A METHOD FOR THE PROPHYLAXIS AND/OR TREATMENT OF MEDICAL DISORDERS

METHODE DE PROPHYLAXIE ET/OU DE TRAITEMENT DE TROUBLES CLINIQUES

English Abstract

The present invention relates generally to a method for the prophylaxis and/or
treatment of skin disorders, and in particular proliferative and/or
inflammatory skin disorders, and to genetic molecules useful for same. The
present invention is particularly directed to genetic molecules capable of
modulating growth factor interaction with its receptor on epidermal
keratinocytes to inhibit, reduce or otherwise decrease stimulation of this
layer of cells. The present invention contemplates, in a most preferred
embodiment, a method for the prophylaxis and/or treatment of psoriasis.

French Abstract

La présente invention concerne généralement une méthode de prophylaxie et/ou de traitement de troubles cutanés, et particulièrement de troubles cutanés inflammatoires et/ou proliférants, et des molécules génétiques utiles dans ces cas. Cette invention a notamment trait à des molécules génétiques capables de moduler l'interaction du facteur de croissance avec son récepteur sur des kératinocytes épidermiques pour inhiber, réduire ou diminuer la stimulation de cette couche des cellules. Dans un mode de réalisation préféré, elle porte sur une méthode de prophylaxie et/ou de traitement du psoriasis.

Claims

-113-

CLAIMS:

1. A method for ameliorating the effects of a proliferative and/or
inflammatory
skin disorder in a mammal, said method comprising contacting the proliferating
and/or
inflamed skin or skin capable of proliferation and/or inflammation with an
effective
amount of a nucleic acid molecule selected from the group consisting of 5'-
UCCGGAGCCAGACUU-3' (SEQ ID NO:12); 5'-CACAGUUGCUGCAAG-3' (SEQ ID
NO:13); 5'-UCUCCGCUUCCUUUC-3' (SEQ ID NO:14); 5'-AGCCCCCACAGCGAG-
3' (SEQ ID NO:15); 5'-GCCUUGGAGAUGAGC-3' (SEQ ID NO:16); 5'-
UAACAGAGGUCAGCA-3' (SEQ ID NO:17); 5'-GGAUCAGGGACCAGU-3' (SEQ ID
NO:18); 5'-CGGCAAGCUACACAG-5' (SEQ ID NO:19); 5'-GGCAGGCAGGCACAC-
3' (SEQ ID NO:20) or chemical analogue of any one of said nucleic acid
molecules
wherein said nucleic acid molecule or its chemical analogue is capable of
inhibiting or
otherwise reducing growth factor mediated cell proliferation and/or
inflammation and/or
other medical disorders.
2. A method according to Claim 1 wherein the mammal is a human.
3. A method according to Claim 1 or 2 wherein cell proliferation and/or
inflammation is mediated by at least one of insulin-like growth factor I (IGF-
I),
keratinocyte growth factor (KGF), transforming growth factor-.alpha.
(TGF.alpha.), tumour necrosis
factor-.alpha. (TNF.alpha.), interleukin (IL) -1 (IL-1), IL-4, IL-6, IL-8
and/or basic fibroblast growth
factor (bFGF).
4. A method according to Claim 3 wherein cell proliferation and/or
inflammation is mediated by IGF-I.
5. A method according to Claim 1 or 2 wherein the proliferative or
inflammatory skin disorder is psoriasis, eczema, ichthyosis, pityriasis,
rubra, pilaris,
serborrhoea, keloids, keratosis, neoplasias, scleroderma, warts, benign
growths or cancers
of the skin.



-114-

6. A method according to Claim 5 wherein the skin condition is psoriasis.
7. A method according to Claim 1 or 4 or 6 wherein the nucleic acid molecule
is 5'-UCCGGAGCCAGACUU-3' (SEQ ID NO:12) or chemical analogue thereof.
8. A method according to Claim 1 or 4 or 6 wherein the nucleic acid molecule
is 5'-CACAGUUGCUGCAAG-3' (SEQ ID NO:13) or chemical analogue thereof.
9. A method according to Claim 1 or 4 or 6 wherein the nucleic acid molecule
is 5'-UCUCCGCUUCCUUUC-3' (SEQ ID NO:14) or chemical analogue thereof.
10. A method according to Claim 1 or 4 or 6 wherein the nucleic acid molecule
is 5'-AGCCCCCACAGCGAG-3' (SEQ ID NO:15) or chemical analogue thereof.
11. A method according to Claim 1 or 4 or 6 wherein the nucleic acid molecule
is 5'-GCCUUGGAGAUGAGC-3' (SEQ ID NO:16) or chemical analogue thereof.
12. A method according to Claim 1 or 4 or 6 wherein the nucleic acid molecule
is 5'-UAACAGAGGUCAGCA-3' (SEQ 1D NO:17) or chemical analogue thereof.
13. A method according to Claim 1 or 4 or 6 wherein the nucleic acid molecule
is 5'-GGAUCAGGGACCAGU-3' (SEQ 1D NO:18) or chemical analogue thereof.
14. A method according to Claim 1 or 4 or 6 wherein the nucleic acid molecule
is 5'-CGGCAAGCUACACAG-5' (SEQ ID NO:19) or chemical analogue thereof.
15. A method according to Claim 1 or 4 or 6 wherein the nucleic acid molecule
is 5'-GGCAGGCAGGCACAC-3' (SEQ ID NO:20) or chemical analogue thereof.



-115-

16. A nucleic acid molecule comprising at least about 15 nucleotides capable
of
hybridizing to or forming a heteroduplex or otherwise interacting with a
complementary
form of SEQID ID NO:12 to SEQ ID NO:20 inclusive.
17. A nucleic acid molecule comprising at least about 15 nucleotides capable
of
hybridizing to or forming a heteroduplex or otherwise interacting with a
complementary
form of SEQ ID NO:12 to SEQ ID NO:20 inclusive.
18. A nucleic acid molecule comprising at least about 15 nucleotides capable
of
hybridizing to or forming a heteroduplex or otherwise interacting with a
complementary
form of SEQ ID NO:12 to SEQ ID NO:14 or SEQ ID NO:20 inclusive.
19. A method of ameliorating the effects of psoriasis in a mammal, said method
comprising contacting proliferating skin or skin capable of proliferation with
an effective
amount of one or more nucleic acid molecules or chemical analogues thereof
capable of
inhibiting or otherwise reducing IGF-I mediated cell proliferation wherein
said one or
more molecules comprises a polynucleotide selected from the group consisting
of 5'-
UCCGGAGCCAGACUU-3' (SEQ ID NO:12); 5'-CACAGUUGCUGCAAG-3' (SEQ ID
NO:13); 5'-UCUCCGCUUCCUUUC-3' (SEQ ID NO:14); 5'-AGCCCCCACAGCGAG-
3' (SEQ ID NO:15); 5'-GCCUUGGAGAUGAGC-3' (SEQ ID NO:16); 5'-
UAACAGAGGUCAGCA-3' (SEQ ID NO:17); 5'-GGAUCAGGGACCAGU-3' (SEQ ID
NO:18); 5'-CGGCAAGCUACACAG-5' (SEQ ID NO:19); 5'-GGCAGGCAGGCACAC-
3' (SEQ ID NO:20) or chemical analogue of any one of said nucleic acid
molecules which
is capable of interacting with mRNA directed from an IGF-I gene, an IGF-I
receptor gene
or a gene encoding an IGFBP.
20. A method according to Claim 19 wherein the mammal is a human.
21. A method according to Claim 19 or 20 wherein the nucleic acid molecule is
5'-UCCGGAGCCAGACUU-3' (SEQ ID NO:12) or chemical analogue thereof.



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22. A method according to Claim 19 or 20 wherein the nucleic acid molecule is
5'-CACAGUUGCUGCAAG-3' (SEQ ID NO:13) or chemical analogue thereof.
23. A method according to Claim 19 or 20 wherein the nucleic acid molecule is
5'-UCUCCGCUUCCUUUC-3' (SEQ ID NO:14) or chemical analogue thereof.
24. A method according to Claim 19 or 20 wherein the nucleic acid molecule is
5'-AGCCCCCACAGCGAG-3' (SEQ ID NO:15) or chemical analogue thereof.
25. A method according to Claim 19 or 20 wherein the nucleic acid molecule is
5'-GCCUUGGAGAUGAGC-3' (SEQ ID NO:16) or chemical analogue thereof.
26. A method according to Claim 19 or 20 wherein the nucleic acid molecule is
5'-UAACAGAGGUCAGCA-3' (SEQ ID NO:17) or chemical analogue thereof.
27. A method according to Claim 19 or 20 wherein the nucleic acid molecule is
5'-GGAUCAGGGACCAGU-3' (SEQ ID NO:18) or chemical analogue thereof.
28. A method according to Claim 19 or 20 wherein the nucleic acid molecule is
5'-CGGCAAGCUACACAG-5' (SEQ ID NO:19) or chemical analogue thereof.
29. A method according to Claim 19 or 20 wherein the nucleic acid molecule is
5'-GGCAGGCAGGCACAC-3' (SEQ ID NO:20) or chemical analogue thereof.
30. A composition comprising a nucleic acid molecule capable of inhibiting or
otherwise reducing IGF-I mediated cell proliferation, said composition
comprising a
nucleic acid molecule selected from the group consisting of 5'-UCCGGAGCCAGACUU-

3' (SEQ ID NO:12); 5'-CACAGUUGCUGCAAG-3' (SEQ ID NO:13); 5'-
UCUCCGCUUCCUUUC-3' (SEQ ID NO:14); 5'-AGCCCCCACAGCGAG-3' (SEQ ID
NO:15); 5'-GCCUUGGAGAUGAGC-3' (SEQ ID NO:16); 5'-UAACAGAGGUCAGCA-
3' (SEQ ID NO:17); 5'-GGAUCAGGGACCAGU-3' (SEQ ID NO:18); 5'-


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CGGCAAGCUACACAG-5' (SEQ ID NO:19); 5'-GGCAGGCAGGCACAC-3' (SEQ ID
NO:20) or chemical analogue of any one of said nucleic acid molecules, said
composition
further comprising one or more pharmaceutically acceptable carriers and/or
diluents.
31. A method according to Claim 30 wherein the mammal is a human.
32. A method according to Claim 30 or 31 wherein the nucleic acid molecule is
5'-UCCGGAGCCAGACUU-3' (SEQ ID NO:12) or chemical analogue thereof.
33. A method according to Claim 30 or 31 wherein the nucleic acid molecule is
5'-CACAGUUGCUGCAAG-3' (SEQ ID NO:13) or chemical analogue thereof.
34. A method according to Claim 30 or 31 wherein the nucleic acid molecule is
5'-UCUCCGCUUCCUUUC-3' (SEQ ID NO:14) or chemical analogue thereof.
35. A method according to Claim 30 or 31 wherein the nucleic acid molecule is
5'-AGCCCCCACAGCGAG-3' (SEQ ID NO:15) or chemical analogue thereof.
36. A method according to Claim 30 or 31 wherein the nucleic acid molecule is
5'-GCCUUGGAGAUGAGC-3' (SEQ ID NO:16) or chemical analogue thereof.
37. A method according to Claim 30 or 31 wherein the nucleic acid molecule is
5'-UAACAGAGGUCAGCA-3' (SEQ ID NO:17) or chemical analogue thereof.
38. A method according to Claim 30 or 31 wherein the nucleic acid molecule is
5'-GGAUCAGGGACCAGU-3' (SEQ ID NO:18) or chemical analogue thereof.
39. A method according to Claim 30 or 31 wherein the nucleic acid molecule is
5'-CGGCAAGCUACACAG-5' (SEQ ID NO:19) or chemical analogue thereof.



-118-

40. A method according to Claim 30 or 31 wherein the nucleic acid molecule is
5'-GGCAGGCAGGCACAC-3' (SEQ ID NO:20) or chemical analogue thereof.

Description

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A METHOD FOR THE PROPHYLAXIS AND/OR TREATMENT OF
MEDICAL DISORDERS
FIELD OF THE INVENTION
The present invention relates generally to a method for the prophylaxis and/or
treatment of
medical disorders, and in particular proliferative and/or inflammatory skin
disorders, and to
genetic molecules useful for same. The present invention is particularly
directed to genetic
molecules capable of modulating growth factor interaction with its receptor on
cells such as
epidermal keratinocytes to inhibit, reduce or otherwise decrease stimulation
of this layer of cells.
The present invention contemplates, in a particularly preferred embodiment, a
method for the
prophylaxis and/or treatment of psoriasis or neovascularization conditions
such as
neovascularization of the retina. The present invention is further directed to
the subject genetic
molecules in adjunctive therapy for epidermal hyperplasia, such as in
combination with UV
treatment, and to facilitate apoptosis of cancer cells and in particular
cancer cells comprising
keratinocytes.
BACKGROUND OF THE INVENTION
Bibliographic details of the publications numerically referred to in this
specification are
collected at the end of the description.
The reference to any prior art in this specification is not, and should not be
taken as, an
acknowledgment or any form of suggestion that that prior art forms part of the
common general
knowledge in Australia or any other country.
Psoriasis and other similar conditions are common and often distressing
proliferative and/or
inflammatory skin disorders affecting or having the potential to affect a
significant proportion
of the population. The condition arises from over proliferation of basal
keratinocytes in the
epidermal layer of the skin associated with inflammation in the underlying
dermis. Whilst a


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range of treatments have been developed, none is completely effective and free
of adverse side
effects. Although the underlying cause of psoriasis remains elusive, there is
some consensus
of opinion that the condition arises at least in part from over expression of
local growth factors
and their interaction with their receptors supporting keratinocyte
proliferation via keratinocyte
receptors which appear to be more abundant during psoriasis.
One important group of growth factors are the dermally-derived insulin-like
growth factors
(IGFs) which support keratinocyte proliferation. In particular, IGF-I and IGF-
II are ubiquitous
peptides each with potent mitogenic effects on a broad range of cells.
Molecules of the IGF type
are also known as "progression factors" promoting "competent" cells through
DNA synthesis.
The IGFs act through a common receptor known as the Type I or IGF-I receptor,
which is
tyrosine kinase linked. They are synthesised in mesenchymal tissues, including
the dermis, and
act on adjacent cells of mesodermal, endodermal or ectodermal origin. The
regulation of their
synthesis involves growth hormone (GH) in the liver, but is poorly defined in
most tissues [ 1 ].
Particular proteins, referred to as IGF binding proteins (IGFBPs), appear to
be involved in
autocrine/paracrine regulation of tissue IGF availability [2]. Six IGFBPs have
so far been
identified. The exact effects of the IGFBPs is not clear and observed effects
in vitro have been
inhibitory or stimulatory depending on the experimental method employed [3].
There is some
evidence, however, that certain IGFBPs are involved in targeting IGF-I to its
cell surface
receptor.
Skin, comprising epidermis and underlying dermis, has GH receptors on dermal
fibroblasts [4].
Fibroblasts synthesize IGF-I as well as IGFBPs-3, -4, -5 and -6 [5] which may
be involved in
targeting IGF-I to adjacent cells as well as to the overlaying epidermis. The
major epidermal
cell type, the keratinocyte, does not synthesize IGF-I, but possesses IGF-I
receptors and is
responsive to IGF-I [6].
It is apparent, therefore, that IGF-I and other growth promoting molecules,
are responsible for
or at least participate in a range of skin cell activities. In accordance with
the present invention,
the inventors have established that aberrations in the normal functioning of
these molecules or


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aberrations in their interaction with their receptors is an important factor
in a variety of medical
disorders such as proliferative and/or inflammatory skin disorders. It is
proposed, therefore, to
target these molecules or other molecules which facilitate their functioning
or interaction with
their receptors to thereby ameliorate the effects of aberrant activity during
or leading to skin
disease conditions and other medical conditions such as those involving
neovascularization.
Furthermore, these molecules may also be used to facilitate apoptosis of
target cells and may
be useful as adjunctive therapy for epidermal hyperplasia.
SUMMARY OF THE INVENTION
Nucleotide and amino acid sequences are referred to by a sequence identifier,
i.e. (<400>1),
(<400>2), etc. A sequence listing is provided after the claims.
Throughout this specification, unless the context requires otherwise, the word
"comprise", or
variations such as "comprises" or "comprising", will be understood to imply
the inclusion of a
stated element or integer or group of elements or integers but not the
exclusion of any other
element or integer or group of elements or integers.
Accordingly, one aspect of the present invention contemplates a method for
ameliorating the
effects of a medical disorder such as a proliferative and/or inflammatory skin
disorder in a
mammal, said method comprising contacting the proliferating and/or inflamed
skin or skin
capable of proliferation and/or inflammation or a cell otherwise involved in
the said medical
disorder with an effective amount of a nucleic acid molecule or chemical
analogue thereof
capable of inhibiting or otherwise reducing a growth factor mediated cell
proliferation and/or
inflammation and/or other medical disorder.
According to this preferred embodiment, there is provided a method for
ameliorating the
effects of a medical disorder such as a proliferative and/or inflammatory skin
disorder in a
mammal, said method comprising contacting the proliferating and/or inflamed
skin or skin
capable of proliferation and/or inflammation or a cell otherwise involved with
said medical
disorder with an effective amount of a nucleic acid molecule or chemical
analogue thereof


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capable of inhibiting or otherwise reducing IGF-I mediated cell proliferation
and/or
inflammation and/or other medical disorder.
According to this embodiment, there is provided a method for ameliorating the
effects of a
proliferative and/or inflammatory skin disorder such as psoriasis said method
comprising
contacting the proliferating and/or inflamed skin or skin capable of
proliferation and/or
inflammation with effective amounts of UV treatment and a nucleic acid
molecule or chemical
analogue thereof capable of inhibiting or otherwise reducing IGF-I mediated
cell proliferation
and/or inflammation.
According to this embodiment, there is provided in a particularly preferred
aspect a ribozyme
comprising a hybridising region and a catalytic region wherein the hybridising
region is capable
of hybridising to at least part of a target mRNA sequence transcribed from a
genomic gene
corresponding to <400>1 or <400>2 wherein said catalytic domain is capable of
cleaving said
target mRNA sequence to reduce or inhibit IGF-I mediated cell proliferation
and/or
inflammation and/or other medical disorders.
Yet another aspect of the present invention contemplates co-suppression to
reduce expression
or to inhibit translation of an endogenous gene encoding, for example, IGF-I,
its receptor, or
IGFBPs such as IGFBP-2 and/or -3. In co-suppression, a second copy of an
endogenous gene
or a substantially similar copy or analogue of an endogenous gene is
introduced into a cell
following topical administration. As with antisense molecules, nucleic acid
molecules defining
a ribozyme or nucleic acid molecules useful in co-suppression may first be
protected such as by
using a nonionic backbone.
Another aspect of the present invention contemplates a pharmaceutical
composition for topical
administration which comprises a nucleic acid molecule capable of inhibiting
or otherwise
reducing IGF-I mediated cell proliferation such as psoriasis and one or more
pharmaceutically
acceptable Garners and/or diluents.


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Yet another aspect of the present invention contemplates the use of a nucleic
acid molecule in
the manufacture of a medicament for the treatment of proliferative and/or
inflammatory skin
disorders or other medical disorders mediated by a growth factor.
Still a further aspect of the present invention contemplates an agent
comprising a nucleic acid
molecule as hereinbefore defined useful in the treatment of proliferative
and/or inflammatory
skin disorders, such as psoriasis or other medical disorder..
The present invention further contemplates the use of the genetic molecules
and in particular
the antisense molecules to inhibit the anti-apoptotic activity of IGF-I
receptor.


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BRIEF DESCRIPTION OF THE FIGURES
Figure 1 is a representation of the nucleotide sequence of IGFBP-2.
LOCUS HSIGFBP2 1433 by RNA PRI 31-JAN-1990


S DEFINITIONHuman mRNA for insulin-like growth factor binding
protein (IGFBP-2)


ACCESSION X16302


KEYWORDS insulin-like growth factor binding protein.


SOURCE human


ORGANISM Homo Sapiens


IO Eukaryota; Animalia; Metazoa; Chordata; Vertebrata;
Mammalia;


Theria; Eutheria; Primates; Haplorhini; Catarrhini;
Hominidae.


REFERENCE 1 (bases 1 to 1433)


AUTHORS Binkert,C., Landwehr,J., Mary,J.L., Schwander,J.
and Heinrich,G.


TITLE Cloning, sequence analysis and expression of a cDNA
encoding a


1S novel insulin-like growth factor binding protein
(IGFBP-2)


JOURNAL EMBO J. 8, 2497-2502 (1989)


STANDARD full automatic


COMMENT NCBI gi: 33009


FEATURES Location/Qualifiers


ZO source 1. .1433


/organism="Homo Sapiens"


/dev stage="fetal"


/tissue type="liver"


misc eature 1416. .1420
f


_ /note="pot. polyadenylation signal"
ZS


polyA site 1433


/note="polyadenylation site"


CDS 118. .1104


/note="precursor polypeptide; (AA -39 to 289); NCBI
gi:


3O 33010. "


/codon start=1


/translation="MLPRVGCPALPLPPPPLLPLLPLLLLLLGASGGGGGARAEVLFR


CPPCTPERLAACGPPPVAPPAAVAAVAGGARMPCAELVREPGCGCCSVCARLEGEACG


VYTPRCGQGLRCYPHPGSELPLQALVMGEGTCEKRRDAEYGASPEQVADNGDDHSEGG


3S LVENHVDSTMNMLGGGGSAGRKPLKSGMKELAVFREKVTEQHRQMGKGGKHHLGLEEP


KKLRPPPARTPCQQELDQVLERISTMRLPDERGPLEHLYSLHIPNCDKHGLYNLKQCK


MSLNGQRGECWCVNPNTGKLIQGAPTIRGDPECHLFYNEQQEACGVHTQRMQ"


(<400>21)


CDS 118. .234


4O /note="signal peptide; (AA -39 to -1); NCBI gi: 33011."


/codon start=1


/translation="MLPRVGCPALPLPPPPLLPLLPLLLLLLGASGGGGGARA"


(<400>22)


CDS 235. .1101


4S /note="mature IGFBP-2; (AA 1 to 289); NCBI gi: 33012."


