapiens]
solute carrier family 9 (sodium/hydrogen
2 0 U82108 exchanger); isoform 3 Hs.101813
regulatory factor 2
D78676 ESTs; Moderately similar to (defline not Hs.105509
available 4529890) (H.sapiens]
L35240 enigma (LIM domain protein) Hs.102948
AA598737 lactate dehydrogenase B Hs.180414
869417 ESTs Hs.107055
ESTs; Weakly similar to Human pre-mRNA
2 5 AA232837 cleavage factor I 68 kDa Hs.107125
subunit [H.sapiens]
N72695 ESTs Hs.108557
M30257 vascular cell adhesion molecule 1 Hs.109225
M96843 inhibitor of DNA binding 2; dominant negativeHs.109617
helix-loop-helix protein
X68277 dual specificity phosphatase 1 Hs.171695
3 0 AA292440 myeloid differentiation primary response Hs.110571
J03040 secreted protein; acidic; cysteine-rich Hs.111779
(osteonectin)
AA228107 ESTs Hs.54642
AA449789 connective tissue growth factor Hs.75511
W01367 ESTs Hs.170980
3 5 AA610116 ESTs; Highly similar to (defline not availableHs.11663
4325180) [H.sapiens]
Homo Sapiens mRNA; cDNA DKFZp564F053 (from
AA258308 clone Hs.165618
DKFZp564F053)
AA460273 Homo Sapiens mRNA for KIAA0517 protein; Hs.12372
partial cds
AA286710 lymphocyte adaptor protein Hs.13131
T68873 metallothionein 1 L Hs.143289
178
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
D63476 PAK-interacting exchange factor beta Hs.172813
M62403 insulin-like growth factor-binding proteinHs.1516
4
X55740 5' nucleotidase (CD73) Hs.153952
L10284 calnexin Hs.155560
AA243278 ribosomal protein; mitochondrial; L12 Hs.109059
AA430032 pituitary tumor-transforming 1 Hs.159626
H16402 ESTs Hs.17121
D59711 ESTs Hs.17132
ye36g7.s1 Stratagene lung (#93721 ) Homo
T94452 sapiens cDNA clone
IMAGE:119868 3', mRNA sequence
AA431571 ESTs Hs.17894
879356 Homo Sapiens mRNA for KIAA0544 protein; Hs.19280
partial cds
AA280375 ESTs Hs.19928
249269 small inducible cytokine subfamily A (Cys-Cys);Hs.20144
member 14
241740 ESTs Hs.24462
AA121543 Homo sapiens mRNA for KIAA0758 protein; Hs.22039
partial cds
J05008 endothelin 1 Hs.2271
AA101878 ESTs Hs.22793
T35341 ESTs; Highly similar to (defline not availableHs.22880
4519883) [H.sapiens]
N87590 ESTs Hs.23037
2 0 AA256153 ESTs Hs.23912
W74533 Homo sapiens mRNA for KIAA0786 protein; Hs.24212
partial cds
U25997 stanniocalcin Hs.25590
V01512 v-fos FBJ murine osteosarcoma viral oncogeneHs.25647
homolog
X56681 jun D proto-oncogene Hs.2780
2 5 AA161292 interferon; alpha-inducible protein 27 Hs.2867
AA491465 ESTs Hs.28792
AA046593 ESTs Hs.28959
D50914 Human mRNA for KIAA0124 gene; partial Hs.30736
cds
D45304 ESTs Hs.31595
3 0 M90657 transmembrane 4 supertamily member 1 Hs.3337
W69127 ESTs; Weakly similar to zinc finger proteinHs.3449
ZNF191 [H.sapiens]
AA316186 ESTs; Highly similar to (defline not availableHs.34549
4262136) [H.sapiens]
AA384503 ESTs Hs.179260
AA136353 ESTs Hs.38022
ESTs; Weakly similar to !!!! ALU SUBFAMILY
3 5 AA044755 SX WARNING ENTRY !!!! Hs.173705
[H.sapiens]
U84573 procollagen-lysine; 2-oxoglutarate 5-dioxygenaseHs.41270
(lysine hydroxylase) 2