/codon start=1


/translation="EVLFRCPPCTPERLAACGPPPVAPPAAVAAVAGGARMPCAELVR


EPGCGCCSVCARLEGEACGVYTPRCGQGLRCYPHPGSELPLQALVMGEGTCEKRRDAE


YGASPEQVADNGDDHSEGGLVENHVDSTMNMLGGGGSAGRKPLKSGMKELAVFREKVT


SO EQHRQMGKGGKHHLGLEEPKKLRPPPARTPCQQELDQVLERISTMRLPDERGPLEHLY


SLHIPNCDKHGLYNLKQCKMSLNGQRGECWCVNPNTGKLIQGAPTIRGDPECHLFYNE


QQEACGVHTQRMQ" (<400>23)


BASE COUNT 239 a 466 c 501 g 227 t


ORIGIN
SS


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_ '7 _
HSIGFBP2 Length: 1433 May 11, 1994 10:06 Type: N Check: 6232 ..
Figure 2 is a representation of the nucleotide sequence of IGFBP-3.
S
LOCUS HUMGFIBPA 2474 by ss-mRNA PRI 15-JUN-1990
DEFINITION Human growth hormone-dependent insulin-like growth factor-binding
protein mRNA, complete cds.
ACCESSION M31159
1~ KEYWORDS insulin-like growth factor binding protein.
SOURCE Human plasma, cDNA to mRNA, clone BP-53.
ORGANISM Homo sapiens
Eukaryota; Animalia; Chordata; Vertebrata; Mammalia; Theria;
Eutheria; Primates; Haplorhini; Catarrhini; Hominidae.
1S REFERENCE 1 (bases 1 to 2474)
AUTHORS Wood,W.I., Cachianes,G., Henzel,W.J., Winslow,G.A., Spencer,S.A.,
Hellmiss,R., Martin,J.L. and Baxter,R.C.
TITLE Cloning and expression of the growth hormone-dependent insulin-like
growth factor-binding protein
JOURNAL Mol. Endocrinol. 2, 1176-1185 (1988)
STANDARD full automatic
COMMENT NCBI gi: 183115
FEATURES Location/Qualifiers
mRNA <1. .2474
ZS /note="GFIBP mRNA"
CDS 110. .985
/gene="IGFBP1"
/note="insulin-like growth factor-binding protein; NCBI
gi: 183116."
/codon start=1
/translation="MQRARPTLWAAALTLLVLLRGPPVARAGASSGGLGPWRCEPCD
ARALAQCAPPPAVCAELVREPGCGCCLTCALSEGQPCGIYTERCGSGLRCQPSPDEAR
PLQALLDGRGLCVNASAVSRLRAYLLPAPPAPGNASESEEDRSAGSVESPSVSSTHRV
SDPKFHPLHSKIIIIKKGHAKDSQRYKVDYESQSTDTQNFSSESKRETEYGPCRREME
3S DTLNHLKFLNVLSPRGVHIPNCDKKGFYKKKQCRPSKGRKRGFCWCVDKYGQPLPGYT
TKGKEDVHCYSMQSK" (<400>24>)
source 1. .2474
/organism="Homo sapiens"
BASE COUNT 597 a 646 c 651 g 580 t
4O ORIGIN
HUMGFIBPA Length: 2474 May 11, 1994 10:00 Type: N Check: 9946 ..
4S Figure 3 is a representation of the nucleotide sequence of IGF-1-receptor.
LOCUS HSIGFIRR 4989 by RNA PRI 28-MAR-1991
DEFINITION Human mRNA for insulin-like growth factor I receptor
ACCESSION X04434 M24599
$0 KEYWORDS glycoprotein; insulin receptor;
insulin-like growth factor I receptor; membrane glycoprotein;
receptor; tyrosine kinase.
SOURCE human


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_g_


ORGANISM
Homo
sapiens


Eukaryota;
Animalia;
Metazoa;
Chordata;
Vertebrata;
Mammalia;


Theria; Eutheria; Primates; Haplorhini; Catarrhini;
Hominidae.


REFERENCE1 (bases
1 to
4989)


AUTHORSUllrich,A., .,
Gray,
A.,
Tam,A.W.,
Yang-Feng,T.,
Tsubokawa,M


Collins,C.,
Henzel,W.,
Bon,T.L.,
Kathuria,S.,
Chen,E.,
Jakobs,S.,


Francke,U.,
Ramachandran,J.
and
Fujita-Yamaguchi,Y.


TITLE Insulin-like growth factor I receptor primarycomparison
structure:


with
insulin
receptor
suggests
structural
dererminants
that
define


1~ functional
specificity


JOURNALEMBO 5, 2503-2512 (1986)
J.


STANDARDfull
automatic


COMMENTNCBI 33058
gi:


FEATURES Location/Qualifiers


IS source 1. .4989


/organism="Homo sapiens"


/tissue type="placenta"


/clone lib="(lamda)gtl0"


/clone="(lambda)IGF-1-R.85, (lambda)IGF-1-R.76"


sig~eptide 32. .121


mat 122. .4132
peptide


/note="IGF-I receptor"


misc_ feature 122. .2251


/note="alpha-subunit (AA 1 - 710)"


25 misc_ feature 182. .190


/note="pot.N-linked glycosylation 23)"
site (AA 21 -


misc_ feature 335. .343


/note="pot.N-linked glycostlation 74)"
site (AA 72 -


misc_ feature 434. .442


/note="pot.N-linked glycostlation - 107)"
site (AA 105


misc_ feature 761. .769


/note="pot.N-linked glycostlation - 216)"
site (AA 214


misc_ feature 971. .979


/note="pot.N-linked glycostlation - 286)"
site (AA 284


35 misc_ feature 1280. .1288


/note="pot.N-linked glycostlation - 389)"
site (AA 387


misc_ feature 1343. .1351


/note="pot.N-linked glycosylation - 410)"
site (AA 408


misc feature 1631. .1639


_ /note="pot.N-linked glycostlation - 506)"
site (AA 504


misc_ feature 1850. .1858


/note="pot.N-linked glycosylation - 579)"
site (AA 577


misc_ feature 1895. .1903


/note="pot.N-linked glycosylation - 594)"
site (AA 592


45 misc_ feature 1949. .1957


/note="pot.N-linked glycosylation - 612)"
site (AA 610


misc_ feature 2240. .2251


/note="putative proreceptor processing707 -
site (AA


710)"


$0 misc_ feature 2252. .4132


/note="beta-subunit (AA 711 - 1337)"


misc_ feature 2270. .2278


/note="pot.N-linked glycosylation - 719]"
site (AA 717


misc_ feature 2297. .2305


$S /note="pot.N-linked glycosylation - 728)"
site (AA 726


misc_ feature 2321. .2329


/note="pot.N-linked glycosylation 736)"
site (AA 734 -




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misc_ feature 2729. .2737


/note="pot.N-linked glycosylation(AA 870 - 872)"
site


misc_ feature 2768. .2776


/note="pot.N-linked glycosylation(AA 883 - 885)"
site


misc_ feature 2837. .2908


/note="transmembrane region 929)"
(AA 906 -


misc_ feature 2918. .2926


/note="pot.N-linked glycosylation(AA 933 - 935)"
site


misc feature 3047. .3049


10_ /note="pot. ATP binding site
(AA 976)"


misc_ feature 3053. .3055


/note="pot. ATP binding site
(AA 978)"


misc_ feature 3062. .3064


/note="pot. ATP binding site
(AA 981)"


15misc_ feature 3128. .3130


/note="pot. ATP binding site
(AA 1003)"


CDS 32. .4132


/product="IGF-I receptor"


/note="50 stops when translation
attempted, frame 1, code


20 0"


BASE 1216 a 1371 c 1320 g 1082 t
COUNT


ORIGIN


HSIGFIRR Length: 4989 May 11, 1994 12:10 Type: N Check: 133 ..
Figure 4A is a photographic representation of a Western ligand blot of HaCaT
conditioned
medium showing IGFBP-3 secreted in 24 hours after 7 day treatment with
phosphorothioate
oligonucleotides (BP3AS2, BP3AS3 and BP3S) at O.S~.M and S~,M;
* no oligonucleotide added.
Figure 4B is a graphical representation of a scanning imaging desitometry of
Western ligand
blot (Figure 4A), showing relative band intensities of IGFBP-3 and the 24kDa
IGFBP-4 after
treatment with phosphorothioate oligonucleotides;
* no oligonucleotide added.
Figure 5A is a photographic representation of a Western ligand blot of HaCaT
conditioned
medium showing IGFBP-3 secreted in 24 hours after 7 day treatment with
phosophorothioate
oligonucleotide BP3AS2 at O.S~,M compared with several control
oligonucleotides at O.S~.M.
(a) oligonucleotide BP3AS2NS; (b) oligonucleotide BP3AS4; (c) oligonucleotide
BP3AS4NS; and (untreated), no oligonucleotide added.


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Figure 5B is a graphical representation of a scanning imaging densitometry of
Western ligand
blot (Figure SA), showing relative band intensities of IGFBP-3 after treatment
with
phosphorothioate oligonucleotides as in Figure SA, showing IGFBP-3 band
intensities
expressed as a percentage of the average band intensity from conditioned
medium of cells not
treated with oligonucleotide.
Figure 6 is a graphical representation showing inhibition of IGF-I binding by
antisense
oligonucleotides to IGF-I receptor. IGFR.AS: antisense; IGFR.S: sense.
Figure 7 is a graphical representation showing inhibition of IGFBP-3
production in culture
medium following initial treatment with antisense oligonucleotides once daily
over a 2 day
period.
Figure 8 is a graphical representation showing optimization of IGFBP-3
antisense
oligonucleotide concentration as determined by relative IGFBP-3 concentration
in culture
medium.
Figure 9 is a diagramatic representation of a map of IGF-1 Receptor mRNA and
position of
target ODNs.
Figure 10 is a photographical representation showing Lipid-mediated uptake of
oligonucleotide in keratinocytes. HaCaT keratinocytes were incubated for 24
hours in medium
(DMEM plus 10% v/v FCS) containing fluorescently labelled ODN (R451, 30 nM)
and
cytofectin GSV (2 ~g/ml). The cells were then transferred to ODN-free medium
and
fluorescence microscopy (a) and phase contrast (b) images of the cells were
obtained.
Figure 11 is a graphical representation of uptake (A) and toxicity (B) of
ODN/lipid complexes
in keratinocytes. Confluence HaCaT keratinocytes were incubated in DMEM
containing
fluoresently labelled ODN (R451) plus liposome over 120 hours, viewed using
fluorescene
microscopy and trypan blue stained and counted.


CA 02376284 2001-12-05
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Figure 12 is a graphical representation of an IGF-1 Receptor mRNA in ODN
treated (30nM)
HaCaT cells (2~cg/ml GSV). HaCaT keratinocytes were treated for 96 hours with
C-5
propynyl, dU, dC ODNs complexed with cytofectin GSV. Cells were treated with
ODNs
complementary to the human IGF-I receptor mRNA (27, 32, 74 and 78), 2
randomised
sequence ODNs (R451) and R766), liposome alone (GSV) or were left untreated
(UT). Total
RNA was isolated then analysed for IGF-I receptor mRNA and GAPDH mRNA levels
by
RNase Protection and Phosphorlmager quantitiation.
(A) Electrophoretic analysis of IGF-I receptor and GAPDH mRNA fragments after
RNase
Protection. Molecular weight markers are shown on the right hand side. Full
length probe is
shown on the left hand side (G-probe and I-probe). GAPDH protected fragments
(G) are seen
at 316 bases and IGF-I receptor protected fragments (I) are seen at 276 bases.
(B) Relative level of IGF-I receptor mRNA following each treatment is shown.
Figure 13 is a graphical representation of an IGF-1 receptor mRNA in ODN
treated (30nM)
HaCaT cells (2,ug/ml GSV). Summary of IGF-I receptor ODN screening data. HaCaT
keratinocytes were treated for 96 hours with C-5 propynyl, dU, dC ODNs
complexed with
cytofectin GSV. Total RNA was isolated then analysed for IGF-I receptor mRNA
and
GAPDH mRNA levels by RNase protection and phosphorImager quantitiation.
Relative level
of IGF-I receptor mRNA is shown after treatment with ODNs complementary to the
human
IGF-I receptor mRNA, 4 randomised sequence ODNs and liposome alone. (26-86=IGF-
I
receptor ODNs; R1, R4, R7 and R9 = randomised ODNs (R1=8121, R4=8451, R7=8766,
R9=R961); GSV=liposome alone; UT=untreated). *indicates a significant
difference in
relative IGF-I receptor mRNA from GSV treated cells (n=4-10, p < 0.05).
Figure 14 is a graphical representation of the effect of antisense
oligonucleotides on IGF-1
receptor levels on the surface of keratinocytes. HaCaT cells were grown to
confluence in 24-
well plates in DMEM containing 10% v/v FCS. Oligodeoxynucleotide (ODN) and
Cytofectin
GSV (GSV, Glen Research) were mixed together in serum-free DMEM, incubated at
room


CA 02376284 2001-12-05
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-12-
temperature for 10 minutes before being diluted ten-fold in medium and placed
on the cells.
Cells were incubated for 72 hours with 30 nM random sequence or antisense ODN
and 2
~cg/ml GSV or with GSV alone in DMEM containing 10% v/v FCS with solutions
replaced
every 24 hours. This was followed by incubation with ODN/GSV in serum-free
DMEM for
48 hours. All incubations were performed at 37°C. Wells were washed
twice with 1 ml cold
PBS. Serum-free DMEM containing 10-'°M 'z5I-IGF-I was added with or
without the IGF-I
analogue, des (1-3) IGF-I, at 10-'°M to 10-'M. Cells were incubated at
4°C for 17 hours with
gentle shaking then washed three times with 1 ml cold PBS and lysed in 250 ,u1
O.SM
NaOH/0.1 % v/v Triton X-100 at room temperature for 4 hours. Specific binding
of the
solubilised cell extract was measured using a y counter.
Figure 15 is a graphical representation of the effect of antisense
oligonucleotides on IGF-1
receptor levels on the surface of keratinocytes.
Figure 16 is a photographical representation of H & E stained sections of (A)
psoriatic skin
biopsy prior to grafting and (B) 49 day old psoriatic skin graft using skin
from the same
donor.
Figure 17 is a photographical representation of uptake of oligonucleotide
after intradermal
injection into psoriatic skin graft on a nude mouse. Psoriatic skin graft was
intradermally
injected with ODN (R451, 50 ,u1, 10 ~cM). The graft was removed and sectioned
after 24
hours, then viewed using confocal microscopy.
Figure 18(a) is a photographical representation of Pregraft, Donor JH, Donor
JH, PBS
treated, SO~cI, Donor JH, #50 treated, SO,uI, lO,uM.
Figure 18(b) is a photographical representation of Donor LB, pregraft, Donor
LB, PBS
treated (SO~cI), Donor LB, #74 treated (SO,uI, lO,uM).


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Figure 18(c) is a photographical representation of Donor PW, pregraft, Donor
PW, R451
treated (50,u1, lO~cM), Donor LB, #74 treated (50,u1, lO,uM).
Figure 18(d) is a photographical representation of Donor GM, pregraft, Donor
GB, 8451
treated (50,u1, lO,uM), Donor GM, #27 treated (50,u1, lO,uM).
Figure 19(a) is a photographical representation showing Donor JH pregraft,
Donor JH PBS
treated 50,u1, Donor JH #50 treated 50,u1, lO,uM.
Figure 19(b) is a photographical representation Donor LB pregraft, Donor LB
PBS treated
50,u1, Donor LB #74 treated 50,u1, lO,uM.
Figure 19(c) is a photographical representational showing Donor PW pregraft,
Donor PW
r451 treated 501, lO,uM, Donor PW #74 treated 50~c1, IO~cM.
Figure 19(d) is a photographical representation showing Donor GM pregraft,
Donor GM
8451 treated 501, 10~M, Donor #27 treated 50,u1, 10~M.
Figure 20 is a graphical representation showing suppression of psoriasis after
treatment with
oligonucleotide (quantification). Oligonucleotide (50 ,u1, lO,uM) was injected
every two days
for 20 days, as were control treatments. Skin thickness was measured by
removing the skin
and using computer software (MCID analysis) to measure the exact thickness of
each graft.
N=3-4 for each treatment. *indicates a significant difference from the
pregraft value
(ANOVA, P < 0.05)
Figure 21 is a photographic representation of ahKi-67 imunobiological binding.
Figure 22 is a photographical representation showing penetration of
oligonucleotide into
human skin after topical treatment. Fluorescently labelled oligonucleotide (10
~cM R451) was


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applied topically after formulation with cytofectin GSV (10 ,ug/ml) and viewed
using confocal
microscopy.
Figure 23 is a photographical representation showing penetration of
oligonucleotide into
human skin after application of topical gel formation. Fluorescently labelled
oligonucleotide
(10 ,uM R451) was applied topically after complexing with cytofectin GSV (10
,ug/ml) and
formulation into 3 % methylcellulose gel. Image was obtained using confocal
microscopy.
Figure 24 is a graphical representation showing IGFBP-3 mRNA.
Figure 25(a) is a graphical representation showing IGFBP-3 mRNA in AON treated
( 100nM)
HaCaT cells (2,ug/ml GSV).
Figure 25(b) is a graphical representation showing IGFBP-3 mRNA levels of AON
treated
(100nm) HaCaT cells (2~cg/ml GSV).
Figure 25(c) is a graphical representation showing IGFBP-3 mRNA in AON treated
(30nM)
HaCaT cells (2~cg/ml GSV).
Figure 25(d) is a graphical representation showing IGFBP-3 mRNA in AON treated
(30nM)
HaCaT cells (2,ug/ml GSV).
Figure 26(a) is a graphical representation showing IGFBP-3 mRNA in ODN treated
(30nM)
HaCaT cells (2,ug/ml). HaCaT keratinocytes were treated for 51 hours with C-5
propynl, dU,
dC ODNs complexed with cytofectin GSV. Total RNA was isolated then analysed
for IGFBP-
3 mRNA and GAPDH mRNA levels by Northern analysis and phosphorimager
quantitation.
Relative level of IGFBP-3 mRNA is shown after treatment with ODNs
complementary to the
human IGFBP-3 mRNA, 4 randomised sequence ODNs and lipsome alone. (1-24=IGFBP-
3
ODNs; R1, R4, R7 and R9=randomised ODNs (R1=8121, R4=8451, R7=8766, R9
R961); GS=liposome alone; UT=untreated). *indicates a significant different in
relative


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IGFBP-3 mRNA from GSV treated cells (n- 5-8, p < 0.01), **indicates a
significant difference
in relative IGFBP-3 mRNA from GSV treated cells (n= 5-8, p < 0.05).
Figure 26(b) is a graphical representation showing IGFBP-3 mRNA in ODN treated
(100nM)
HaCaT cells (2,ug/ml GSV). HaCaT keratinocytes were treated for 51 hours with
C-5
propynl, dU, dC ODNs complexed with cytofectin GSV. Total RNA was isolated
then
analysed for IGFBP-3 mRNA and GAPDH mRNA levels by Northern analysis and
phosphorimager quantitation. Relative level of IGFBP-3 mRNA is shown after
treatment with
ODNs complementary to the human IGFBP-3 mRNA, 4 randomised sequence ODNs and
liposome alone. (1-24=IGFBP-3 ODNs; R1, R4, R7 and R9 = randomised ODNs (Rl-
R121,
R4=8451, R7=8766, R9-R961), GS=lipsome alone; UT=untreated). *indicates a
significant difference in relative IGFBP-3 mRNA from GSV treated cells (n- 6-
8, p < 0.01).
Figure 27 is a representation showing a reduction in IGF-I receptor mRNA in
HaCaT cells
following treatment with antisense oligonucleotides. Confluent HaCaT cells
were treated
every 24 h for 4 days with 2 ,ug/ml GSV lipid alone (GSV) or complexed with 30
nM IGF-I
receptor specific oligonucleotides (#26 to #86) or random sequence
oligonucleotides (R121,
8451 and R766). Total RNA was isolated and analysed for IGF-I receptor and
GAPDH
mRNA by RNase protection assay. (a). Representative RNase protection assay gel
showing
IGF-I receptor (IGFR) and GAPDH mRNA in untreated or treated HaCaT cells. In
this
example, a reduction in IGFR band intensity relative to GAPDH can be seen with
AON #27
and #78, but not with #32, #74 or the controls (R4, R7, random
oligonucleotides 8451 and
8766, respectively; G, GSV lipid; UT, untreated).
(b) Densitometric quantitation of IGF-I receptor mRNA (normalised to GAPDH
mRNA) in
HaCaT cells following treatment with IGF-I receptor specific oligonucleotides
(solid black),
random sequence oligonucleotides (horizontal striped bar) or GSV alone (shaded
bar)
compared to untreated cells (UT, vertical striped bar). Each oligonucleotide
was assayed in
duplicate in at least two separate experiments.


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Results are presented as mean t SEM. A one-way ANOVA followed by Tukey's (1)
test
was performed; 1 indicates a significant difference between cells treated with
IGF-I receptor
specific AONs and all of the control treatments (p < 0.05). n=4 except for #27
and #32
(n=6), #28 and #68 (n=3), 8766 (n=9), and 8451, GSV and untreated (n=10).
S
Figure 28 is a representation showing a reduction in total cellular IGF-I
receptor protein
following antisense oligonucleotide treatment. Confluent HaCaT cells were
treated every 24
h for 4 days with 2 ,ug/ml GSV lipid alone (GSV) or complexed with 30 nM IGF-I
receptor
specific AONs (#27, #50 and #64) or the random sequence oligonucleotide, 8451.
Total
cellular protein was isolated and analysed for IGF-I receptor by SDS PAGE
followed by
western blotting with an antibody specific for the human IGF-I receptor. (a)
Duplicate treated
cellular extracts showing the IGF-I receptor at the predicted size of 110 kD
(b) Densitometric quantitation of IGF-I receptor protein. Results are
presented as mean ~
SEM of four different experiments each performed in duplicate. A one-way ANOVA
followed
by a Dunnett's test was performed; * indicates a significant difference from
GSV treated cells
(p<0.01). GfV, GSV lipid alone; UT, untreated; 8451, random sequence
oligonucleotide;
64, S0, 27, IGF-I receptor-specific AONs.
Figure 29 is a representation showing a reduction in IGF-I receptor numbers on
the
keratinocyte cell surface after antisense oligonucleotide treatment. HaCaT
cells were
transfected with IGF-I receptor specific AONs #27 (-1-), #50 (-x-), #64 (---~--
-), a
random sequence oligonucleotide 8451 (-o-), or treated with GSV lipid alone (--
~--)
every 24 h for four days (untreated cells, --~--). Competition binding assays
using l2sl_IGF-I
and the receptor-specific analogue, des(1-3)IGF-I, were performed (inset);
plotted values are
means t standard error. The mean values were then subjected to Scatchard
analysis.
Figure 30 is a representation showing a reduction in keratinocyte cell number
following
antisense oligonucleotide treatment. HaCaT cells, initially at 40 %
confluence, were
transfected with the IGF-I receptor specific AON #64, control sequences 8451
and 6416, or


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treated with GSV lipid alone every 24 h for 2 days (UT, untreated cells). Cell
number was
measured in the culture wells using a dye binding assay (Experimental
protocol). Results are
presented as mean ~ SD. A one-way ANOVA was performed, followed by a Tukey's
multiple comparison test. 1 indicates a significant difference between cells
treated with
AON #64 and all of the control treatments (p < 0.001).
Figure 31 is a representation showing a reversal of epidermal hyperplasia in
psoriatic human
skin grafts on nude mice following intradermal injection with antisense
oligonucleotides
Grafted psoriasis lesions were injected with IGF-I receptor specific AONs, a
random sequence
oligonucleotide in PBS, or with PBS alone, every 2 days for 20 days, then
analysed
histologically. (a) Donor A graft treated with AON #50 showing epidermal
thinning compared
with pregraft and control (PBS) treated graft, and Donor B graft treated with
AON #27
showing epidermal thinning compared with pregraft and control (R451) treated
graft. E,
epidermis; Scale bar, 400 mm; all pictures are at the same magnification. (b)
Mean epidermal
cross-sectional area over the full width of grafts was determined by digital
image analysis.
Results are presented as mean ~ SEM. Shaded bars, control treatments: 8451,
random
oligonucleotide sequence; solid bars, treatments with oligonucleotides that
inhibited IGF-I
receptor expression in vitro. * indicates a significant difference from the
vehicle treated graft
(p < 0.01, n=5-7), + + indicates a significant difference from the random
sequence (R451)
treated graft (p<0.01, n=5-7). (c) Parakeratosis (arrow) was absent in grafts
treated with
IGF-I receptor AONs (AON #50) but persisted in pregraft and control (PBS)
treated graft.
Scale bar, 100 mm.
Figure 32 is a representation showing a reversal of epidermal hyperplasia
correlates with
reduced IGF-I receptor mRNA in grafted psoriasis lesions treated with
antisense
oligonucleotides (a) A psoriasis lesion prior to grafting, and after grafting
and treatment with
IGF-I receptor specific oligonucleotide #27 (AON #27) or random sequence
(R451) was
immunostained with antibodies to Ki67 to identify proliferating cells.
Proliferating cells are
indicated by a dark brown nucleus (arrows). Scale bar, 250 mm; all pictures
are at the same


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-18-
magnification. (b) The same lesion prior to grafting and after oligonucleotide
treatment as in
(a) was subjected to in situ hybridisation with a 35S-labeled cRNA probe
complementary to
the human IGF-I receptor mRNA. The presence of IGF-I receptor mRNA is
indicated by
silver grains (tiny black speckles), which are almost eliminated in the
epidermis of the lesion
treated with the IGF-I receptor-specific oligonucleotide #27 (AON #27). Arrows
indicate the
basal layer of the epidermis with dermis underneath. Scale bar, 50 ,um.
Figure 33 is a representation showing a reduction in IGF-I receptor mRNA in
HaCaT
keratinocytes following treatment with oligonucleotides. HaCaT cell monolayers
grown to
90 % confluence in DMEM contianing 10 % v/v fetal calf serum were treated with
24 h for
two days with 2 ,ug/ml GSV lipid alone (GSV) or complexed with 30 nM
oligonucleotide.
Total RNA was isolated and analysed for IGF-I receptor and GAPDH mRNA using a
commercially availble ribonuclease protection assay kit (RPAII, Ambicon Inc,
Austin, Texas).
Band intensity was quantified using ImageQuant software (Molecular Dynamics,
Sunnyvale,
California).
Figure 34 is a representation showing a reduction in IGF-I receptor protein in
HaCaT
keratinocytes following treatment with oligonucleotides. HaCaT cell monolayers
grown to
90 % confluence in DMEM containing 10 % v/v fetal calf serum were treated
every 24 h for
four days with 2 ~g/ml GSV lipid alone (GSV) or complexed with 30 nM
oligonucleotide.
Cells were lyased in a buffer containing 50 mM HEPES, 150 mM NaCI, 10 % v/v
gycerol,
1 % v/v Triton X-100 and 100 ~g/ml aprotinin on ice for 30 mins, then 30 ,ug
of lysate was
loaded onto a denaturing 7 % w/v polyacrylamide gel followed by transfer onto
an Immobilon-
P membrane (Millipore, Bedford, Massachusetts). Membranes were incubated with
the anti-
IGF-I receptor antibody C20 (Sanra Cruz Biotechnology Inc., Santa Cruz,
California, 25
ng/ml in 150 mM NaCI, 10 mM Tris-HCI, pH 7.4, 0.1 % v/v Tween 20) for 1 h at
room
temperature and developed using the Vistra ECF western blotting kit (Amersham,
Buckinghamshire, England). Band intensity was quantified using ImageQuant
software
(Molecular Dynamics, Sunnyvale, California).


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Figure 35 is a representation showing a reduction in HaCaT keratinocyte cell
number
following treatment with oligonucleotides. HaCaT cell monolayers grown to 40%
confluence
in DMEM containing 10 % fetal calf serum were treated every 24 h for three
days with 2
,ug/ml GSV lipid alone (GSV) or complexed with 15 nM oligonucleotide. Cell
number was
measured every 24 h using the amido black dye binding assay [32].


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DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
The present invention is predicated in part on the use of molecules and in
particular genetic
molecules and more particularly antisense molecules to down-regulate a growth
factor, its
receptor and/or growth factor expression facilitating sequences.
Accordingly, one aspect of the present invention contemplates a method for
ameliorating the
effects of a medical disorder such as a proliferative and/or inflammatory skin
disorder in a
mammal, said method comprising contacting the proliferating and/or inflamed
skin or skin
capable of proliferation and/or inflammation or a cell otherwise involved in
the said medical
disorder with an effective amount of a nucleic acid molecule or chemical
analogue thereof
capable of inhibiting or otherwise reducing a growth factor mediated cell
proliferation and/or
inflammation and/or other medical disorder.
Growth factor mediated cell proliferation and inflammation are also referred
to as epidermal
hyperplasias and these and other medical disorders may be mediated by any
number of
molecules such as but not limited to IGF-I, keratinocyte growth factor (KGF),
transforming
growth factor-a (TGFa), tumour necrosis factor-a (TNFa), interleukin-1, -4, -6
and 8 (IL-1,
IL-4, IL-6 and IL-8, respectively), basic fibroblast growth factor (bFGF) or a
combination
of one or more of the above. The present invention is particularly described
and exemplified
with reference to IGF-I and its receptor (IGF-I receptor) and to IGF-I
facilitating molecules,
IGFBPs, since targeting these molecules according to the methods contemplated
herein
provides the best results to date. This is done, however, with the
understanding that the
present invention extends to any growth factor or cytokine-like molecule, a
receptor thereof
or a facilitating molecule like the IGFBPs involved in skin cell proliferation
such as those
molecules contemplated above and/or their receptors and/or facilitating
molecules therefor.
According to this preferred embodiment, there is provided a method for
ameliorating the
effects of a medical disorder such as a proliferative and/or inflammatory skin
disorder in a
mammal, said method comprising contacting the proliferating and/or inflamed
skin or skin


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capable of proliferation and/or inflammation or a cell otherwise involved with
said medical
disorder with an effective amount of a nucleic acid molecule or chemical
analogue thereof
capable of inhibiting or otherwise reducing IGF-I mediated cell proliferation
and/or
inflammation and/or other medical disorder.
The present invention is particularly described by psoriasis as the
proliferative skin disorder.
However, the subject invention extends to a range of proliferative and/or
inflammatory skin
disorders or epidermal hyperplasias such as but not limited to psoriasis,
ichthyosis, pityriasis
rubra pilaris ("PRP"), seborrhoea, keloids, keratoses, neoplasias and
scleroderma, warts,
benign growths and cancers of the skin. The present invention extends to a
range of other
disorders such as neovascularization conditions such as but not limited to
hyperneovasularization such as neovascularization of the retina, lining of the
brain, skin,
hyperproliferation of the inside of blood vessels, kidney disease,
atherosclerotic disease,
hyperplasias of the gut epithelium or growth factor mediated malignancies such
as IGF1-
mediated malignancies.
Furthermore, down-regulation of IGF-I receptor is useful as adjunctive therapy
for epidermal
hyperplasia. In accordance with this aspect of the present invention it is
known that IGF-I
receptor elicits separate intracellular signals which prevent apoptosis [19].
In keratinocytes,
IGF-I receptor activation has been shown to protect UV-irradiated cells from
apoptosis [20].
In another cell type, a number of IGF-I receptors expressed by the cells
correlated with
tumorigenicity and apoptotic resistance [21]. Consequently, in accordance with
the present
invention, by inactivating IGF-I receptor on cells such as epidermal
keratinocytes will achieve
three important outcomes:
(r) Acute epidermal hyperplasia following UV has been suggested to increase
the risk of
keratinocyte carcinogenic transformation [22]. By reducing IGF-I receptor
expression
in the epidermis, the incidence of epidermal hyperplasia following UV exposure
is
likely to be reduced leading to an overall acceleration in normalization of
the lesion
and reduced carcinogenic risk.


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(ii) Inhibition of anti-apoptotic action of IGF-I receptor will enhance the
reversal of
epidermal thickening and accelerate normalization of differentiation. Topical
or
injected IGF-I receptor antisense as adjunctive treatment will increase
apoptosis in the
epidermal layer thereby enhancing the reduction in acanthosis observed in UV
treatments.
(iii) Survival of keratinocytes, ie. those which evade apoptosis is likely to
occur when cells
have damaged DNA. Such mutations may be in the tumor suppressor region.
Consequently, the use of antisense therapy will result in less frequent
selection of
mutated keratinocytes and therefore reduced incidence of basal cell carcinomas
and
squamous.
According to this embodiment, there is provided a method for ameliorating the
effects of a
proliferative and/or inflammatory skin disorder such as psoriasis said method
comprising
1 S contacting the proliferating and/or inflamed skin or skin capable of
proliferation and/or
inflammation with effective amounts of UV treatment and a nucleic acid
molecule or chemical
analogue thereof capable of inhibiting or otherwise reducing IGF-I mediated
cell proliferation
and/or inflammation.
The UV treatment and nucleic acid molecule or its chemical analogue may be
administered
in any order or may be done simultaneously. This method is particularly useful
in treating
psoriasis by combination of UV and antisense therapy. Preferably the antisense
therapy is
directed to the IGF-I receptor.
In a preferred embodiment, the present invention is directed to a method for
ameliorating the
effects of psoriasis or other medical disorder, said method comprising
contacting proliferating
skin or skin capable of proliferation or cells associated with said disorder
with an effective
amount of a nucleic acid molecule or chemical analogue thereof capable of
inhibiting or
otherwise reducing IGF-I mediated cell proliferation or ameliorating the
medical disorder.


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The present invention extends to any mammal such as but not limited to humans,
livestock
animals (e.g. horses, sheep, cows, goats, pigs, donkeys), laboratory test
animals (e.g. rabbits,
mice, guinea pigs), companion animals (e.g. cats, dogs) and captive wild
animals. However,
the instant invention is particularly directed to proliferative and/or
inflammatory skin
disorders such as psoriasis in humans as well as medical disorders
contemplated above.
The aspects of the subject invention instantly contemplated are particularly
directed to the
topical application of one or more suitable nucleic molecules capable of
inhibiting, reducing
or otherwise interfering with IGF-mediated cell proliferation and/or
inflammation. More
particularly, the nucleic acid molecule targets IGF-I interaction with its
receptor.
Conveniently, therefore, the nucleic acid molecule is an antagonist of IGF-I
interaction with
its receptor. Most conveniently, the nucleic acid molecule antagonist is an
antisense molecule
to the IGF-I receptor, to IGF-I itself or to a molecule capable of
facilitating IGF-I interaction
with its receptor such as but not limited to an IGFBP.
Insofar as the invention relates to IGFBPs, the preferred molecules are IGFBP-
2, -3, -4,
-5 and -6. The most preferred molecules are IGFBP-2 and IGFBP-3.
The nucleotide sequences of IGFBP-2 and IGFBP-3 are set forth in Figures 1 ( <
400 > 1) and
2 ( < 400 > 2), respectively. According to a particularly preferred aspect of
the present
invention, there is provided a nucleic acid molecule comprising at least about
ten nucleotides
capable of hybridising to, forming a heteroduplex or otherwise interacting
with an mRNA
molecule directed from a gene corresponding to a genomic form of < 400 > 1
and/or
< 400 > 2 and which thereby reduces or inhibits translation of said mRNA
molecule.
Preferably, the nucleic acid molecule is at least about 15 nucleotides in
length and more
preferably at least about 20-25 nucleotides in length. However, the instant
invention extends
to any length nucleic acid molecule including a molecule of 100-200
nucleotides in length to
correspond to the full length of or near full length of the subject genes.


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The nucleotide sequence of the antisense molecules may correspond exactly to a
region or
portion of < 400 > 1 or < 400 > 2 or may differ by one or more nucleotide
substitutions,
deletions and/or additions. It is a requirement, however, that the nucleic
acid molecule
interact with an mRNA molecule to thereby reduce its translation into active
protein.
Examples of potential antisense molecules for IGFBP-2 and IGFBP-3 are those
capable of
interacting with sequences selected from the lists in Examples 6 and 7,
respectively.
The nucleic acid molecules in the form of an antisense molecule may be linear
or covalently
closed circular and single stranded or partially double stranded. A double
stranded molecule
may form a triplex with target mRNA or a target gene. The molecule may also be
protected
from, for example, nucleases, by any number of means such as using a nonionic
backbone or
a phosphorothioate linkage. A convenient nonionic backbone contemplated herein
is
ethylphosphotriester linkage or a 2'-O-methylribosyl derivative. A
particularly useful
modification modifies the DNA backbone by introducing phosphorothioate
internucleotide
linkages. Alternatively or in addition to the pyrimidine bases are modified by
inclusion of a
C-5 propyne substitution which modification is proposed to enhance duplex
stability [23]. The
present invention extends to any chemical modification to the bases and/or RNA
or DNA
backbone. Reference to a "chemical analogue" of a nucleic acid molecule
includes reference
to a modified base, nucleotide, nucleoside or phosphate backbone.
Examples of suitable oligonucleotide analogues are conveniently described in
Ts' O et al [7] .
Further suitable examples of oligonucleotide analogues and chemical
modifications are
described in references 25 to 31.
Alternatively, the antisense molecules of the present invention may target the
IGF-I gene itself
or its receptor or a multivalent antisense molecule may be constructed or
separate molecules
administered which target at least two or an IGFBP, IGF-I and/or IGF-I-
receptor. Examples
of suitable antisense molecules capable of targetting the IGF-I receptor are
those capable of


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interacting with sequences selected from the list in Example 8. One
particularly useful
antisense molecule is 5'- ATCTCTCCGCTTCCTTTC -3' ( < 400 > 10).
Other particularly useful antisense molecules are:
#27 UCCGGAGCCAGACUU
#64 CACAGUUGCUGCAAG
#78 UCUCCGCUUCCUUUC
#28 AGCCCCCACAGCGAG
#32 GCCUUGGAGAUGAGC
#40 UAACAGAGGUCAGCA
#42 GGAUCAGGGACCAGU
#46 CGGCAAGCUACACAG
#50 GGCAGGCAGGCACAC
Particularly useful molecules are selected from #27, #64 and #78. In a
preferred embodiment
these molecules comprise a C-5 propynyl dU, dC phosphorothioate modification.
A particularly preferred embodiment of the present invention contemplates a
method of
ameliorating the effects of psoriasis or other medical disorder, said method
comprising
contacting proliferating skin or skin capable of proliferation or cells
associated with said
medical disorder with an effective amount of one or more nucleic acid
molecules or chemical
analogues thereof capable of inhibiting or otherwise reducing IGF-I mediated
cell proliferation
or ameliorating the medical disorder wherein said one or more molecules
comprises a
polynucleotide capable of interacting with mRNA directed from an IGF-I gene,
an IGF-I
receptor gene or a gene encoding an IGFBP such as IGFBP-2 and/or IGFBP-3.
Preferably, the nucleic acid molecule are antisense molecules. Particularly
useful antisense
molecules are:
#27 UCCGGAGCCAGACUU
#64 CACAGUUGCUGCAAG


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#78 UCUCCGCUUCCUUUC
#28 AGCCCCCACAGCGAG
#32 GCCUUGGAGAUGAGC
#40 UAACAGAGGUCAGCA
#42 GGAUCAGGGACCAGU
#46 CGGCAAGCUACACAG
#50 GGCAGGCAGGCACAC
Even more particularly useful molecules are selected from #27, #64 and #78.
In accordance with one aspect of the present invention the nucleic acid
molecule is topically
applied in aqueous solution or in conjunction with a cream, ointment, oil or
other suitable carrier
and/or diluent. A single application may be sufficient depending on the
severity or exigencies
of the condition although more commonly, multiple applications are required
ranging from
hourly, multi-hourly, daily, multi-daily, weekly or monthly, or in some other
suitable time
interval. The treatment might comprise solely the application of the nucleic
acid molecule or
this may be applied in conjunction with other treatments for the skin
proliferation and/or
inflammatory disorder being treated or for other associated conditions
including microbial
infection, bleeding and the formation of a variety of rashes.
As an alternative to or in conjunction with antisense therapy, the subject
invention extends to
the nucleic acid molecule as, or incorporating, a ribozyme including a
minizyme to, for
example, IGF-I, its receptor or to molecules such as IGFBPs and in particular
IGFBP-2 and -3.
Ribozymes are synthetic nucleic acid molecules which possess highly specific
endoribonuclease
activity. In particular, they comprise a hybridising region which is
complementary in nucleotide
sequence to at least part of a target RNA. Ribozymes are well described by
Haseloff and
Gerlach [8] and in International Patent Application No. WO 89/05852. The
present invention
extends to ribozymes which target mRNA specified by genes encoding IGF-I, its
receptor or one
or more IGFBPs such as IGFBP-2 and/or IGFBP-3.


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According to this embodiment, there is provided in a particularly preferred
aspect a ribozyme
comprising a hybridising region and a catalytic region wherein the hybridising
region is capable
of hybridising to at least part of a target mRNA sequence transcribed from a
genomic gene
corresponding to (<400>1) or (<400>2) wherein said catalytic domain is capable
of cleaving
S said target mRNA sequence to reduce or inhibit IGF-I mediated cell
proliferation and/or
inflammation and/or other medical disorders.
Yet another aspect of the present invention contemplates co-suppression to
reduce expression
or to inhibit translation of an endogenous gene encoding, for example, IGF-I,
its receptor, or
IGFBPs such as IGFBP-2 and/or -3. In co-suppression, a second copy of an
endogenous gene
or a substantially similar copy or analogue of an endogenous gene is
introduced into a cell
following topical administration. As with antisense molecules, nucleic acid
molecules defining
a ribozyme or nucleic acid molecules useful in co-suppression may first be
protected such as by
using a nonionic backbone.
The efficacy of the nucleic acid molecules of the present invention can be
conveniently tested
and screened using an in vitro system comprising a basal keratinocyte cell
line. A particularly
useful system comprises the HaCaT cell line described by Boukamp et al [9]. In
one assay,
IGF-I is added to an oligonucleotide treated HaCaT cell line. Alternatively,
growth of
oligonucleotide treated HaCaT cells is observed on a feeder layer of
irradiated 3T3 fibroblasts.
Using such in vitro assays, it is observed that antisense oligonucleotides to
IGFBP-3, for
example, inhibit production of IGFBP-3 by HaCaT cells. Other suitable animal
models
include the nude mouse/human skin graft model (15; 16) and the "flaky skin"
mouse model (17;
18). In the nude mouse model, microdermatome biopsies of psoriasis lesions are
taken under
local anaesthetic from volunteers then transplanted to congenital athymic
(nude) mice. These
transplanted human skin grafts maintain the characteristic hyperproliferating
epidermis for 6-8
weeks. They are an established model for testing the efficacy of topically
applied therapies for
psoriasis. In the "flaky skin" mouse model, the fsn/fsn mutation produces mice
with skin
resembling human psoriasis. This mouse, or another mutant mouse with a similar
phenotype
is a further in vivo model to test the efficacy of topically applied therapies
for psoriasis.


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Another aspect of the present invention contemplates a pharmaceutical
composition for topical
administration which comprises a nucleic acid molecule capable of inhibiting
or otherwise
reducing IGF-I mediated cell proliferation such as psoriasis and one or more
pharmaceutically
acceptable carriers and/or diluents. Preferably, the nucleic acid molecule is
an antisense
S molecule to IGF-I, the IGF-I receptor or an IGFBP such as IGFBP-2 and/or
IGFBP-3 or
comprises a ribozyme to one or more of these targets or is a molecule suitable
for co
suppression of one or more of these targets. The composition may comprise a
single species of
a nucleic acid molecule capable of targeting one of IGF-I, its receptor or an
IGFBP, such as
IGFBP-2 or IGFBP-3 or may be a multi-valent molecule capable of targeting two
or more of
IGF-I, its receptor or an IGFBP, such as IGFBP-2 and/or IGFBP-3.
The nucleic acid molecules may be administered in dispersions prepared in
creams, ointments,
oil or other suitable carrier and/or diluent such as glycerol, liquid
polyethylene glycols and/or
mixtures thereof. Under ordinary conditions of storage and use, these
preparations may contain
1 S a preservative to prevent the growth of microorganisms.
The pharmaceutical forms suitable for topical use include sterile aqueous
solutions (where water
soluble) or dispersions and powders for the extemporaneous preparation of
topical solutions or
dispersions. In all cases, the form is preferably sterile although this is not
an absolute
requirement and is stable under the conditions of manufacture and storage. The
carrier can be
a solvent or dispersion medium containing, for example, water, ethanol, polyol
(for example,
glycerol, propylene glycol, and liquid polyethylene glycol, and the like),
suitable mixtures
thereof and vegetable oils. The proper fluidity can be maintained, for
example, by the use of
a coating such as lecithin, by the maintenance of the required particle size
in the case of
dispersion and by the use of superfactants. The prevention of the action of
microorganism can
be brought about by various antibacterial and antifungal agents, for example,
parabens,
chlorobutanol, phenol, sorbic acid, thimerosal and the like. In many cases, it
will be preferable
to include isotonic agents, for example, sugars or sodium chloride.