AA058911 ESTs; Weakly similar to membrane glycoproteinHs.4193
[M.musculus]
AA620962 dynein; cytoplasmic; light intermediate Hs.44251
polypeptide 2
AA285290 small EDRK-rich factor 2 Hs.44499
4 0 X60486 H4 histone family; member G Hs.46423
831641 ESTs Hs.197148
AA489190 ESTs Hs.48320
F13782 ~ LIM binding domain 2 Hs.4980
179
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
AA257993 Janus kinase 1 (a protein tyrosine kinase)Hs.50651
M24283 intercellular adhesion molecule 1 (CD54); Hs.168383
human rhinovirus receptor
ESTs; Weakly similar to PIM-1 PROTO-ONCOGENE
AA443114 SERINE/THREONINE-PROTEIN KINASE [H.sapiens]Hs.5326
T35289 casein kinase 1; alpha 1 Hs.195206
N23817 Homo sapiens clone 23675 mRNA sequence Hs.5807
AA047151 ESTs Hs.5897
N77151 Homo sapiens mRNA for KIAA0799 protein; Hs.61638
partial cds
AA480074 ESTs Hs.62206
Y00787 interleukin 8 Hs.624
T99789 ESTs Hs.64313
W84341 tissue inhibitor of metalloproteinase 2 Hs.6441
L09209 amyloid beta (A4) precursor-like protein Hs.64797
2
D12763 interleukin 1 receptor-like 1 Hs.66
T16484 ESTs Hs.6607
AA253193 ESTs Hs.6631
AA432248 ESTs Hs.6738
X82200 stimulated trans-acting factor (50 kDa) Hs.68054
AA083572 v-ral simian leukemia viral oncogene homologHs.6906
A (ras related)
L00352 low density lipoprotein receptor (familialHs.181182
hypercholesterolemia)
2 0 N75791 ESTs Hs.7153
X57579 H.sapiens activin beta-A subunit (exon
2)
cytochrome P450; subfamily I (aromatic
X02612 compound-inducible); Hs.72912
polypeptide 1
H44631 immediate early protein Hs.737
AA090257 superoxide dismutase 2; mitochondria) Hs.177781
2 5 X83703 H.sapiens mRNA for cytokine inducible nuclearHs.74019
protein
L40395 Homo sapiens clone 23689 mRNA; complete Hs.170001
cds
AA227913 ESTs Hs.198456
X52947 gap junction protein; alpha 1; 43kD (connexinHs.74471
43)
M11313 alpha-2-macroglobulin Hs.74561
3 0 L14837 tight junction protein 1 (zona occludens Hs.74614
1 )
M60721 Human homeobox gene, complete cds
D90209 activating transcription factor 4 (tax-responsiveHs.181243
enhancer element B67)
yc28e12.s1 Stratagene liver (#937224) Homo
T67986 sapiens cDNA clone Hs.75106
IMAGE:82030 3' similar to gb:X14723 CLUSTERIN
PRECURSOR
AA148318 Human mRNA for KIAA0069 gene; partial cds Hs.75249
3 5 097105 dihydropyrimidinase-like 2 Hs.173381
T25747 H.sapiens OZF mRNA Hs.75471
K02574 Accession not listed in Genbank
tyrosine 3-monooxygenase/tryptophan 5-monooxygenase
D78577 activation Hs.75544
protein; eta polypeptide
X53331 matrix Gla protein Hs.75742
40 S73591 upregulated by 1;25-dihydroxyvitamin D-3 Hs.179526
X95735 Zyxin ___. Hs.75873
I
180
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Exemplar UniGeneID(11/29/
Accession Complete Title 99)
L16862 G protein-coupled receptor kinase 6 Hs.76297
044975 Homo Sapiens Kruppel-like zinc finger Hs.76526
protein Zf9 mRNA; complete cds
M97796 inhibitor of DNA binding 2; dominant negativeHs.180919
helix-loop-helix protein
086782 26S proteasome-associated pad1 homolog Hs.178761