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Topical solutions are prepared by incorporating the nucleic acid molecule
compound in the
required amount in the appropriate solvent with various of the other
ingredients enumerated
above, as required, followed by where necessary filter sterilization.
The active agent may alternatively be administered by intravenous,
subcutaneous, nasal drip,
suppository, implant means amongst other suitable routes of administration
including
intraperitoneal, intramuscular, absorption through epithelial or mucocutaneous
linings for
example via nasal, oral, vaginal, rectal or gastrointestinal administration.
Reference may
conveniently be made to reference 24.
As used herein "pharmaceutically acceptable carriers and/or diluents" include
any and all
solvents, dispersion media, aqueous solutions, coatings, antibacterial and
antifungal agents,
isotonic and absorption delaying agents, and the like. The use of such media
and agents for
pharmaceutical active substances is well known in the art. Except insofar as
any conventional
media or agent is incompatible with the active ingredient, use thereof in the
pharmaceutical
compositions is contemplated. Supplementary active ingredients can also be
incorporated into
the compositions. Conveniently, the nucleic acid molecules of the present
invention are stored
in freeze-dried form and are reconstituted prior to use.
Yet another aspect of the present invention contemplates the use of a nucleic
acid molecule in
the manufacture of a medicament for the treatment of proliferative and/or
inflammatory skin
disorders or other medical disorders mediated by a growth factor. The
proliferative and/or
inflammatory skin disorder is generally psoriasis or other medical disorders
as described above
and the nucleic acid molecule targets IGF-I, the IGF-I receptor and/or an
IGFBP such as IGFBP-
2 and/or IGFBP-3.
Still a further aspect of the present invention contemplates an agent
comprising a nucleic acid
molecule as hereinbefore defined useful in the treatment of proliferative
and/or inflammatory
skin disorders, such as psoriasis or other medical disorder..


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The present invention further contemplates the use of the genetic molecules
and in particular
the antisense molecules to inhibit the anti-apoptotic activity of IGF-I
receptor. Such a use is
appropriate for the treatment of certain cancers and as adjunct therapy for
epidermal hyperplasia
such as in combination with UV treatment.
The present invention is further described by the following non-limiting
Examples.


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EXAMPLE 1
The differentiated human keratinocyte cell line, HaCaT [9] was used in the in
vitro assay. Cells
at passage numbers 33 to 36 were maintained as monolayer cultures in 5% v/v
COZ at 37°C in
Keratinocyte-SFM (Gibco) containing EGF and bovine pituitary extract as
supplied. Media
containing foetal calf serum were avoided because of the high content of IGF-I
binding proteins
in serum.
Feeder layer plates of lethally irradiated 3T3 fibroblasts were prepared
exactly as described by
Rheinwald and Green [10].
EXAMPLE 2
Cells were grown to 4 days post confluence in 2cm2 wells with daily medium
changes of
Keratinocyte-SFM, then the medium was changed to DMEM (Cytosystems,
Australia), with the
following additions: 25mM Hepes, 0.19% w/v, sodium bicarbonate, 0.03% w/v
glutamine
(Sigma Chemical Co, USA), SOILT/ml penicillin and SO~g/ml streptomycin (Flow
Laboratories).
After 24 hours, IGF-I or tIGF-I was added to triplicate wells, at the
concentrations indicated, in
O.SmI fresh DMEM containing 0.02% v/v bovine serum albumin (Sigma molecular
biology
grade) and incubated for a further 21 hours. [3H]-Thymidine (0.1 ~.Ci/well)
was then added and
the cells incubated for a further 3 hours. The medium was then aspirated and
the cells washed
once with ice-cold PBS and twice with ice-cold 10% v/v TCA. The TCA-
precipitated
monolayers were then solubilized with 0.25M NaOH (200~1/well), transferred to
scintillation
vials and radioactivity determined by liquid scintillation counting (Pharmacia
Wallac 1410
liquid scintillation counter).
EXAMPLE 3
HaCaT conditioned medium (2501) was concentrated by adding 7501 cold ethanol,
incubating
at -20°C for 2 hours and centrifuging at 16,OOOg for 20 min at
4°C. The resulting pellet was
air dried, resuspended thoroughly in non-reducing Laemmli sample buffer,
heated to 90°C for
5 minutes and separated on 12% w/v SDS-PAGE according to the method of Laemmli
(1970).
Separated proteins were electrophoretically transferred to nitrocellulose
membrane (0.45mm,
Schleicher and Schuell, Dassel, Germany) in a buffer containing 25mM Tris,
192mM glycine


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and 20% v/v methanol. IGFBPs were then visualised by the procedure of
Hossenlopp et al [ 11 ],
using ['ZSI]-IGF-I, followed by autoradiography. Autoradiographs were scanned
in a BioRad
Model GS-670 Imaging Densitometer and band densities were determined using the
Molecular
Analyst program.
EXAMPLE 4
Phosphorothioate oligodeoxynucleotides were synthesised by Bresatec, Adelaide,
South
Australia, Australia. The following antisense sequences were used: BP3AS2, 5'-
GCG CCC
GCT GCA TGA CGC CTG CAA C -3' (<400>4), a 25mer complementary to the start
codon
region of the human IGFBP-3 mRNA; BP3AS3, 5'- CGG GCG GCT CAC CTG GAG CTG
GCG -3' (<400>5), a 24mer complementary to the exon 1/intron 1 splice site;
BP3AS4, 5'-
AGG CGG CTG ACG GCA CTA -3'(<400>6), an l8mer complementary to a region of the
coding sequence lacking RNA secondary structure and oligonucleotide-dimer
formation (using
the computer software "OLIGO for PC"). Since BP3AS4 was found to be
ineffective at
inhibiting IGFBP-3 synthesis, it was used as a control. The following
additional control
oligonucleotide sequences were used: BP3S, 5'- CAG GCG TCA TGC AGC GGG C -3'
(<400>7), an l8mer sense control sequence equivalent to the start codon
region; BP3AS2NS,
S'- CGG AGA TGC CGC ATG CCA GCG CAG G -3' (<400>8), a 25mer randomised
sequence with the same GC content as BP3AS2; BP3AS4NS, 5'- GAC AGC GTC GGA GCG
ATC -3' (<400>9), an l8mer randomised sequence with the same GC content as
BP3AS4NS.
Design of the oligonucleotides was based on the human IGFBP-3 cDNA sequence of
Spratt et
al [12].
Cells were grown to one day post confluence in 2cm2 wells with daily medium
changes of O.SmI
Keratinocyte-SFM, then subjected to daily medium changes of Keratinocyte-SFM
for a further
4 days. Daily additions of O.SmI fresh Keratinocyte-SFM were then continued
for a further 7
days, except that at the time of medium addition, 5~1 oligonucleotide in PBS
was added to give
the final concentrations indicated, then the wells were shaken to mix the
oligonucleotide. After
the final addition, cells were incubated for 24 hours and the medium collected
for assay of
IGFBPs. Cells were then counted after trypsinisation in a Coulter Industrial D
Counter, Coulter
Bedfordshire, UK. Cell numbers after oligonucleotide treatment differed by
less than 10%.


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EXAMPLE 5
HaCaT cells secrete mainly IGFBP-3 (>95%), with the only other IGFBP
detectable in HaCaT
conditioned medium being IGFBP-4 (<5%). The effect on IGFBP-3 and IGFBP-4
synthesis of
antisense oligonucleotides at two concentrations, SpM and O.SpM, was tested.
Two
S oligonucleotides were used, BP3AS2 and BP3AS3, directed against the start
site and the intron
1/exon 1 splice site, respectively of the IGFBP-3 mRNA. As a control, a sense
oligonucleotide
corresponding to the start site was used. As shown in Figures 4A and 4B, all
oligonucleotides
at SpM caused a significant reduction of IGFBP-3 synthesis compared with
untreated cells,
however, the two antisense oligonucleotides inhibited IGFBP-3 synthesis of
approximately SO%
compared to the sense control (Figure 4B). The antisense oligonucleotide
directed to the start
codon appeared to be more effective of the two, the difference being more
apparent at the lower
concentration of O.SpM. The cells of IGFBP-4 secreted by the HaCaT cells make
photographic
reproduction of the bands on Western ligand blots difficult, however
densitometry
measurements provide adequate relative quantitation. This resulted in the
significant
observation that IGFBP-4 levels were unaffected by oligonucleotide addition to
the cells,
suggesting that the observed inhibitory effects on IGFBP-3 are specific.
To further investigate the inhibitory effects of the more effective of the two
antisense
oligonucleotides, BP3AS2, inhibition by this oligonucleotide at O.SpM was
compared with a
number of control oligonucleotides, including one antisense oligonucleotide to
IGFBP-3 that
had proved to be ineffective at O.SpM. As shown in Figures SA and SB, BP3AS2
was again
inhibitory, resulting in levels of IGFBP-3 of approximately SO% of the most
non-specifically
inhibitory control oligonucleotide, the randomised equivalent of BP3AS2. The
other control
oligonucleotides caused no reduction in IGFBP-3 levels at O.SpM, compared to
untreated cells.
Of possible significance is the fact that this control oligonucleotide,
BP3AS2NS, like BP3AS2
itself, has the highest potential Tm of the three control oligonucleotides
used in this experiment,
enhancing the probability of non-specific base pairing with non-target mRNAs.
However, the
lack of inhibition of IGFBP-4 secretion by BP3AS2 suggests that this
oligonucleotide is
selective even compared with the most closely related protein likely to be
present in this cell
line.


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EXAMPLE 6
Antisense oligonucleotides to IGFBP2 may be selected from molecules capable of
interacting
with one or more of the following sense oligonucleotides:
ATTCGGGGCGAGGGA CGCAGGGCCGTGCAC CCGCGCCGCGCTGCC


TTCGGGGCGAGGGAG GCAGGGCCGTGCACC CGCGCCGCGCTGCCG


TCGGGGCGAGGGAGG CAGGGCCGTGCACCT GCGCCGCGCTGCCGA


CGGGGCGAGGGAGGA AGGGCCGTGCACCTG CGCCGCGCTGCCGAC


GGGGCGAGGGAGGAG GGGCCGTGCACCTGC GCCGCGCTGCCGACC


GGGCGAGGGAGGAGG GGCCGTGCACCTGCC CCGCGCTGCCGACCG


10GGCGAGGGAGGAGGA GCCGTGCACCTGCCC CGCGCTGCCGACCGC


GCGAGGGAGGAGGAA CCGTGCACCTGCCCG GCGCTGCCGACCGCC


CGAGGGAGGAGGAAG CGTGCACCTGCCCGC CGCTGCCGACCGCCA


GAGGGAGGAGGAAGA GTGCACCTGCCCGCC GCTGCCGACCGCCAG


AGGGAGGAGGAAGAA TGCACCTGCCCGCCC CTGCCGACCGCCAGC


15GGGAGGAGGAAGAAG GCACCTGCCCGCCCG TGCCGACCGCCAGCA


GGAGGAGGAAGAAGC CACCTGCCCGCCCGC GCCGACCGCCAGCAT


GAGGAGGAAGAAGCG ACCTGCCCGCCCGCC CCGACCGCCAGCATG


AGGAGGAAGAAGCGG CCTGCCCGCCCGCCC CGACCGCCAGCATGC


GGAGGAAGAAGCGGA CTGCCCGCCCGCCCG GACCGCCAGCATGCT


20GAGGAAGAAGCGGAG TGCCCGCCCGCCCGC ACCGCCAGCATGCTG


AGGAAGAAGCGGAGG GCCCGCCCGCCCGCT CCGCCAGCATGCTGC


GGAAGAAGCGGAGGA CCCGCCCGCCCGCTC CGCCAGCATGCTGCC


GAAGAAGCGGAGGAG CCGCCCGCCCGCTCG GCCAGCATGCTGCCG


AAGAAGCGGAGGAGG CGCCCGCCCGCTCGC CCAGCATGCTGCCGA


25AGAAGCGGAGGAGGC GCCCGCCCGCTCGCT CAGCATGCTGCCGAG


GAAGCGGAGGAGGCG CCCGCCCGCTCGCTC AGCATGCTGCCGAGA


AAGCGGAGGAGGCGG CCGCCCGCTCGCTCG GCATGCTGCCGAGAG


AGCGGAGGAGGCGGC CGCCCGCTCGCTCGC CATGCTGCCGAGAGT


GCGGAGGAGGCGGCT GCCCGCTCGCTCGCT ATGCTGCCGAGAGTG


30CGGAGGAGGCGGCTC CCCGCTCGCTCGCTC TGCTGCCGAGAGTGG


GGAGGAGGCGGCTCC CCGCTCGCTCGCTCG GCTGCCGAGAGTGGG


GAGGAGGCGGCTCCC CGCTCGCTCGCTCGC CTGCCGAGAGTGGGC


AGGAGGCGGCTCCCG GCTCGCTCGCTCGCC TGCCGAGAGTGGGCT


GGAGGCGGCTCCCGC CTCGCTCGCTCGCCC GCCGAGAGTGGGCTG


35GAGGCGGCTCCCGCT TCGCTCGCTCGCCCG CCGAGAGTGGGCTGC


AGGCGGCTCCCGCTC CGCTCGCTCGCCCGC CGAGAGTGGGCTGCC


GGCGGCTCCCGCTCG GCTCGCTCGCCCGCC GAGAGTGGGCTGCCC


GCGGCTCCCGCTCGC CTCGCTCGCCCGCCG AGAGTGGGCTGCCCC


CGGCTCCCGCTCGCA TCGCTCGCCCGCCGC GAGTGGGCTGCCCCG


40GGCTCCCGCTCGCAG CGCTCGCCCGCCGCG AGTGGGCTGCCCCGC


GCTCCCGCTCGCAGG GCTCGCCCGCCGCGC GTGGGCTGCCCCGCG


CTCCCGCTCGCAGGG CTCGCCCGCCGCGCC TGGGCTGCCCCGCGC


TCCCGCTCGCAGGGC TCGCCCGCCGCGCCG GGGCTGCCCCGCGCT


CCCGCTCGCAGGGCC CGCCCGCCGCGCCGC GGCTGCCCCGCGCTG


45CCGCTCGCAGGGCCG GCCCGCCGCGCCGCG GCTGCCCCGCGCTGC


CGCTCGCAGGGCCGT CCCGCCGCGCCGCGC CTGCCCCGCGCTGCC


GCTCGCAGGGCCGTG CCGCCGCGCCGCGCT TGCCCCGCGCTGCCG


CTCGCAGGGCCGTGC CGCCGCGCCGCGCTG GCCCCGCGCTGCCGC


TCGCAGGGCCGTGCA GCCGCGCCGCGCTGC CCCCGCGCTGCCGCT




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CCCGCGCTGCCGCTG CTGCTGCTACTGGGC CTGTTCCGCTGCCCG


CCGCGCTGCCGCTGC TGCTGCTACTGGGCG TGTTCCGCTGCCCGC


CGCGCTGCCGCTGCC GCTGCTACTGGGCGC GTTCCGCTGCCCGCC


GCGCTGCCGCTGCCG CTGCTACTGGGCGCG TTCCGCTGCCCGCCC


CGCTGCCGCTGCCGC TGCTACTGGGCGCGA TCCGCTGCCCGCCCT


GCTGCCGCTGCCGCC GCTACTGGGCGCGAG CCGCTGCCCGCCCTG


CTGCCGCTGCCGCCG CTACTGGGCGCGAGT CGCTGCCCGCCCTGC


TGCCGCTGCCGCCGC TACTGGGCGCGAGTG GCTGCCCGCCCTGCA


GCCGCTGCCGCCGCC ACTGGGCGCGAGTGG CTGCCCGCCCTGCAC


10CCGCTGCCGCCGCCG CTGGGCGCGAGTGGC TGCCCGCCCTGCACA


CGCTGCCGCCGCCGC TGGGCGCGAGTGGCG GCCCGCCCTGCACAC


GCTGCCGCCGCCGCC GGGCGCGAGTGGCGG CCCGCCCTGCACACC


CTGCCGCCGCCGCCG GGCGCGAGTGGCGGC CCGCCCTGCACACCC


TGCCGCCGCCGCCGC GCGCGAGTGGCGGCG CGCCCTGCACACCCG


15GCCGCCGCCGCCGCT CGCGAGTGGCGGCGG GCCCTGCACACCCGA


CCGCCGCCGCCGCTG GCGAGTGGCGGCGGC CCCTGCACACCCGAG


CGCCGCCGCCGCTGC CGAGTGGCGGCGGCG CCTGCACACCCGAGC


GCCGCCGCCGCTGCT GAGTGGCGGCGGCGG CTGCACACCCGAGCG


CCGCCGCCGCTGCTG AGTGGCGGCGGCGGC TGCACACCCGAGCGC


20CGCCGCCGCTGCTGC GTGGCGGCGGCGGCG GCACACCCGAGCGCC


GCCGCCGCTGCTGCC TGGCGGCGGCGGCGG CACACCCGAGCGCCT


CCGCCGCTGCTGCCG GGCGGCGGCGGCGGG ACACCCGAGCGCCTG


CGCCGCTGCTGCCGC GCGGCGGCGGCGGGG CACCCGAGCGCCTGG


GCCGCTGCTGCCGCT CGGCGGCGGCGGGGC ACCCGAGCGCCTGGC


25CCGCTGCTGCCGCTG GGCGGCGGCGGGGCG CCCGAGCGCCTGGCC


CGCTGCTGCCGCTGC GCGGCGGCGGGGCGC CCGAGCGCCTGGCCG


GCTGCTGCCGCTGCT CGGCGGCGGGGCGCG CGAGCGCCTGGCCGC


CTGCTGCCGCTGCTG GGCGGCGGGGCGCGC GAGCGCCTGGCCGCC


TGCTGCCGCTGCTGC GCGGCGGGGCGCGCG AGCGCCTGGCCGCCT


30GCTGCCGCTGCTGCC CGGCGGGGCGCGCGC GCGCCTGGCCGCCTG


CTGCCGCTGCTGCCG ' GGCGGGGCGCGCGCG CGCCTGGCCGCCTGC


TGCCGCTGCTGCCGC GCGGGGCGCGCGCGG GCCTGGCCGCCTGCG


GCCGCTGCTGCCGCT CGGGGCGCGCGCGGA CCTGGCCGCCTGCGG


CCGCTGCTGCCGCTG GGGGCGCGCGCGGAG CTGGCCGCCTGCGGG


35CGCTGCTGCCGCTGC GGGCGCGCGCGGAGG TGGCCGCCTGCGGGC


GCTGCTGCCGCTGCT GGCGCGCGCGGAGGT GGCCGCCTGCGGGCC


CTGCTGCCGCTGCTG GCGCGCGCGGAGGTG GCCGCCTGCGGGCCC


TGCTGCCGCTGCTGC CGCGCGCGGAGGTGC CCGCCTGCGGGCCCC


GCTGCCGCTGCTGCT GCGCGCGGAGGTGCT CGCCTGCGGGCCCCC


40CTGCCGCTGCTGCTG CGCGCGGAGGTGCTG GCCTGCGGGCCCCCG


TGCCGCTGCTGCTGC GCGCGGAGGTGCTGT CCTGCGGGCCCCCGC


GCCGCTGCTGCTGCT CGCGGAGGTGCTGTT CTGCGGGCCCCCGCC


CCGCTGCTGCTGCTG GCGGAGGTGCTGTTC TGCGGGCCCCCGCCG


CGCTGCTGCTGCTGC CGGAGGTGCTGTTCC GCGGGCCCCCGCCGG


45GCTGCTGCTGCTGCT GGAGGTGCTGTTCCG CGGGCCCCCGCCGGT


CTGCTGCTGCTGCTA GAGGTGCTGTTCCGC GGGCCCCCGCCGGTT


TGCTGCTGCTGCTAC AGGTGCTGTTCCGCT GGCCCCCGCCGGTTG


GCTGCTGCTGCTACT GGTGCTGTTCCGCTG GCCCCCGCCGGTTGC


CTGCTGCTGCTACTG GTGCTGTTCCGCTGC CCCCCGCCGGTTGCG


50TGCTGCTGCTACTGG TGCTGTTCCGCTGCC CCCCGCCGGTTGCGC


GCTGCTGCTACTGGG GCTGTTCCGCTGCCC CCCGCCGGTTGCGCC




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CCGCCGGTTGCGCCG ATGCCATGCGCGGAG TGCGCCCGGCTGGAG


CGCCGGTTGCGCCGC TGCCATGCGCGGAGC GCGCCCGGCTGGAGG


GCCGGTTGCGCCGCC GCCATGCGCGGAGCT CGCCCGGCTGGAGGG


CCGGTTGCGCCGCCC CCATGCGCGGAGCTC GCCCGGCTGGAGGGC


CGGTTGCGCCGCCCG CATGCGCGGAGCTCG CCCGGCTGGAGGGCG


GGTTGCGCCGCCCGC ATGCGCGGAGCTCGT CCGGCTGGAGGGCGA


GTTGCGCCGCCCGCC TGCGCGGAGCTCGTC CGGCTGGAGGGCGAG


TTGCGCCGCCCGCCG GCGCGGAGCTCGTCC GGCTGGAGGGCGAGG


TGCGCCGCCCGCCGC CGCGGAGCTCGTCCG GCTGGAGGGCGAGGC


10GCGCCGCCCGCCGCG GCGGAGCTCGTCCGG CTGGAGGGCGAGGCG


CGCCGCCCGCCGCGG CGGAGCTCGTCCGGG TGGAGGGCGAGGCGT


GCCGCCCGCCGCGGT GGAGCTCGTCCGGGA GGAGGGCGAGGCGTG


CCGCCCGCCGCGGTG GAGCTCGTCCGGGAG GAGGGCGAGGCGTGC


CGCCCGCCGCGGTGG AGCTCGTCCGGGAGC AGGGCGAGGCGTGCG


15GCCCGCCGCGGTGGC GCTCGTCCGGGAGCC GGGCGAGGCGTGCGG


CCCGCCGCGGTGGCC CTCGTCCGGGAGCCG GGCGAGGCGTGCGGC


CCGCCGCGGTGGCCG TCGTCCGGGAGCCGG GCGAGGCGTGCGGCG


CGCCGCGGTGGCCGC CGTCCGGGAGCCGGG CGAGGCGTGCGGCGT


GCCGCGGTGGCCGCA GTCCGGGAGCCGGGC GAGGCGTGCGGCGTC


20CCGCGGTGGCCGCAG TCCGGGAGCCGGGCT AGGCGTGCGGCGTCT


CGCGGTGGCCGCAGT CCGGGAGCCGGGCTG GGCGTGCGGCGTCTA


GCGGTGGCCGCAGTG CGGGAGCCGGGCTGC GCGTGCGGCGTCTAC


CGGTGGCCGCAGTGG GGGAGCCGGGCTGCG CGTGCGGCGTCTACA


GGTGGCCGCAGTGGC GGAGCCGGGCTGCGG GTGCGGCGTCTACAC


25GTGGCCGCAGTGGCC GAGCCGGGCTGCGGC TGCGGCGTCTACACC


TGGCCGCAGTGGCCG AGCCGGGCTGCGGCT GCGGCGTCTACACCC


GGCCGCAGTGGCCGG GCCGGGCTGCGGCTG CGGCGTCTACACCCC


GCCGCAGTGGCCGGA CCGGGCTGCGGCTGC GGCGTCTACACCCCG


CCGCAGTGGCCGGAG CGGGCTGCGGCTGCT GCGTCTACACCCCGC


30CGCAGTGGCCGGAGG GGGCTGCGGCTGCTG CGTCTACACCCCGCG


GCAGTGGCCGGAGGC GGCTGCGGCTGCTGC GTCTACACCCCGCGC


CAGTGGCCGGAGGCG GCTGCGGCTGCTGCT TCTACACCCCGCGCT


AGTGGCCGGAGGCGC CTGCGGCTGCTGCTC CTACACCCCGCGCTG


GTGGCCGGAGGCGCC TGCGGCTGCTGCTCG TACACCCCGCGCTGC


35TGGCCGGAGGCGCCC GCGGCTGCTGCTCGG ACACCCCGCGCTGCG


GGCCGGAGGCGCCCG CGGCTGCTGCTCGGT CACCCCGCGCTGCGG


GCCGGAGGCGCCCGC GGCTGCTGCTCGGTG ACCCCGCGCTGCGGC


CCGGAGGCGCCCGCA GCTGCTGCTCGGTGT CCCCGCGCTGCGGCC


CGGAGGCGCCCGCAT CTGCTGCTCGGTGTG CCCGCGCTGCGGCCA


40GGAGGCGCCCGCATG TGCTGCTCGGTGTGC CCGCGCTGCGGCCAG


GAGGCGCCCGCATGC GCTGCTCGGTGTGCG CGCGCTGCGGCCAGG


AGGCGCCCGCATGCC CTGCTCGGTGTGCGC GCGCTGCGGCCAGGG


GGCGCCCGCATGCCA TGCTCGGTGTGCGCC CGCTGCGGCCAGGGG


GCGCCCGCATGCCAT GCTCGGTGTGCGCCC GCTGCGGCCAGGGGC


45CGCCCGCATGCCATG CTCGGTGTGCGCCCG CTGCGGCCAGGGGCT


GCCCGCATGCCATGC TCGGTGTGCGCCCGG TGCGGCCAGGGGCTG


CCCGCATGCCATGCG CGGTGTGCGCCCGGC GCGGCCAGGGGCTGC


CCGCATGCCATGCGC GGTGTGCGCCCGGCT CGGCCAGGGGCTGCG


CGCATGCCATGCGCG GTGTGCGCCCGGCTG GGCCAGGGGCTGCGC


50GCATGCCATGCGCGG TGTGCGCCCGGCTGG GCCAGGGGCTGCGCT


CATGCCATGCGCGGA GTGCGCCCGGCTGGA CCAGGGGCTGCGCTG




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CAGGGGCTGCGCTGC CTGGTCATGGGCGAG GCCAGCCCGGAGCAG


AGGGGCTGCGCTGCT TGGTCATGGGCGAGG CCAGCCCGGAGCAGG


GGGGCTGCGCTGCTA GGTCATGGGCGAGGG CAGCCCGGAGCAGGT


GGGCTGCGCTGCTAT GTCATGGGCGAGGGC AGCCCGGAGCAGGTT


GGCTGCGCTGCTATC TCATGGGCGAGGGCA GCCCGGAGCAGGTTG


GCTGCGCTGCTATCC CATGGGCGAGGGCAC CCCGGAGCAGGTTGC


CTGCGCTGCTATCCC ATGGGCGAGGGCACT CCGGAGCAGGTTGCA


TGCGCTGCTATCCCC TGGGCGAGGGCACTT CGGAGCAGGTTGCAG


GCGCTGCTATCCCCA GGGCGAGGGCACTTG GGAGCAGGTTGCAGA


10CGCTGCTATCCCCAC GGCGAGGGCACTTGT GAGCAGGTTGCAGAC


GCTGCTATCCCCACC GCGAGGGCACTTGTG AGCAGGTTGCAGACA


CTGCTATCCCCACCC CGAGGGCACTTGTGA GCAGGTTGCAGACAA


TGCTATCCCCACCCG GAGGGCACTTGTGAG CAGGTTGCAGACAAT


GCTATCCCCACCCGG AGGGCACTTGTGAGA AGGTTGCAGACAATG


15CTATCCCCACCCGGG GGGCACTTGTGAGAA GGTTGCAGACAATGG


TATCCCCACCCGGGC GGCACTTGTGAGAAG GTTGCAGACAATGGC
.