AA099391 ESTs Hs.77310
M19267 tropomyosin 1 (alpha) Hs.77899
D29992 tissue factor pathway inhibitor 2 Hs.78045
L19314 phosphorylase kinase; beta Hs.195217
S78569 laminin; alpha 4 Hs.78672
Human cysteine-rich fibroblast growth
028811 factor receptor (CFR-1 ) mRNA,
complete cds
L77886 protein tyrosine phosphatase; receptor Hs.79005
type; K
C14407 neuronal tissue-enriched acidic protein Hs.79516
diphtheria toxin receptor (heparin-binding
M60278 epidermal growth factor-like Hs.799
growth factor)
881509 splicing factor; arginine/serine-rich Hs.184571
11
AA487558 ESTs Hs.8135
D86962 KIAA0207 gene product Hs.81875
AA478971 disabled (Drosophila) homolog 2 (mitogen-responsiveHs.81988
phosphoprotein)
D50683 transforming growth factor; beta receptorHs.82028
II (70-80kD)
056637 capping protein (actin filament) muscle Hs.184270
Z-line; alpha 1
2 0 M61199 Human cleavage signal 1 protein mRNA; Hs.82767
complete cds
M28882 Human MUC18 glycoprotein mRNA, complete
cds
X15183 CDW52 antigen (CAMPATH-1 antigen) Hs.180532
S53911 CD34 Hs.85289
020734 Human transcription factor junB QunB) Hs.198951
gene; 5' region and complete cds
prostaglandin-endoperoxide synthase 2
2 5 D28235 (prostaglandin G/H synthase Hs.92309
and cyclooxygenase)
ESTs; Weakly similar to !!!! ALU CLASS
AA236324 A WARNING ENTRY !!!! Hs.92381
[H.sapiens]
Homo Sapiens mRNA for DEPP (decidual protein
AA148923 induced by Hs.93675
progesterone); complete cds
AA174183 ESTs Hs.93872
ESTs; Weakly similar to !!!! ALU CLASS
AA456311 A WARNING ENTRY !!!! Hs.93961
[H.sapiens]
3 0 L08069 heat shock protein; DNAJ-like 2 Hs.94
AA452000 ESTs Hs.94030
AA282140 ESTs Hs.9587
J02854 myosin regulatory light chain 2; smooth Hs.9615
muscle isoform
~ AA442054 phospholipase C; gamma 1 (formerly subtypeHs.993
~ 148)
181
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TABLE 5
AccessionUniGeneIDTitle Gene Eos
# #
AA426573Hs.41135ESTs; Moderately similar to endomucin AAA9
[M.musculus]
D58024 Hs.57958ESTs; Weakly similar to KIAA0768 AAA8
protein [H.sapiens]
endothelial differentiation; sphingolipid
M31210 Hs.154210G-protein-coupled EDG1 AAA7
receptor; 1
X06256 Hs.149609integrin; alpha 5 (fibronectin ITGA5 AAB1
receptor; alpha polypeptide)
L20859 Hs.78452solute carrier family 20 (phosphateSLC20A1AAB3
transporter); member 1
X07820 Hs.2258matrix metalloproteinase 10 (stromelysinMMP10 AAB4
2)
AA234743Hs.22120ESTs AAB5
U97519 Hs.16426podocalyxin-like PODXL AAB6
U03877 Hs.76224EGF-containing fibulin-like extracellularEFEMP1AAB8
matrix protein 1
M28882 Hs.211579melanoma adhesion molecule MCAM AAB9
X54925 Hs.83169matrix metalloproteinase 1 (interstitialMMP1 AAC1
collagenase)
AA045136Hs.22575ESTs AAC2
AA423987Hs.7567ESTs AAC3
AA234743Hs.22120ESTs AAC4
ESTs; Moderately similar to hedgehog-interacting
AA156125Hs.72116protein AAC5
(M.musculus]
AA025351Hs.134797ESTs AAC6
AA432248Hs.6738ESTs AAC7
2 0 AA227926Hs.6682ESTs AAC8
AA187490Hs.21941ESTs AAD1
~ AA232645Hs.42699ESTs AAD2
~ ~
182