ATCCCCACCCGGGCT GCACTTGTGAGAAGC TTGCAGACAATGGCG


TCCCCACCCGGGCTC CACTTGTGAGAAGCG TGCAGACAATGGCGA


CCCCACCCGGGCTCC ACTTGTGAGAAGCGC GCAGACAATGGCGAT


20CCCACCCGGGCTCCG CTTGTGAGAAGCGCC CAGACAATGGCGATG


CCACCCGGGCTCCGA TTGTGAGAAGCGCCG AGACAATGGCGATGA


CACCCGGGCTCCGAG TGTGAGAAGCGCCGG GACAATGGCGATGAC


ACCCGGGCTCCGAGC GTGAGAAGCGCCGGG ACAATGGCGATGACC


CCCGGGCTCCGAGCT TGAGAAGCGCCGGGA CAATGGCGATGACCA


25CCGGGCTCCGAGCTG GAGAAGCGCCGGGAC AATGGCGATGACCAC


CGGGCTCCGAGCTGC AGAAGCGCCGGGACG ATGGCGATGACCACT


GGGCTCCGAGCTGCC GAAGCGCCGGGACGC TGGCGATGACCACTC


GGCTCCGAGCTGCCC AAGCGCCGGGACGCC GGCGATGACCACTCA


GCTCCGAGCTGCCCC AGCGCCGGGACGCCG GCGATGACCACTCAG


30CTCCGAGCTGCCCCT GCGCCGGGACGCCGA CGATGACCACTCAGA


TCCGAGCTGCCCCTG CGCCGGGACGCCGAG GATGACCACTCAGAA


CCGAGCTGCCCCTGC GCCGGGACGCCGAGT ATGACCACTCAGAAG


CGAGCTGCCCCTGCA CCGGGACGCCGAGTA TGACCACTCAGAAGG


GAGCTGCCCCTGCAG CGGGACGCCGAGTAT GACCACTCAGAAGGA


35AGCTGCCCCTGCAGG GGGACGCCGAGTATG ACCACTCAGAAGGAG


GCTGCCCCTGCAGGC GGACGCCGAGTATGG CCACTCAGAAGGAGG


CTGCCCCTGCAGGCG GACGCCGAGTATGGC CACTCAGAAGGAGGC


TGCCCCTGCAGGCGC ACGCCGAGTATGGCG ACTCAGAAGGAGGCC


GCCCCTGCAGGCGCT CGCCGAGTATGGCGC CTCAGAAGGAGGCCT


40CCCCTGCAGGCGCTG GCCGAGTATGGCGCC TCAGAAGGAGGCCTG


CCCTGCAGGCGCTGG CCGAGTATGGCGCCA CAGAAGGAGGCCTGG


CCTGCAGGCGCTGGT CGAGTATGGCGCCAG AGAAGGAGGCCTGGT


CTGCAGGCGCTGGTC GAGTATGGCGCCAGC GAAGGAGGCCTGGTG


TGCAGGCGCTGGTCA AGTATGGCGCCAGCC AAGGAGGCCTGGTGG


45GCAGGCGCTGGTCAT GTATGGCGCCAGCCC AGGAGGCCTGGTGGA


CAGGCGCTGGTCATG TATGGCGCCAGCCCG GGAGGCCTGGTGGAG


AGGCGCTGGTCATGG ATGGCGCCAGCCCGG GAGGCCTGGTGGAGA


GGCGCTGGTCATGGG TGGCGCCAGCCCGGA AGGCCTGGTGGAGAA


GCGCTGGTCATGGGC GGCGCCAGCCCGGAG GGCCTGGTGGAGAAC


50CGCTGGTCATGGGCG GCGCCAGCCCGGAGC GCCTGGTGGAGAACC


GCTGGTCATGGGCGA CGCCAGCCCGGAGCA CCTGGTGGAGAACCA




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CTGGTGGAGAACCAC AGTGCTGGCCGGAAG CGGGAGAAGGTCACT


TGGTGGAGAACCACG GTGCTGGCCGGAAGC GGGAGAAGGTCACTG


GGTGGAGAACCACGT TGCTGGCCGGAAGCC GGAGAAGGTCACTGA


GTGGAGAACCACGTG GCTGGCCGGAAGCCC GAGAAGGTCACTGAG


TGGAGAACCACGTGG CTGGCCGGAAGCCCC AGAAGGTCACTGAGC


GGAGAACCACGTGGA TGGCCGGAAGCCCCT GAAGGTCACTGAGCA


GAGAACCACGTGGAC GGCCGGAAGCCCCTC AAGGTCACTGAGCAG


AGAACCACGTGGACA GCCGGAAGCCCCTCA AGGTCACTGAGCAGC


GAACCACGTGGACAG CCGGAAGCCCCTCAA GGTCACTGAGCAGCA


10AACCACGTGGACAGC CGGAAGCCCCTCAAG GTCACTGAGCAGCAC


ACCACGTGGACAGCA GGAAGCCCCTCAAGT TCACTGAGCAGCACC


CCACGTGGACAGCAC GAAGCCCCTCAAGTC CACTGAGCAGCACCG


CACGTGGACAGCACC AAGCCCCTCAAGTCG ACTGAGCAGCACCGG


ACGTGGACAGCACCA AGCCCCTCAAGTCGG CTGAGCAGCACCGGC


15CGTGGACAGCACCAT GCCCCTCAAGTCGGG TGAGCAGCACCGGCA


GTGGACAGCACCATG CCCCTCAAGTCGGGT GAGCAGCACCGGCAG


TGGACAGCACCATGA CCCTCAAGTCGGGTA AGCAGCACCGGCAGA


GGACAGCACCATGAA CCTCAAGTCGGGTAT GCAGCACCGGCAGAT


GACAGCACCATGAAC CTCAAGTCGGGTATG CAGCACCGGCAGATG


20ACAGCACCATGAACA TCAAGTCGGGTATGA AGCACCGGCAGATGG


CAGCACCATGAACAT CAAGTCGGGTATGAA GCACCGGCAGATGGG


AGCACCATGAACATG AAGTCGGGTATGAAG CACCGGCAGATGGGC


GCACCATGAACATGT AGTCGGGTATGAAGG ACCGGCAGATGGGCA


CACCATGAACATGTT GTCGGGTATGAAGGA CCGGCAGATGGGCAA


25ACCATGAACATGTTG TCGGGTATGAAGGAG CGGCAGATGGGCAAG


CCATGAACATGTTGG CGGGTATGAAGGAGC GGCAGATGGGCAAGG


CATGAACATGTTGGG GGGTATGAAGGAGCT GCAGATGGGCAAGGG


ATGAACATGTTGGGC GGTATGAAGGAGCTG CAGATGGGCAAGGGT


TGAACATGTTGGGCG GTATGAAGGAGCTGG AGATGGGCAAGGGTG


30GAACATGTTGGGCGG TATGAAGGAGCTGGC GATGGGCAAGGGTGG


AACATGTTGGGCGGG ATGAAGGAGCTGGCC ATGGGCAAGGGTGGC


ACATGTTGGGCGGGG TGAAGGAGCTGGCCG TGGGCAAGGGTGGCA


CATGTTGGGCGGGGG GAAGGAGCTGGCCGT GGGCAAGGGTGGCAA


ATGTTGGGCGGGGGA AAGGAGCTGGCCGTG GGCAAGGGTGGCAAG


35TGTTGGGCGGGGGAG AGGAGCTGGCCGTGT GCAAGGGTGGCAAGC


GTTGGGCGGGGGAGG GGAGCTGGCCGTGTT CAAGGGTGGCAAGCA


TTGGGCGGGGGAGGC GAGCTGGCCGTGTTC AAGGGTGGCAAGCAT


TGGGCGGGGGAGGCA AGCTGGCCGTGTTCC AGGGTGGCAAGCATC


GGGCGGGGGAGGCAG GCTGGCCGTGTTCCG GGGTGGCAAGCATCA


40GGCGGGGGAGGCAGT CTGGCCGTGTTCCGG GGTGGCAAGCATCAC


GCGGGGGAGGCAGTG TGGCCGTGTTCCGGG GTGGCAAGCATCACC


CGGGGGAGGCAGTGC GGCCGTGTTCCGGGA TGGCAAGCATCACCT


GGGGGAGGCAGTGCT GCCGTGTTCCGGGAG GGCAAGCATCACCTT


GGGGAGGCAGTGCTG CCGTGTTCCGGGAGA GCAAGCATCACCTTG


45GGGAGGCAGTGCTGG CGTGTTCCGGGAGAA CAAGCATCACCTTGG


GGAGGCAGTGCTGGC GTGTTCCGGGAGAAG AAGCATCACCTTGGC


GAGGCAGTGCTGGCC TGTTCCGGGAGAAGG AGCATCACCTTGGCC


AGGCAGTGCTGGCCG GTTCCGGGAGAAGGT GCATCACCTTGGCCT


GGCAGTGCTGGCCGG TTCCGGGAGAAGGTC CATCACCTTGGCCTG


50GCAGTGCTGGCCGGA TCCGGGAGAAGGTCA ATCACCTTGGCCTGG


CAGTGCTGGCCGGAA CCGGGAGAAGGTCAC TCACCTTGGCCTGGA




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CACCTTGGCCTGGAG CCCTGCCAACAGGAA CTTCCGGATGAGCGG


ACCTTGGCCTGGAGG CCTGCCAACAGGAAC TTCCGGATGAGCGGG


CCTTGGCCTGGAGGA CTGCCAACAGGAACT TCCGGATGAGCGGGG


CTTGGCCTGGAGGAG TGCCAACAGGAACTG CCGGATGAGCGGGGC


TTGGCCTGGAGGAGC GCCAACAGGAACTGG CGGATGAGCGGGGCC


TGGCCTGGAGGAGCC CCAACAGGAACTGGA GGATGAGCGGGGCCC


GGCCTGGAGGAGCCC CAACAGGAACTGGAC GATGAGCGGGGCCCT


GCCTGGAGGAGCCCA AACAGGAACTGGACC ATGAGCGGGGCCCTC


CCTGGAGGAGCCCAA ACAGGAACTGGACCA TGAGCGGGGCCCTCT


10CTGGAGGAGCCCAAG CAGGAACTGGACCAG GAGCGGGGCCCTCTG


TGGAGGAGCCCAAGA AGGAACTGGACCAGG AGCGGGGCCCTCTGG


GGAGGAGCCCAAGAA GGAACTGGACCAGGT GCGGGGCCCTCTGGA


GAGGAGCCCAAGAAG GAACTGGACCAGGTC CGGGGCCCTCTGGAG


AGGAGCCCAAGAAGC AACTGGACCAGGTCC GGGGCCCTCTGGAGC


15GGAGCCCAAGAAGCT ACTGGACCAGGTCCT GGGCCCTCTGGAGCA


GAGCCCAAGAAGCTG CTGGACCAGGTCCTG GGCCCTCTGGAGCAC


AGCCCAAGAAGCTGC TGGACCAGGTCCTGG GCCCTCTGGAGCACC


GCCCAAGAAGCTGCG GGACCAGGTCCTGGA CCCTCTGGAGCACCT


CCCAAGAAGCTGCGA GACCAGGTCCTGGAG CCTCTGGAGCACCTC


20CCAAGAAGCTGCGAC ACCAGGTCCTGGAGC CTCTGGAGCACCTCT


CAAGAAGCTGCGACC CCAGGTCCTGGAGCG TCTGGAGCACCTCTA


AAGAAGCTGCGACCA CAGGTCCTGGAGCGG CTGGAGCACCTCTAC


AGAAGCTGCGACCAC AGGTCCTGGAGCGGA TGGAGCACCTCTACT


GAAGCTGCGACCACC GGTCCTGGAGCGGAT GGAGCACCTCTACTC


25AAGCTGCGACCACCC GTCCTGGAGCGGATC GAGCACCTCTACTCC


AGCTGCGACCACCCC TCCTGGAGCGGATCT AGCACCTCTACTCCC


GCTGCGACCACCCCC CCTGGAGCGGATCTC GCACCTCTACTCCCT


CTGCGACCACCCCCT CTGGAGCGGATCTCC CACCTCTACTCCCTG


TGCGACCACCCCCTG TGGAGCGGATCTCCA ACCTCTACTCCCTGC


30GCGACCACCCCCTGC GGAGCGGATCTCCAC CCTCTACTCCCTGCA


CGACCACCCCCTGCC GAGCGGATCTCCACC CTCTACTCCCTGCAC


GACCACCCCCTGCCA AGCGGATCTCCACCA TCTACTCCCTGCACA


ACCACCCCCTGCCAG GCGGATCTCCACCAT CTACTCCCTGCACAT


CCACCCCCTGCCAGG CGGATCTCCACCATG TACTCCCTGCACATC


35CACCCCCTGCCAGGA GGATCTCCACCATGC ACTCCCTGCACATCC


ACCCCCTGCCAGGAC GATCTCCACCATGCG CTCCCTGCACATCCC


CCCCCTGCCAGGACT ATCTCCACCATGCGC TCCCTGCACATCCCC


CCCCTGCCAGGACTC TCTCCACCATGCGCC CCCTGCACATCCCCA


CCCTGCCAGGACTCC CTCCACCATGCGCCT CCTGCACATCCCCAA


40CCTGCCAGGACTCCC TCCACCATGCGCCTT CTGCACATCCCCAAC


CTGCCAGGACTCCCT CCACCATGCGCCTTC TGCACATCCCCAACT


TGCCAGGACTCCCTG CACCATGCGCCTTCC GCACATCCCCAACTG


GCCAGGACTCCCTGC ACCATGCGCCTTCCG CACATCCCCAACTGT


CCAGGACTCCCTGCC CCATGCGCCTTCCGG ~ACATCCCCAACTGTG


45CAGGACTCCCTGCCA CATGCGCCTTCCGGA CATCCCCAACTGTGA


AGGACTCCCTGCCAA ATGCGCCTTCCGGAT ATCCCCAACTGTGAC


GGACTCCCTGCCAAC TGCGCCTTCCGGATG TCCCCAACTGTGACA


GACTCCCTGCCAACA GCGCCTTCCGGATGA CCCCAACTGTGACAA


ACTCCCTGCCAACAG CGCCTTCCGGATGAG CCCAACTGTGACAAG


50CTCCCTGCCAACAGG GCCTTCCGGATGAGC CCAACTGTGACAAGC


TCCCTGCCAACAGGA CCTTCCGGATGAGCG CAACTGTGACAAGCA




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AACTGTGACAAGCAT AACGGGCAGCGTGGG ATCCAGGGAGCCCCC


ACTGTGACAAGCATG ACGGGCAGCGTGGGG TCCAGGGAGCCCCCA


CTGTGACAAGCATGG CGGGCAGCGTGGGGA CCAGGGAGCCCCCAC


TGTGACAAGCATGGC GGGCAGCGTGGGGAG CAGGGAGCCCCCACC


GTGACAAGCATGGCC GGCAGCGTGGGGAGT AGGGAGCCCCCACCA


TGACAAGCATGGCCT GCAGCGTGGGGAGTG GGGAGCCCCCACCAT


GACAAGCATGGCCTG CAGCGTGGGGAGTGC GGAGCCCCCACCATC


ACAAGCATGGCCTGT AGCGTGGGGAGTGCT GAGCCCCCACCATCC


CAAGCATGGCCTGTA GCGTGGGGAGTGCTG AGCCCCCACCATCCG


10AAGCATGGCCTGTAC CGTGGGGAGTGCTGG GCCCCCACCATCCGG


AGCATGGCCTGTACA GTGGGGAGTGCTGGT CCCCCACCATCCGGG


GCATGGCCTGTACAA TGGGGAGTGCTGGTG CCCCACCATCCGGGG


CATGGCCTGTACAAC GGGGAGTGCTGGTGT CCCACCATCCGGGGG


ATGGCCTGTACAACC GGGAGTGCTGGTGTG CCACCATCCGGGGGG


15TGGCCTGTACAACCT GGAGTGCTGGTGTGT CACCATCCGGGGGGA


GGCCTGTACAACCTC GAGTGCTGGTGTGTG ACCATCCGGGGGGAC


GCCTGTACAACCTCA AGTGCTGGTGTGTGA CCATCCGGGGGGACC


CCTGTACAACCTCAA GTGCTGGTGTGTGAA CATCCGGGGGGACCC


CTGTACAACCTCAAA TGCTGGTGTGTGAAC ATCCGGGGGGACCCC


20TGTACAACCTCAAAC GCTGGTGTGTGAACC TCCGGGGGGACCCCG


GTACAACCTCAA~CA CTGGTGTGTGAACCC CCGGGGGGACCCCGA


TACAACCTCAAACAG TGGTGTGTGAACCCC CGGGGGGACCCCGAG


ACAACCTCAAACAGT GGTGTGTGAACCCCA GGGGGGACCCCGAGT


CAACCTCAAACAGTG GTGTGTGAACCCCAA GGGGGACCCCGAGTG


25AACCTCAAACAGTGC TGTGTGAACCCCAAC GGGGACCCCGAGTGT


ACCTCAAACAGTGCA GTGTGAACCCCAACA GGGACCCCGAGTGTC


CCTCAAACAGTGCAA TGTGAACCCCAACAC GGACCCCGAGTGTCA


CTCAAACAGTGCAAG GTGAACCCCAACACC GACCCCGAGTGTCAT


TCAAACAGTGCAAGA TGAACCCCAACACCG ACCCCGAGTGTCATC


30CAAACAGTGCAAGAT GAACCCCAACACCGG CCCCGAGTGTCATCT


AAACAGTGCAAGATG AACCCCAACACCGGG CCCGAGTGTCATCTC


AACAGTGCAAGATGT ACCCCAACACCGGGA CCGAGTGTCATCTCT


ACAGTGCAAGATGTC CCCCAACACCGGGAA CGAGTGTCATCTCTT


CAGTGCAAGATGTCT CCCAACACCGGGAAG GAGTGTCATCTCTTC


35AGTGCAAGATGTCTC CCAACACCGGGAAGC AGTGTCATCTCTTCT


GTGCAAGATGTCTCT CAACACCGGGAAGCT GTGTCATCTCTTCTA


TGCAAGATGTCTCTG AACACCGGGAAGCTG TGTCATCTCTTCTAC


GCAAGATGTCTCTGA ACACCGGGAAGCTGA GTCATCTCTTCTACA


CAAGATGTCTCTGAA CACCGGGAAGCTGAT TCATCTCTTCTACAA


40AAGATGTCTCTGAAC ACCGGGAAGCTGATC CATCTCTTCTACAAT


AGATGTCTCTGAACG CCGGGAAGCTGATCC ATCTCTTCTACAATG


GATGTCTCTGAACGG CGGGAAGCTGATCCA TCTCTTCTACAATGA


ATGTCTCTGAACGGG GGGAAGCTGATCCAG CTCTTCTACAATGAG


TGTCTCTGAACGGGC GGAAGCTGATCCAGG TCTTCTACAATGAGC


45GTCTCTGAACGGGCA GAAGCTGATCCAGGG CTTCTACAATGAGCA


TCTCTGAACGGGCAG AAGCTGATCCAGGGA TTCTACAATGAGCAG


CTCTGAACGGGCAGC AGCTGATCCAGGGAG TCTACAATGAGCAGC


TCTGAACGGGCAGCG GCTGATCCAGGGAGC CTACAATGAGCAGCA


CTGAACGGGCAGCGT CTGATCCAGGGAGCC TACAATGAGCAGCAG


50TGAACGGGCAGCGTG TGATCCAGGGAGCCC ACAATGAGCAGCAGG


GAACGGGCAGCGTGG GATCCAGGGAGCCCC CAATGAGCAGCAGGA




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AATGAGCAGCAGGAG GCAGCCAGCCGGTGC GCAGAAAACGGAGAG


ATGAGCAGCAGGAGG CAGCCAGCCGGTGCC CAGAAAACGGAGAGT


TGAGCAGCAGGAGGC AGCCAGCCGGTGCCT AGAAAACGGAGAGTG


GAGCAGCAGGAGGCT GCCAGCCGGTGCCTG GAAAACGGAGAGTGC


AGCAGCAGGAGGCTT CCAGCCGGTGCCTGG AAAACGGAGAGTGCT


GCAGCAGGAGGCTTG CAGCCGGTGCCTGGC AAACGGAGAGTGCTT


CAGCAGGAGGCTTGC AGCCGGTGCCTGGCG AACGGAGAGTGCTTG


AGCAGGAGGCTTGCG GCCGGTGCCTGGCGC ACGGAGAGTGCTTGG


GCAGGAGGCTTGCGG CCGGTGCCTGGCGCC CGGAGAGTGCTTGGG


10CAGGAGGCTTGCGGG CGGTGCCTGGCGCCC GGAGAGTGCTTGGGT


AGGAGGCTTGCGGGG GGTGCCTGGCGCCCC GAGAGTGCTTGGGTG


GGAGGCTTGCGGGGT GTGCCTGGCGCCCCT AGAGTGCTTGGGTGG


GAGGCTTGCGGGGTG TGCCTGGCGCCCCTG GAGTGCTTGGGTGGT


AGGCTTGCGGGGTGC GCCTGGCGCCCCTGC AGTGCTTGGGTGGTG


15GGCTTGCGGGGTGCA CCTGGCGCCCCTGCC GTGCTTGGGTGGTGG


GCTTGCGGGGTGCAC CTGGCGCCCCTGCCC TGCTTGGGTGGTGGG


CTTGCGGGGTGCACA TGGCGCCCCTGCCCC GCTTGGGTGGTGGGT


TTGCGGGGTGCACAC GGCGCCCCTGCCCCC CTTGGGTGGTGGGTG


TGCGGGGTGCACACC GCGCCCCTGCCCCCC TTGGGTGGTGGGTGC


20GCGGGGTGCACACCC CGCCCCTGCCCCCCG TGGGTGGTGGGTGCT


CGGGGTGCACACCCA GCCCCTGCCCCCCGC GGGTGGTGGGTGCTG


GGGGTGCACACCCAG CCCCTGCCCCCCGCC GGTGGTGGGTGCTGG


GGGTGCACACCCAGC CCCTGCCCCCCGCCC GTGGTGGGTGCTGGA


GGTGCACACCCAGCG CCTGCCCCCCGCCCC TGGTGGGTGCTGGAG


25GTGCACACCCAGCGG CTGCCCCCCGCCCCT GGTGGGTGCTGGAGG


TGCACACCCAGCGGA TGCCCCCCGCCCCTC GTGGGTGCTGGAGGA


GCACACCCAGCGGAT GCCCCCCGCCCCTCT TGGGTGCTGGAGGAT


CACACCCAGCGGATG CCCCCCGCCCCTCTC GGGTGCTGGAGGATT


ACACCCAGCGGATGC CCCCCGCCCCTCTCC GGTGCTGGAGGATTT


30CACCCAGCGGATGCA CCCCGCCCCTCTCCA GTGCTGGAGGATTTT


ACCCAGCGGATGCAG CCCGCCCCTCTCCAA TGCTGGAGGATTTTC


CCCAGCGGATGCAGT CCGCCCCTCTCCAAA GCTGGAGGATTTTCC


CCAGCGGATGCAGTA CGCCCCTCTCCAAAC CTGGAGGATTTTCCA


CAGCGGATGCAGTAG GCCCCTCTCCAAACA TGGAGGATTTTCCAG


35AGCGGATGCAGTAGA CCCCTCTCCAAACAC GGAGGATTTTCCAGT


GCGGATGCAGTAGAC CCCTCTCCAAACACC GAGGATTTTCCAGTT


CGGATGCAGTAGACC CCTCTCCAAACACCG AGGATTTTCCAGTTC


GGATGCAGTAGACCG CTCTCCAAACACCGG GGATTTTCCAGTTCT


GATGCAGTAGACCGC TCTCCAAACACCGGC GATTTTCCAGTTCTG


40ATGCAGTAGACCGCA CTCCAAACACCGGCA ATTTTCCAGTTCTGA


TGCAGTAGACCGCAG TCCAAACACCGGCAG TTTTCCAGTTCTGAC


GCAGTAGACCGCAGC CCAAACACCGGCAGA TTTCCAGTTCTGACA


CAGTAGACCGCAGCC CAAACACCGGCAGAA TTCCAGTTCTGACAC


AGTAGACCGCAGCCA AAACACCGGCAGAAA TCCAGTTCTGACACA


45GTAGACCGCAGCCAG AACACCGGCAGAAAA CCAGTTCTGACACAC


TAGACCGCAGCCAGC ACACCGGCAGAAAAC CAGTTCTGACACACG


AGACCGCAGCCAGCC CACCGGCAGAAAACG AGTTCTGACACACGT


GACCGCAGCCAGCCG ACCGGCAGAAAACGG GTTCTGACACACGTA


ACCGCAGCCAGCCGG CCGGCAGAAAACGGA TTCTGACACACGTAT


50CCGCAGCCAGCCGGT CGGCAGAAAACGGAG TCTGACACACGTATT


CGCAGCCAGCCGGTG GGCAGAAAACGGAGA CTGACACACGTATTT




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TGACACACGTATTTA CCCGGCCTCTCTCTT TCCCCGGGGGAGGAA


GACACACGTATTTAT CCGGCCTCTCTCTTC CCCCGGGGGAGGAAG


ACACACGTATTTATA CGGCCTCTCTCTTCC CCCGGGGGAGGAAGG


CACACGTATTTATAT GGCCTCTCTCTTCCC CCGGGGGAGGAAGGG


ACACGTATTTATATT GCCTCTCTCTTCCCA CGGGGGAGGAAGGGG


CACGTATTTATATTT CCTCTCTCTTCCCAG GGGGGAGGAAGGGGG


ACGTATTTATATTTG CTCTCTCTTCCCAGC GGGGAGGAAGGGGGT


CGTATTTATATTTGG TCTCTCTTCCCAGCT GGGAGGAAGGGGGTT


GTATTTATATTTGGA CTCTCTTCCCAGCTG GGAGGAAGGGGGTTG


10TATTTATATTTGGAA TCTCTTCCCAGCTGC GAGGAAGGGGGTTGT


ATTTATATTTGGAAA CTCTTCCCAGCTGCA AGGAAGGGGGTTGTG


TTTATATTTGGAAAG TCTTCCCAGCTGCAG GGAAGGGGGTTGTGG


TTATATTTGGAAAGA CTTCCCAGCTGCAGA GAAGGGGGTTGTGGT


TATATTTGGAAAGAG TTCCCAGCTGCAGAT AAGGGGGTTGTGGTC


15ATATTTGGAAAGAGA TCCCAGCTGCAGATG AGGGGGTTGTGGTCG


TATTTGGAAAGAGAC CCCAGCTGCAGATGC GGGGGTTGTGGTCGG


ATTTGGAAAGAGACC CCAGCTGCAGATGCC GGGGTTGTGGTCGGG


TTTGGAAAGAGACCA CAGCTGCAGATGCCA GGGTTGTGGTCGGGG


TTGGAAAGAGACCAG AGCTGCAGATGCCAC GGTTGTGGTCGGGGA


20TGGAAAGAGACCAGC GCTGCAGATGCCACA GTTGTGGTCGGGGAG


GGAAAGAGACCAGCA CTGCAGATGCCACAC TTGTGGTCGGGGAGC


GAAAGAGACCAGCAC TGCAGATGCCACACC TGTGGTCGGGGAGCT


AAAGAGACCAGCACC GCAGATGCCACACCT GTGGTCGGGGAGCTG


AAGAGACCAGCACCG CAGATGCCACACCTG TGGTCGGGGAGCTGG


25AGAGACCAGCACCGA AGATGCCACACCTGC GGTCGGGGAGCTGGG


GAGACCAGCACCGAG GATGCCACACCTGCT GTCGGGGAGCTGGGG


AGACCAGCACCGAGC ATGCCACACCTGCTC TCGGGGAGCTGGGGT


GACCAGCACCGAGCT TGCCACACCTGCTCC CGGGGAGCTGGGGTA


ACCAGCACCGAGCTC GCCACACCTGCTCCT GGGGAGCTGGGGTAC


30CCAGCACCGAGCTCG CCACACCTGCTCCTT GGGAGCTGGGGTACA


CAGCACCGAGCTCGG CACACCTGCTCCTTC GGAGCTGGGGTACAG


AGCACCGAGCTCGGC ACACCTGCTCCTTCT GAGCTGGGGTACAGG


GCACCGAGCTCGGCA CACCTGCTCCTTCTT AGCTGGGGTACAGGT


CACCGAGCTCGGCAC ACCTGCTCCTTCTTG GCTGGGGTACAGGTT


35ACCGAGCTCGGCACC CCTGCTCCTTCTTGC CTGGGGTACAGGTTT


CCGAGCTCGGCACCT CTGCTCCTTCTTGCT TGGGGTACAGGTTTG


CGAGCTCGGCACCTC TGCTCCTTCTTGCTT GGGGTACAGGTTTGG


GAGCTCGGCACCTCC GCTCCTTCTTGCTTT GGGTACAGGTTTGGG


AGCTCGGCACCTCCC CTCCTTCTTGCTTTC GGTACAGGTTTGGGG


40GCTCGGCACCTCCCC TCCTTCTTGCTTTCC GTACAGGTTTGGGGA


CTCGGCACCTCCCCG CCTTCTTGCTTTCCC TACAGGTTTGGGGAG


TCGGCACCTCCCCGG CTTCTTGCTTTCCCC ACAGGTTTGGGGAGG


CGGCACCTCCCCGGC TTCTTGCTTTCCCCG CAGGTTTGGGGAGGG


GGCACCTCCCCGGCC TCTTGCTTTCCCCGG AGGTTTGGGGAGGGG


45GCACCTCCCCGGCCT CTTGCTTTCCCCGGG GGTTTGGGGAGGGGG


CACCTCCCCGGCCTC TTGCTTTCCCCGGGG GTTTGGGGAGGGGGA


ACCTCCCCGGCCTCT TGCTTTCCCCGGGGG TTTGGGGAGGGGGAA


CCTCCCCGGCCTCTC GCTTTCCCCGGGGGA TTGGGGAGGGGGAAG


CTCCCCGGCCTCTCT CTTTCCCCGGGGGAG TGGGGAGGGGGAAGA


50TCCCCGGCCTCTCTC TTTCCCCGGGGGAGG GGGGAGGGGGAAGAG


CCCCGGCCTCTCTCT TTCCCCGGGGGAGGA GGGAGGGGGAAGAGA




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
- 43 -
GGAGGGGGAAGAGAA AGATTAAAGGAAGGA


GAGGGGGAAGAGAAA GATTAAAGGAAGGAA


AGGGGGAAGAGAAAT ATTAAAGGAAGGAAA


GGGGGAAGAGAAATT TTAAAGGAAGGAAAA


GGGGAAGAGAAATTT TAAAGGAAGGAAAAG


GGGAAGAGAAATTTT AAAGGAAGGAAAAGT


GGAAGAGAAATTTTT


GAAGAGAAATTTTTA


AAGAGAAATTTTTAT


10AGAGAAATTTTTATT


GAGAAATTTTTATTT


AGAAATTTTTATTTT


GAAATTTTTATTTTT


AAATTTTTATTTTTG


15AATTTTTATTTTTGA


ATTTTTATTTTTGAA


TTTTTATTTTTGAAC


TTTTATTTTTGAACC


TTTATTTTTGAACCC


20TTATTTTTGAACCCC


TATTTTTGAACCCCT


ATTTTTGAACCCCTG


TTTTTGAACCCCTGT


TTTTGAACCCCTGTG


25TTTGAACCCCTGTGT


TTGAACCCCTGTGTC


TGAACCCCTGTGTCC


GAACCCCTGTGTCCC


AACCCCTGTGTCCCT


30ACCCCTGTGTCCCTT


CCCCTGTGTCCCTTT


CCCTGTGTCCCTTTT


CCTGTGTCCCTTTTG


CTGTGTCCCTTTTGC


35TGTGTCCCTTTTGCA


GTGTCCCTTTTGCAT


TGTCCCTTTTGCATA


GTCCCTTTTGCATAA


TCCCTTTTGCATAAG


40CCCTTTTGCATAAGA


CCTTTTGCATAAGAT


CTTTTGCATAAGATT


TTTTGCATAAGATTA


TTTGCATAAGATTAA


45TTGCATAAGATTAAA


TGCATAAGATTAAAG


GCATAAGATTAAAGG


CATAAGATTAAAGGA


ATAAGATTAAAGGAA


50TAAGATTAAAGGAAG


AAGATTAAAGGAAGG




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-44-
EXAMPLE 7
Antisense oligonucleotides to IGFBP3 may be selected from molecules capable of
interacting
S with one or more of the following sense oligonucleotides:
CTCAGCGCCCAGCCG GCCGTGTACTGTCGC GCAGCGTGCCCCGGT


TCAGCGCCCAGCCGC CCGTGTACTGTCGCC CAGCGTGCCCCGGTT


CAGCGCCCAGCCGCT CGTGTACTGTCGCCC AGCGTGCCCCGGTTG


10AGCGCCCAGCCGCTT GTGTACTGTCGCCCC GCGTGCCCCGGTTGC


GCGCCCAGCCGCTTC TGTACTGTCGCCCCA CGTGCCCCGGTTGCA


CGCCCAGCCGCTTCC GTACTGTCGCCCCAT GTGCCCCGGTTGCAG


GCCCAGCCGCTTCCT TACTGTCGCCCCATC TGCCCCGGTTGCAGG


CCCAGCCGCTTCCTG ACTGTCGCCCCATCC GCCCCGGTTGCAGGC


15CCAGCCGCTTCCTGC CTGTCGCCCCATCCC CCCCGGTTGCAGGCG


CAGCCGCTTCCTGCC TGTCGCCCCATCCCT CCCGGTTGCAGGCGT


AGCCGCTTCCTGCCT GTCGCCCCATCCCTG CCGGTTGCAGGCGTC


GCCGCTTCCTGCCTG TCGCCCCATCCCTGC CGGTTGCAGGCGTCA


CCGCTTCCTGCCTGG CGCCCCATCCCTGCG GGTTGCAGGCGTCAT


20CGCTTCCTGCCTGGA GCCCCATCCCTGCGC GTTGCAGGCGTCATG


GCTTCCTGCCTGGAT CCCCATCCCTGCGCG TTGCAGGCGTCATGC


CTTCCTGCCTGGATT CCCATCCCTGCGCGC TGCAGGCGTCATGCA


TTCCTGCCTGGATTC CCATCCCTGCGCGCC GCAGGCGTCATGCAG


TCCTGCCTGGATTCC CATCCCTGCGCGCCC CAGGCGTCATGCAGC


25CCTGCCTGGATTCCA ATCCCTGCGCGCCCA AGGCGTCATGCAGCG


CTGCCTGGATTCCAC TCCCTGCGCGCCCAG GGCGTCATGCAGCGG


TGCCTGGATTCCACA CCCTGCGCGCCCAGC GCGTCATGCAGCGGG


GCCTGGATTCCACAG CCTGCGCGCCCAGCC CGTCATGCAGCGGGC


CCTGGATTCCACAGC CTGCGCGCCCAGCCT GTCATGCAGCGGGCG


30CTGGATTCCACAGCT TGCGCGCCCAGCCTG TCATGCAGCGGGCGC


TGGATTCCACAGCTT GCGCGCCCAGCCTGC CATGCAGCGGGCGCG


GGATTCCACAGCTTC CGCGCCCAGCCTGCC ATGCAGCGGGCGCGA


GATTCCACAGCTTCG GCGCCCAGCCTGCCA TGCAGCGGGCGCGAC


ATTCCACAGCTTCGC CGCCCAGCCTGCCAA GCAGCGGGCGCGACC


35TTCCACAGCTTCGCG GCCCAGCCTGCCAAG CAGCGGGCGCGACCC


TCCACAGCTTCGCGC CCCAGCCTGCCAAGC AGCGGGCGCGACCCA


CCACAGCTTCGCGCC CCAGCCTGCCAAGCA GCGGGCGCGACCCAC


CACAGCTTCGCGCCG CAGCCTGCCAAGCAG CGGGCGCGACCCACG


ACAGCTTCGCGCCGT AGCCTGCCAAGCAGC GGGCGCGACCCACGC


40CAGCTTCGCGCCGTG GCCTGCCAAGCAGCG GGCGCGACCCACGCT


AGCTTCGCGCCGTGT CCTGCCAAGCAGCGT GCGCGACCCACGCTC


GCTTCGCGCCGTGTA CTGCCAAGCAGCGTG CGCGACCCACGCTCT


CTTCGCGCCGTGTAC TGCCAAGCAGCGTGC GCGACCCACGCTCTG


TTCGCGCCGTGTACT GCCAAGCAGCGTGCC CGACCCACGCTCTGG


45TCGCGCCGTGTACTG CCAAGCAGCGTGCCC GACCCACGCTCTGGG


CGCGCCGTGTACTGT CAAGCAGCGTGCCCC ACCCACGCTCTGGGC


GCGCCGTGTACTGTC AAGCAGCGTGCCCCG CCCACGCTCTGGGCC


CGCCGTGTACTGTCG AGCAGCGTGCCCCGG CCACGCTCTGGGCCG




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-45-
CACGCTCTGGGCCGC GGTGGCGCGGGCTGG CGAGCCGTGCGACGC


ACGCTCTGGGCCGCT GTGGCGCGGGCTGGC GAGCCGTGCGACGCG


CGCTCTGGGCCGCTG TGGCGCGGGCTGGCG AGCCGTGCGACGCGC


GCTCTGGGCCGCTGC GGCGCGGGCTGGCGC GCCGTGCGACGCGCG


CTCTGGGCCGCTGCG GCGCGGGCTGGCGCG CCGTGCGACGCGCGT


TCTGGGCCGCTGCGC CGCGGGCTGGCGCGA CGTGCGACGCGCGTG


CTGGGCCGCTGCGCT GCGGGCTGGCGCGAG GTGCGACGCGCGTGC


TGGGCCGCTGCGCTG CGGGCTGGCGCGAGC TGCGACGCGCGTGCA


GGGCCGCTGCGCTGA GGGCTGGCGCGAGCT GCGACGCGCGTGCAC


10GGCCGCTGCGCTGAC GGCTGGCGCGAGCTC CGACGCGCGTGCACT


GCCGCTGCGCTGACT GCTGGCGCGAGCTCG GACGCGCGTGCACTG


CCGCTGCGCTGACTC CTGGCGCGAGCTCGG ACGCGCGTGCACTGG


CGCTGCGCTGACTCT TGGCGCGAGCTCGGG CGCGCGTGCACTGGC


GCTGCGCTGACTCTG GGCGCGAGCTCGGGG GCGCGTGCACTGGCC


15CTGCGCTGACTCTGC GCGCGAGCTCGGGGG CGCGTGCACTGGCCC


TGCGCTGACTCTGCT CGCGAGCTCGGGGGG GCGTGCACTGGCCCA


GCGCTGACTCTGCTG GCGAGCTCGGGGGGC CGTGCACTGGCCCAG


CGCTGACTCTGCTGG CGAGCTCGGGGGGCT GTGCACTGGCCCAGT


GCTGACTCTGCTGGT GAGCTCGGGGGGCTT TGCACTGGCCCAGTG


20CTGACTCTGCTGGTG AGCTCGGGGGGCTTG GCACTGGCCCAGTGC


TGACTCTGCTGGTGC GCTCGGGGGGCTTGG CACTGGCCCAGTGCG


GACTCTGCTGGTGCT CTCGGGGGGCTTGGG ACTGGCCCAGTGCGC


ACTCTGCTGGTGCTG TCGGGGGGCTTGGGT CTGGCCCAGTGCGCG


CTCTGCTGGTGCTGC CGGGGGGCTTGGGTC TGGCCCAGTGCGCGC


25TCTGCTGGTGCTGCT GGGGGGCTTGGGTCC GGCCCAGTGCGCGCC


CTGCTGGTGCTGCTC GGGGGCTTGGGTCCC GCCCAGTGCGCGCCT


TGCTGGTGCTGCTCC GGGGCTTGGGTCCCG CCCAGTGCGCGCCTC


GCTGGTGCTGCTCCG GGGCTTGGGTCCCGT CCAGTGCGCGCCTCC


CTGGTGCTGCTCCGC GGCTTGGGTCCCGTG CAGTGCGCGCCTCCG


30TGGTGCTGCTCCGCG GCTTGGGTCCCGTGG AGTGCGCGCCTCCGC


GGTGCTGCTCCGCGG CTTGGGTCCCGTGGT GTGCGCGCCTCCGCC


GTGCTGCTCCGCGGG TTGGGTCCCGTGGTG TGCGCGCCTCCGCCC


TGCTGCTCCGCGGGC TGGGTCCCGTGGTGC GCGCGCCTCCGCCCG


GCTGCTCCGCGGGCC GGGTCCCGTGGTGCG CGCGCCTCCGCCCGC


35CTGCTCCGCGGGCCG GGTCCCGTGGTGCGC GCGCCTCCGCCCGCC


TGCTCCGCGGGCCGC GTCCCGTGGTGCGCT CGCCTCCGCCCGCCG


GCTCCGCGGGCCGCC TCCCGTGGTGCGCTG GCCTCCGCCCGCCGT


CTCCGCGGGCCGCCG CCCGTGGTGCGCTGC CCTCCGCCCGCCGTG


TCCGCGGGCCGCCGG CCGTGGTGCGCTGCG CTCCGCCCGCCGTGT


40CCGCGGGCCGCCGGT CGTGGTGCGCTGCGA TCCGCCCGCCGTGTG


CGCGGGCCGCCGGTG GTGGTGCGCTGCGAG CCGCCCGCCGTGTGC


GCGGGCCGCCGGTGG TGGTGCGCTGCGAGC CGCCCGCCGTGTGCG


CGGGCCGCCGGTGGC GGTGCGCTGCGAGCC GCCCGCCGTGTGCGC


GGGCCGCCGGTGGCG GTGCGCTGCGAGCCG CCCGCCGTGTGCGCG


45GGCCGCCGGTGGCGC TGCGCTGCGAGCCGT CCGCCGTGTGCGCGG


GCCGCCGGTGGCGCG GCGCTGCGAGCCGTG CGCCGTGTGCGCGGA


CCGCCGGTGGCGCGG CGCTGCGAGCCGTGC GCCGTGTGCGCGGAG


CGCCGGTGGCGCGGG GCTGCGAGCCGTGCG CCGTGTGCGCGGAGC


GCCGGTGGCGCGGGC CTGCGAGCCGTGCGA CGTGTGCGCGGAGCT


50CCGGTGGCGCGGGCT TGCGAGCCGTGCGAC GTGTGCGCGGAGCTG


CGGTGGCGCGGGCTG GCGAGCCGTGCGACG TGTGCGCGGAGCTGG




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-46-
GTGCGCGGAGCTGGT ACTGAGCGAGGGCCA CCTTCGCTGCCAGCC


TGCGCGGAGCTGGTG CTGAGCGAGGGCCAG CTTCGCTGCCAGCCG


GCGCGGAGCTGGTGC TGAGCGAGGGCCAGC TTCGCTGCCAGCCGT


CGCGGAGCTGGTGCG GAGCGAGGGCCAGCC TCGCTGCCAGCCGTC


GCGGAGCTGGTGCGC AGCGAGGGCCAGCCG CGCTGCCAGCCGTCG


CGGAGCTGGTGCGCG GCGAGGGCCAGCCGT GCTGCCAGCCGTCGC


GGAGCTGGTGCGCGA CGAGGGCCAGCCGTG CTGCCAGCCGTCGCC


GAGCTGGTGCGCGAG GAGGGCCAGCCGTGC TGCCAGCCGTCGCCC


AGCTGGTGCGCGAGC AGGGCCAGCCGTGCG GCCAGCCGTCGCCCG


10GCTGGTGCGCGAGCC GGGCCAGCCGTGCGG CCAGCCGTCGCCCGA


CTGGTGCGCGAGCCG GGCCAGCCGTGCGGC CAGCCGTCGCCCGAC


TGGTGCGCGAGCCGG GCCAGCCGTGCGGCA AGCCGTCGCCCGACG


GGTGCGCGAGCCGGG CCAGCCGTGCGGCAT GCCGTCGCCCGACGA


GTGCGCGAGCCGGGC CAGCCGTGCGGCATC CCGTCGCCCGACGAG


15TGCGCGAGCCGGGCT AGCCGTGCGGCATCT CGTCGCCCGACGAGG


GCGCGAGCCGGGCTG GCCGTGCGGCATCTA GTCGCCCGACGAGGC


CGCGAGCCGGGCTGC CCGTGCGGCATCTAC TCGCCCGACGAGGCG


GCGAGCCGGGCTGCG ' CGTGCGGCATCTACA CGCCCGACGAGGCGC


CGAGCCGGGCTGCGG GTGCGGCATCTACAC. GCCCGACGAGGCGCG


20GAGCCGGGCTGCGGC TGCGGCATCTACACC CCCGACGAGGCGCGA


AGCCGGGCTGCGGCT GCGGCATCTACACCG CCGACGAGGCGCGAC


GCCGGGCTGCGGCTG CGGCATCTACACCGA CGACGAGGCGCGACC


CCGGGCTGCGGCTGC GGCATCTACACCGAG GACGAGGCGCGACCG


CGGGCTGCGGCTGCT GCATCTACACCGAGC ACGAGGCGCGACCGC


25GGGCTGCGGCTGCTG CATCTACACCGAGCG CGAGGCGCGACCGCT


GGCTGCGGCTGCTGC ATCTACACCGAGCGC GAGGCGCGACCGCTG


GCTGCGGCTGCTGCC TCTACACCGAGCGCT AGGCGCGACCGCTGC


CTGCGGCTGCTGCCT CTACACCGAGCGCTG GGCGCGACCGCTGCA


TGCGGCTGCTGCCTG TACACCGAGCGCTGT GCGCGACCGCTGCAG


30GCGGCTGCTGCCTGA ACACCGAGCGCTGTG CGCGACCGCTGCAGG


CGGCTGCTGCCTGAC CACCGAGCGCTGTGG GCGACCGCTGCAGGC


GGCTGCTGCCTGACG ACCGAGCGCTGTGGC CGACCGCTGCAGGCG


GCTGCTGCCTGACGT CCGAGCGCTGTGGCT GACCGCTGCAGGCGC


CTGCTGCCTGACGTG CGAGCGCTGTGGCTC ACCGCTGCAGGCGCT


35TGCTGCCTGACGTGC GAGCGCTGTGGCTCC CCGCTGCAGGCGCTG


GCTGCCTGACGTGCG AGCGCTGTGGCTCCG CGCTGCAGGCGCTGC


CTGCCTGACGTGCGC GCGCTGTGGCTCCGG GCTGCAGGCGCTGCT


TGCCTGACGTGCGCA CGCTGTGGCTCCGGC CTGCAGGCGCTGCTG


GCCTGACGTGCGCAC GCTGTGGCTCCGGCC TGCAGGCGCTGCTGG


40CCTGACGTGCGCACT CTGTGGCTCCGGCCT GCAGGCGCTGCTGGA


CTGACGTGCGCACTG TGTGGCTCCGGCCTT CAGGCGCTGCTGGAC


TGACGTGCGCACTGA GTGGCTCCGGCCTTC AGGCGCTGCTGGACG


GACGTGCGCACTGAG TGGCTCCGGCCTTCG GGCGCTGCTGGACGG


ACGTGCGCACTGAGC GGCTCCGGCCTTCGC GCGCTGCTGGACGGC


45CGTGCGCACTGAGCG GCTCCGGCCTTCGCT CGCTGCTGGACGGCC


GTGCGCACTGAGCGA CTCCGGCCTTCGCTG GCTGCTGGACGGCCG


TGCGCACTGAGCGAG TCCGGCCTTCGCTGC CTGCTGGACGGCCGC


GCGCACTGAGCGAGG CCGGCCTTCGCTGCC TGCTGGACGGCCGCG


CGCACTGAGCGAGGG CGGCCTTCGCTGCCA GCTGGACGGCCGCGG


SOGCACTGAGCGAGGGC GGCCTTCGCTGCCAG CTGGACGGCCGCGGG


CACTGAGCGAGGGCC GCCTTCGCTGCCAGC TGGACGGCCGCGGGC




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-47-
GGACGGCCGCGGGCT CTACCTGCTGCCAGC AGACCGCAGCGCCGG


GACGGCCGCGGGCTC TACCTGCTGCCAGCG GACCGCAGCGCCGGC


ACGGCCGCGGGCTCT ACCTGCTGCCAGCGC ACCGCAGCGCCGGCA


CGGCCGCGGGCTCTG CCTGCTGCCAGCGCC CCGCAGCGCCGGCAG


GGCCGCGGGCTCTGC CTGCTGCCAGCGCCG CGCAGCGCCGGCAGT


GCCGCGGGCTCTGCG TGCTGCCAGCGCCGC GCAGCGCCGGCAGTG


CCGCGGGCTCTGCGT GCTGCCAGCGCCGCC CAGCGCCGGCAGTGT


CGCGGGCTCTGCGTC CTGCCAGCGCCGCCA AGCGCCGGCAGTGTG


GCGGGCTCTGCGTCA TGCCAGCGCCGCCAG GCGCCGGCAGTGTGG


10CGGGCTCTGCGTCAA GCCAGCGCCGCCAGC CGCCGGCAGTGTGGA


GGGCTCTGCGTCAAC CCAGCGCCGCCAGCT GCCGGCAGTGTGGAG


GGCTCTGCGTCAACG CAGCGCCGCCAGCTC CCGGCAGTGTGGAGA


GCTCTGCGTCAACGC AGCGCCGCCAGCTCC CGGCAGTGTGGAGAG


CTCTGCGTCAACGCT GCGCCGCCAGCTCCA GGCAGTGTGGAGAGC


15TCTGCGTCAACGCTA CGCCGCCAGCTCCAG GCAGTGTGGAGAGCC


CTGCGTCAACGCTAG GCCGCCAGCTCCAGG CAGTGTGGAGAGCCC


TGCGTCAACGCTAGT CCGCCAGCTCCAGGA AGTGTGGAGAGCCCG


GCGTCAACGCTAGTG CGCCAGCTCCAGGAA GTGTGGAGAGCCCGT


CGTCAACGCTAGTGC GCCAGCTCCAGGAAA TGTGGAGAGCCCGTC


20GTCAACGCTAGTGCC CCAGCTCCAGGAAAT GTGGAGAGCCCGTCC


TCAACGCTAGTGCCG CAGCTCCAGGAAATG TGGAGAGCCCGTCCG


CAACGCTAGTGCCGT AGCTCCAGGAAATGC GGAGAGCCCGTCCGT


AACGCTAGTGCCGTC GCTCCAGGAAATGCT GAGAGCCCGTCCGTC


ACGCTAGTGCCGTCA CTCCAGGAAATGCTA AGAGCCCGTCCGTCT


25CGCTAGTGCCGTCAG TCCAGGAAATGCTAG GAGCCCGTCCGTCTC


GCTAGTGCCGTCAGC CCAGGAAATGCTAGT AGCCCGTCCGTCTCC


CTAGTGCCGTCAGCC CAGGAAATGCTAGTG GCCCGTCCGTCTCCA


TAGTGCCGTCAGCCG AGGAAATGCTAGTGA CCCGTCCGTCTCCAG


AGTGCCGTCAGCCGC GGAAATGCTAGTGAG CCGTCCGTCTCCAGC


30GTGCCGTCAGCCGCC GAAATGCTAGTGAGT CGTCCGTCTCCAGCA


TGCCGTCAGCCGCCT AAATGCTAGTGAGTC GTCCGTCTCCAGCAC


GCCGTCAGCCGCCTG AATGCTAGTGAGTCG TCCGTCTCCAGCACG


CCGTCAGCCGCCTGC ATGCTAGTGAGTCGG CCGTCTCCAGCACGC


CGTCAGCCGCCTGCG TGCTAGTGAGTCGGA CGTCTCCAGCACGCA


35GTCAGCCGCCTGCGC GCTAGTGAGTCGGAG GTCTCCAGCACGCAC


TCAGCCGCCTGCGCG CTAGTGAGTCGGAGG TCTCCAGCACGCACC


CAGCCGCCTGCGCGC TAGTGAGTCGGAGGA CTCCAGCACGCACCG


AGCCGCCTGCGCGCC AGTGAGTCGGAGGAA TCCAGCACGCACCGG


GCCGCCTGCGCGCCT GTGAGTCGGAGGAAG CCAGCACGCACCGGG


40CCGCCTGCGCGCCTA TGAGTCGGAGGAAGA CAGCACGCACCGGGT


CGCCTGCGCGCCTAC GAGTCGGAGGAAGAC AGCACGCACCGGGTG


GCCTGCGCGCCTACC AGTCGGAGGAAGACC GCACGCACCGGGTGT


CCTGCGCGCCTACCT GTCGGAGGAAGACCG CACGCACCGGGTGTC


CTGCGCGCCTACCTG TCGGAGGAAGACCGC ACGCACCGGGTGTCT


45TGCGCGCCTACCTGC CGGAGGAAGACCGCA CGCACCGGGTGTCTG


GCGCGCCTACCTGCT GGAGGAAGACCGCAG GCACCGGGTGTCTGA


CGCGCCTACCTGCTG GAGGAAGACCGCAGC CACCGGGTGTCTGAT


GCGCCTACCTGCTGC AGGAAGACCGCAGCG ACCGGGTGTCTGATC


CGCCTACCTGCTGCC GGAAGACCGCAGCGC CCGGGTGTCTGATCC


50GCCTACCTGCTGCCA GAAGACCGCAGCGCC CGGGTGTCTGATCCC


CCTACCTGCTGCCAG AAGACCGCAGCGCCG GGGTGTCTGATCCCA




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
- 48 -
GGTGTCTGATCCCAA GAAAGGGCATGCTAA GAGCACAGATACCCA


GTGTCTGATCCCAAG AAAGGGCATGCTAAA AGCACAGATACCCAG


TGTCTGATCCCAAGT AAGGGCATGCTAAAG GCACAGATACCCAGA


GTCTGATCCCAAGTT AGGGCATGCTAAAGA CACAGATACCCAGAA


TCTGATCCCAAGTTC GGGCATGCTAAAGAC ACAGATACCCAGAAC


CTGATCCCAAGTTCC GGCATGCTAAAGACA CAGATACCCAGAACT


TGATCCCAAGTTCCA GCATGCTAAAGACAG AGATACCCAGAACTT


GATCCCAAGTTCCAC CATGCTAAAGACAGC GATACCCAGAACTTC


ATCCCAAGTTCCACC ATGCTAAAGACAGCC ATACCCAGAACTTCT


10TCCCAAGTTCCACCC TGCTAAAGACAGCCA TACCCAGAACTTCTC


CCCAAGTTCCACCCC GCTAAAGACAGCCAG ACCCAGAACTTCTCC


CCAAGTTCCACCCCC CTAAAGACAGCCAGC CCCAGAACTTCTCCT


CAAGTTCCACCCCCT TAAAGACAGCCAGCG CCAGAACTTCTCCTC


AAGTTCCACCCCCTC AAAGACAGCCAGCGC CAGAACTTCTCCTCC


15AGTTCCACCCCCTCC AAGACAGCCAGCGCT AGAACTTCTCCTCCG


GTTCCACCCCCTCCA AGACAGCCAGCGCTA GAACTTCTCCTCCGA


TTCCACCCCCTCCAT GACAGCCAGCGCTAC AACTTCTCCTCCGAG


TCCACCCCCTCCATT ACAGCCAGCGCTACA ACTTCTCCTCCGAGT


CCACCCCCTCCATTC CAGCCAGCGCTACAA CTTCTCCTCCGAGTC


20CACCCCCTCCATTCA AGCCAGCGCTACAAA TTCTCCTCCGAGTCC


ACCCCCTCCATTCAA GCCAGCGCTACAAAG TCTCCTCCGAGTCCA


CCCCCTCCATTCAAA CCAGCGCTACAAAGT CTCCTCCGAGTCCAA


CCCCTCCATTCAAAG CAGCGCTACAAAGTT TCCTCCGAGTCCAAG


CCCTCCATTCAAAGA AGCGCTACAAAGTTG CCTCCGAGTCCAAGC


25CCTCCATTCAAAGAT GCGCTACAAAGTTGA CTCCGAGTCCAAGCG


CTCCATTCAAAGATA CGCTACAAAGTTGAC TCCGAGTCCAAGCGG


TCCATTCAAAGATAA GCTACAAAGTTGACT CCGAGTCCAAGCGGG


CCATTCAAAGATAAT CTACAAAGTTGACTA CGAGTCCAAGCGGGA


CATTCAAAGATAATC TACAAAGTTGACTAC GAGTCCAAGCGGGAG


30ATTCAAAGATAATCA ACAAAGTTGACTACG AGTCCAAGCGGGAGA


TTCAAAGATAATCAT CAAAGTTGACTACGA GTCCAAGCGGGAGAC


TCAAAGATAATCATC AAAGTTGACTACGAG TCCAAGCGGGAGACA


CAAAGATAATCATCA AAGTTGACTACGAGT CCAAGCGGGAGACAG


AAAGATAATCATCAT AGTTGACTACGAGTC CAAGCGGGAGACAGA


35AAGATAATCATCATC GTTGACTACGAGTCT AAGCGGGAGACAGAA


AGATAATCATCATCA TTGACTACGAGTCTC AGCGGGAGACAGAAT


GATAATCATCATCAA TGACTACGAGTCTCA GCGGGAGACAGAATA


ATAATCATCATCAAG GACTACGAGTCTCAG CGGGAGACAGAATAT


TAATCATCATCAAGA ACTACGAGTCTCAGA GGGAGACAGAATATG


40AATCATCATCAAGAA CTACGAGTCTCAGAG GGAGACAGAATATGG


ATCATCATCAAGAAA TACGAGTCTCAGAGC GAGACAGAATATGGT


TCATCATCAAGAAAG ACGAGTCTCAGAGCA AGACAGAATATGGTC


CATCATCAAGAAAGG CGAGTCTCAGAGCAC GACAGAATATGGTCC


ATCATCAAGAAAGGG GAGTCTCAGAGCACA ACAGAATATGGTCCC


45TCATCAAGAAAGGGC AGTCTCAGAGCACAG CAGAATATGGTCCCT


CATCAAGAAAGGGCA GTCTCAGAGCACAGA AGAATATGGTCCCTG


ATCAAGAAAGGGCAT TCTCAGAGCACAGAT GAATATGGTCCCTGC


TCAAGAAAGGGCATG CTCAGAGCACAGATA AATATGGTCCCTGCC


CAAGAAAGGGCATGC TCAGAGCACAGATAC ATATGGTCCCTGCCG


50AAGAAAGGGCATGCT CAGAGCACAGATACC TATGGTCCCTGCCGT


AGAAAGGGCATGCTA AGAGCACAGATACCC ATGGTCCCTGCCGTA




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-49-
TGGTCCCTGCCGTAG CAATGTGCTGAGTCC ATTTTATAAGAA.AAA


GGTCCCTGCCGTAGA AATGTGCTGAGTCCC TTTTATAAGAAAAAG


GTCCCTGCCGTAGAG ATGTGCTGAGTCCCA TTTATAAGAAAAAGC


TCCCTGCCGTAGAGA TGTGCTGAGTCCCAG TTATAAGAAAAAGCA


CCCTGCCGTAGAGAA GTGCTGAGTCCCAGG TATAAGAAAAAGCAG


CCTGCCGTAGAGAAA TGCTGAGTCCCAGGG ATAAGAAAAAGCAGT


CTGCCGTAGAGAAAT GCTGAGTCCCAGGGG TAAGAA.AAAGCAGTG


TGCCGTAGAGAAATG CTGAGTCCCAGGGGT AAGAAAAAGCAGTGT


GCCGTAGAGAAATGG TGAGTCCCAGGGGTG AGAAAAAGCAGTGTC


10CCGTAGAGAAATGGA GAGTCCCAGGGGTGT GAAAAAGCAGTGTCG


CGTAGAGAAATGGAA AGTCCCAGGGGTGTA AAAAAGCAGTGTCGC


GTAGAGAAATGGAAG GTCCCAGGGGTGTAC AAAAGCAGTGTCGCC


TAGAGAAATGGAAGA TCCCAGGGGTGTACA AAAGCAGTGTCGCCC


AGAGAAATGGAAGAC CCCAGGGGTGTACAC AAGCAGTGTCGCCCT


15GAGAAATGGAAGACA CCAGGGGTGTACACA AGCAGTGTCGCCCTT


AGAAATGGAAGACAC CAGGGGTGTACACAT GCAGTGTCGCCCTTC


GAAATGGAAGACACA AGGGGTGTACACATT CAGTGTCGCCCTTCC


AAATGGAAGACACAC GGGGTGTACACATTC AGTGTCGCCCTTCCA


AATGGAAGACACACT GGGTGTACACATTCC GTGTCGCCCTTCCAA


20ATGGAAGACACACTG GGTGTACACATTCCC TGTCGCCCTTCCAAA


TGGAAGACACACTGA GTGTACACATTCCCA GTCGCCCTTCCAAAG


GGAAGACACACTGAA TGTACACATTCCCAA TCGCCCTTCCAAAGG


GAAGACACACTGAAT GTACACATTCCCAAC CGCCCTTCCAAAGGC


AAGACACACTGAATC TACACATTCCCAACT GCCCTTCCAAAGGCA


25AGACACACTGAATCA ACACATTCCCAACTG CCCTTCCAAAGGCAG


GACACACTGAATCAC CACATTCCCAACTGT CCTTCCAAAGGCAGG


ACACACTGAATCACC ACATTCCCAACTGTG CTTCCAAAGGCAGGA


CACACTGAATCACCT CATTCCCAACTGTGA TTCCAAAGGCAGGAA


ACACTGAATCACCTG ATTCCCAACTGTGAC TCCAAAGGCAGGAAG


30CACTGAATCACCTGA TTCCCAACTGTGACA CCAAAGGCAGGAAGC


ACTGAATCACCTGAA TCCCAACTGTGACAA CAAAGGCAGGAAGCG


CTGAATCACCTGAAG CCCAACTGTGACAAG AAAGGCAGGAAGCGG


TGAATCACCTGAAGT CCAACTGTGACAAGA AAGGCAGGAAGCGGG


GAATCACCTGAAGTT CAACTGTGACAAGAA AGGCAGGAAGCGGGG


35AATCACCTGAAGTTC AACTGTGACAAGAAG GGCAGGAAGCGGGGC


ATCACCTGAAGTTCC ACTGTGACAAGAAGG GCAGGAAGCGGGGCT


TCACCTGAAGTTCCT CTGTGACAAGAAGGG CAGGAAGCGGGGCTT


CACCTGAAGTTCCTC TGTGACAAGAAGGGA AGGAAGCGGGGCTTC


ACCTGAAGTTCCTCA GTGACAAGAAGGGAT GGAAGCGGGGCTTCT


40CCTGAAGTTCCTCAA TGACAAGAAGGGATT GAAGCGGGGCTTCTG


CTGAAGTTCCTCAAT GACAAGAAGGGATTT AAGCGGGGCTTCTGC


TGAAGTTCCTCAATG ACAAGAAGGGATTTT AGCGGGGCTTCTGCT


GAAGTTCCTCAATGT CAAGAAGGGATTTTA GCGGGGCTTCTGCTG


AAGTTCCTCAATGTG AAGAAGGGATTTTAT CGGGGCTTCTGCTGG


45AGTTCCTCAATGTGC AGAAGGGATTTTATA GGGGCTTCTGCTGGT


GTTCCTCAATGTGCT GAAGGGATTTTATAA GGGCTTCTGCTGGTG


TTCCTCAATGTGCTG AAGGGATTTTATAAG GGCTTCTGCTGGTGT


TCCTCAATGTGCTGA AGGGATTTTATAAGA GCTTCTGCTGGTGTG


CCTCAATGTGCTGAG GGGATTTTATAAGAA CTTCTGCTGGTGTGT


50CTCAATGTGCTGAGT GGATTTTATAAGAAA TTCTGCTGGTGTGTG


TCAATGTGCTGAGTC GATTTTATAAGAAAA TCTGCTGGTGTGTGG




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-50-
CTGCTGGTGTGTGGA GGGGAAGGAGGACGT CGCAAGTTAATGTGG


TGCTGGTGTGTGGAT GGGAAGGAGGACGTG GCAAGTTAATGTGGA


GCTGGTGTGTGGATA GGAAGGAGGACGTGC CAAGTTAATGTGGAG


CTGGTGTGTGGATAA GAAGGAGGACGTGCA AAGTTAATGTGGAGC


S TGGTGTGTGGATAAG AAGGAGGACGTGCAC AGTTAATGTGGAGCT


GGTGTGTGGATAAGT AGGAGGACGTGCACT GTTAATGTGGAGCTC


GTGTGTGGATAAGTA GGAGGACGTGCACTG TTAATGTGGAGCTCA


TGTGTGGATAAGTAT GAGGACGTGCACTGC TAATGTGGAGCTCAA


GTGTGGATAAGTATG AGGACGTGCACTGCT AATGTGGAGCTCAAA


10TGTGGATAAGTATGG GGACGTGCACTGCTA ATGTGGAGCTCAAAT


GTGGATAAGTATGGG GACGTGCACTGCTAC TGTGGAGCTCAAATA


TGGATAAGTATGGGC ACGTGCACTGCTACA GTGGAGCTCAAATAT


GGATAAGTATGGGCA CGTGCACTGCTACAG TGGAGCTCAAATATG


GATAAGTATGGGCAG GTGCACTGCTACAGC GGAGCTCAAATATGC


15ATAAGTATGGGCAGC TGCACTGCTACAGCA GAGCTCAAATATGCC


TAAGTATGGGCAGCC GCACTGCTACAGCAT AGCTCAAATATGCCT


AAGTATGGGCAGCCT CACTGCTACAGCATG GCTCAAATATGCCTT


AGTATGGGCAGCCTC ACTGCTACAGCATGC CTCAAATATGCCTTA


GTATGGGCAGCCTCT CTGCTACAGCATGCA TCAAATATGCCTTAT


20TATGGGCAGCCTCTC TGCTACAGCATGCAG CAAATATGCCTTATT


ATGGGCAGCCTCTCC GCTACAGCATGCAGA AAATATGCCTTATTT


TGGGCAGCCTCTCCC CTACAGCATGCAGAG AATATGCCTTATTTT


GGGCAGCCTCTCCCA TACAGCATGCAGAGC ATATGCCTTATTTTG


GGCAGCCTCTCCCAG ACAGCATGCAGAGCA TATGCCTTATTTTGC


25GCAGCCTCTCCCAGG CAGCATGCAGAGCAA ATGCCTTATTTTGCA


CAGCCTCTCCCAGGC AGCATGCAGAGCAAG TGCCTTATTTTGCAC


AGCCTCTCCCAGGCT GCATGCAGAGCAAGT GCCTTATTTTGCACA


GCCTCTCCCAGGCTA CATGCAGAGCAAGTA CCTTATTTTGCACAA


CCTCTCCCAGGCTAC ATGCAGAGCAAGTAG CTTATTTTGCACAAA


30CTCTCCCAGGCTACA TGCAGAGCAAGTAGA TTATTTTGCACAAAA


TCTCCCAGGCTACAC GCAGAGCAAGTAGAC TATTTTGCACAAAAG


CTCCCAGGCTACACC CAGAGCAAGTAGACG ATTTTGCACAAAAGA


TCCCAGGCTACACCA AGAGCAAGTAGACGC TTTTGCACAAAAGAC


CCCAGGCTACACCAC GAGCAAGTAGACGCC TTTGCACAAAAGACT


35CCAGGCTACACCACC AGCAAGTAGACGCCT TTGCACAAAAGACTG


CAGGCTACACCACCA GCAAGTAGACGCCTG TGCACAAAAGACTGC


AGGCTACACCACCAA CAAGTAGACGCCTGC GCACAAAAGACTGCC


GGCTACACCACCAAG AAGTAGACGCCTGCC CACAAAAGACTGCCA


GCTACACCACCAAGG AGTAGACGCCTGCCG ACAAAAGACTGCCAA


40CTACACCACCAAGGG GTAGACGCCTGCCGC CAAAAGACTGCCAAG


TACACCACCAAGGGG TAGACGCCTGCCGCA AAAAGACTGCCAAGG


ACACCACCAAGGGGA AGACGCCTGCCGCAA AAAGACTGCCAAGGA


CACCACCAAGGGGAA GACGCCTGCCGCAAG AAGACTGCCAAGGAC


ACCACCAAGGGGAAG ACGCCTGCCGCAAGT AGACTGCCAAGGACA


45CCACCAAGGGGAAGG CGCCTGCCGCAAGTT GACTGCCAAGGACAT


CACCAAGGGGAAGGA GCCTGCCGCAAGTTA ACTGCCAAGGACATG


ACCAAGGGGAAGGAG CCTGCCGCAAGTTAA CTGCCAAGGACATGA


CCAAGGGGAAGGAGG CTGCCGCAAGTTAAT TGCCAAGGACATGAC


CAAGGGGAAGGAGGA TGCCGCAAGTTAATG GCCAAGGACATGACC


SOAAGGGGAAGGAGGAC GCCGCAAGTTAATGT CCAAGGACATGACCA


AGGGGAAGGAGGACG CCGCAAGTTAATGTG CAAGGACATGACCAG




CA 02376284 2001-12-05
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-51-
AAGGACATGACCAGC GTGAACTGATTTTTT TATGGTTTCTTTGAA


AGGACATGACCAGCA TGAACTGATTTTTTT ATGGTTTCTTTGAAT


GGACATGACCAGCAG GAACTGATTTTTTTT TGGTTTCTTTGAATG


GACATGACCAGCAGC AACTGATTTTTTTTA GGTTTCTTTGAATGG


ACATGACCAGCAGCT ACTGATTTTTTTTAA GTTTCTTTGAATGGT


CATGACCAGCAGCTG CTGATTTTTTTTAAA TTTCTTTGAATGGTA


ATGACCAGCAGCTGG TGATTTTTTTTAAAC TTCTTTGAATGGTAA


TGACCAGCAGCTGGC GATTTTTTTTAAACC TCTTTGAATGGTAAA


GACCAGCAGCTGGCT ATTTTTTTTAAACCA CTTTGAATGGTAAAC


10ACCAGCAGCTGGCTA TTTTTTTTAAACCAA TTTGAATGGTAAACT


CCAGCAGCTGGCTAC TTTTTTTAAACCAAA TTGAATGGTAAACTT


CAGCAGCTGGCTACA TTTTTTAAACCAAAG TGAATGGTAAACTTG


AGCAGCTGGCTACAG TTTTTAAACCAAAGT GAATGGTAAACTTGA


GCAGCTGGCTACAGC TTTTAAACCAAAGTT AATGGTAAACTTGAG


15CAGCTGGCTACAGCC TTTAAACCAAAGTTT ATGGTAAACTTGAGC


AGCTGGCTACAGCCT TTAAACCAAAGTTTA TGGTAAACTTGAGCA


GCTGGCTACAGCCTC TAAACCAAAGTTTAG GGTAAACTTGAGCAT


CTGGCTACAGCCTCG AAACCAAAGTTTAGA GTAAACTTGAGCATC


TGGCTACAGCCTCGA AACCAAAGTTTAGAA TAAACTTGAGCATCT


20GGCTACAGCCTCGAT ACCAAAGTTTAGAAA AAACTTGAGCATCTT


GCTACAGCCTCGATT CCAAAGTTTAGAAAG AACTTGAGCATCTTT


CTACAGCCTCGATTT CAAAGTTTAGAAAGA ACTTGAGCATCTTTT


TACAGCCTCGATTTA AAAGTTTAGAAAGAG CTTGAGCATCTTTTC


ACAGCCTCGATTTAT AAGTTTAGAAAGAGG TTGAGCATCTTTTCA


25CAGCCTCGATTTATA AGTTTAGAAAGAGGT TGAGCATCTTTTCAC


AGCCTCGATTTATAT GTTTAGAAAGAGGTT GAGCATCTTTTCACT


GCCTCGATTTATATT TTTAGAAAGAGGTTT AGCATCTTTTCACTT


CCTCGATTTATATTT TTAGAAAGAGGTTTT GCATCTTTTCACTTT


CTCGATTTATATTTC TAGAAAGAGGTTTTT CATCTTTTCACTTTC


30TCGATTTATATTTCT AGAAAGAGGTTTTTG ATCTTTTCACTTTCC


CGATTTATATTTCTG GAAAGAGGTTTTTGA TCTTTTCACTTTCCA


GATTTATATTTCTGT AAAGAGGTTTTTGAA CTTTTCACTTTCCAG


ATTTATATTTCTGTT AAGAGGTTTTTGAAA TTTTCACTTTCCAGT


TTTATATTTCTGTTT AGAGGTTTTTGAAAT TTTCACTTTCCAGTA


35TTATATTTCTGTTTG GAGGTTTTTGAAATG TTCACTTTCCAGTAG


TATATTTCTGTTTGT AGGTTTTTGAAATGC TCACTTTCCAGTAGT


ATATTTCTGTTTGTG GGTTTTTGAAATGCC CACTTTCCAGTAGTC


TATTTCTGTTTGTGG GTTTTTGAAATGCCT ACTTTCCAGTAGTCA


ATTTCTGTTTGTGGT TTTTTGAAATGCCTA CTTTCCAGTAGTCAG


40TTTCTGTTTGTGGTG TTTTGAAATGCCTAT TTTCCAGTAGTCAGC


TTCTGTTTGTGGTGA TTTGAAATGCCTATG TTCCAGTAGTCAGCA


TCTGTTTGTGGTGAA TTGAAATGCCTATGG TCCAGTAGTCAGCAA


CTGTTTGTGGTGAAC TGAAATGCCTATGGT CCAGTAGTCAGCAAA


TGTTTGTGGTGAACT GAAATGCCTATGGTT CAGTAGTCAGCAAAG


45GTTTGTGGTGAACTG AAATGCCTATGGTTT AGTAGTCAGCAAAGA


TTTGTGGTGAACTGA AATGCCTATGGTTTC GTAGTCAGCAAAGAG


TTGTGGTGAACTGAT ATGCCTATGGTTTCT TAGTCAGCAAAGAGC


TGTGGTGAACTGATT TGCCTATGGTTTCTT AGTCAGCAAAGAGCA


GTGGTGAACTGATTT GCCTATGGTTTCTTT GTCAGCAAAGAGCAG


50TGGTGAACTGATTTT CCTATGGTTTCTTTG TCAGCAAAGAGCAGT


GGTGAACTGATTTTT CTATGGTTTCTTTGA CAGCAAAGAGCAGTT




CA 02376284 2001-12-05
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-52-
AGCAAAGAGCAGTTT ACTCGAGCACAGCAC TTGGTCGAAGCGGCC


GCAAAGAGCAGTTTG CTCGAGCACAGCACC TGGTCGAAGCGGCCG


CAAAGAGCAGTTTGA TCGAGCACAGCACCC GGTCGAAGCGGCCGA


AAAGAGCAGTTTGAA CGAGCACAGCACCCA GTCGAAGCGGCCGAC


AAGAGCAGTTTGAAT GAGCACAGCACCCAG TCGAAGCGGCCGACC


AGAGCAGTTTGAATT AGCACAGCACCCAGA CGAAGCGGCCGACCA


GAGCAGTTTGAATTT GCACAGCACCCAGAC GAAGCGGCCGACCAC


AGCAGTTTGAATTTT CACAGCACCCAGACT AAGCGGCCGACCACT


GCAGTTTGAATTTTC ACAGCACCCAGACTT AGCGGCCGACCACTG


10CAGTTTGAATTTTCT CAGCACCCAGACTTC GCGGCCGACCACTGA


AGTTTGAATTTTCTT AGCACCCAGACTTCA CGGCCGACCACTGAC


GTTTGAATTTTCTTG GCACCCAGACTTCAT GGCCGACCACTGACT


TTTGAATTTTCTTGT CACCCAGACTTCATG GCCGACCACTGACTT


TTGAATTTTCTTGTC ACCCAGACTTCATGC CCGACCACTGACTTT


15TGAATTTTCTTGTCG CCCAGACTTCATGCG CGACCACTGACTTTG


GAATTTTCTTGTCGC CCAGACTTCATGCGC GACCACTGACTTTGT


AATTTTCTTGTCGCT CAGACTTCATGCGCC ACCACTGACTTTGTG


ATTTTCTTGTCGCTT AGACTTCATGCGCCC CCACTGACTTTGTGA


TTTTCTTGTCGCTTC GACTTCATGCGCCCG CACTGACTTTGTGAC


20TTTCTTGTCGCTTCC ACTTCATGCGCCCGT ACTGACTTTGTGACT


TTCTTGTCGCTTCCT CTTCATGCGCCCGTG CTGACTTTGTGACTT


TCTTGTCGCTTCCTA TTCATGCGCCCGTGG TGACTTTGTGACTTA


CTTGTCGCTTCCTAT TCATGCGCCCGTGGA GACTTTGTGACTTAG


TTGTCGCTTCCTATC CATGCGCCCGTGGAA ACTTTGTGACTTAGG


25TGTCGCTTCCTATCA ATGCGCCCGTGGAAT CTTTGTGACTTAGGC


GTCGCTTCCTATCAA TGCGCCCGTGGAATG TTTGTGACTTAGGCG


TCGCTTCCTATCAAA GCGCCCGTGGAATGC TTGTGACTTAGGCGG


CGCTTCCTATCAAAA CGCCCGTGGAATGCT TGTGACTTAGGCGGC


GCTTCCTATCAAAAT GCCCGTGGAATGCTC GTGACTTAGGCGGCT


30CTTCCTATCAAAATA CCCGTGGAATGCTCA TGACTTAGGCGGCTG


TTCCTATCAAAATAT CCGTGGAATGCTCAC GACTTAGGCGGCTGT


TCCTATCAAAATATT CGTGGAATGCTCACC ACTTAGGCGGCTGTG


CCTATCAAAATATTC GTGGAATGCTCACCA CTTAGGCGGCTGTGT


CTATCAAAATATTCA TGGAATGCTCACCAC TTAGGCGGCTGTGTT


35TATCAAAATATTCAG GGAATGCTCACCACA TAGGCGGCTGTGTTG


ATCAAAATATTCAGA GAATGCTCACCACAT AGGCGGCTGTGTTGC


TCAAAATATTCAGAG AATGCTCACCACATG GGCGGCTGTGTTGCC


CAAAATATTCAGAGA ATGCTCACCACATGT GCGGCTGTGTTGCCT


AAAATATTCAGAGAC TGCTCACCACATGTT CGGCTGTGTTGCCTA


40AAATATTCAGAGACT GCTCACCACATGTTG GGCTGTGTTGCCTAT


AATATTCAGAGACTC CTCACCACATGTTGG GCTGTGTTGCCTATG


ATATTCAGAGACTCG TCACCACATGTTGGT CTGTGTTGCCTATGT


TATTCAGAGACTCGA CACCACATGTTGGTC TGTGTTGCCTATGTA


ATTCAGAGACTCGAG ACCACATGTTGGTCG GTGTTGCCTATGTAG


45TTCAGAGACTCGAGC CCACATGTTGGTCGA TGTTGCCTATGTAGA


TCAGAGACTCGAGCA CACATGTTGGTCGAA GTTGCCTATGTAGAG


CAGAGACTCGAGCAC ACATGTTGGTCGAAG TTGCCTATGTAGAGA


AGAGACTCGAGCACA CATGTTGGTCGAAGC TGCCTATGTAGAGAA


GAGACTCGAGCACAG ATGTTGGTCGAAGCG GCCTATGTAGAGAAC


50AGACTCGAGCACAGC TGTTGGTCGAAGCGG CCTATGTAGAGAACA


GACTCGAGCACAGCA GTTGGTCGAAGCGGC CTATGTAGAGAACAC




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-53-
TATGTAGAGAACACG TATCGAGAATAGGAA ATGCTCCTGGAGCTC


ATGTAGAGAACACGC ATCGAGAATAGGAAA TGCTCCTGGAGCTCA


TGTAGAGAACACGCT TCGAGAATAGGAAAA GCTCCTGGAGCTCAC


GTAGAGAACACGCTT CGAGAATAGGAAAAC CTCCTGGAGCTCACA


TAGAGAACACGCTTC GAGAATAGGAAAACC TCCTGGAGCTCACAG


AGAGAACACGCTTCA AGAATAGGAAAACCT CCTGGAGCTCACAGC


GAGAACACGCTTCAC GAATAGGAAAACCTT CTGGAGCTCACAGCC


AGAACACGCTTCACC AATAGGAAAACCTTT TGGAGCTCACAGCCT


GAACACGCTTCACCC ATAGGAAAACCTTTA GGAGCTCACAGCCTT


10AACACGCTTCACCCC TAGGAAAACCTTTAA GAGCTCACAGCCTTC


ACACGCTTCACCCCC AGGAAAACCTTTAAA AGCTCACAGCCTTCT


CACGCTTCACCCCCA GGAAAACCTTTAAAC GCTCACAGCCTTCTG


ACGCTTCACCCCCAC GAAAACCTTTAAACC CTCACAGCCTTCTGT


CGCTTCACCCCCACT AAAACCTTTAAACCC TCACAGCCTTCTGTG


15GCTTCACCCCCACTC AAACCTTTAAACCCC CACAGCCTTCTGTGG


CTTCACCCCCACTCC AACCTTTAAACCCCG ACAGCCTTCTGTGGT


TTCACCCCCACTCCC ACCTTTAAACCCCGG CAGCCTTCTGTGGTG


TCACCCCCACTCCCC CCTTTAAACCCCGGT AGCCTTCTGTGGTGT


CACCCCCACTCCCCG CTTTAAACCCCGGTC GCCTTCTGTGGTGTC


20ACCCCCACTCCCCGT TTTAAACCCCGGTCA CCTTCTGTGGTGTCA


CCCCCACTCCCCGTA TTAAACCCCGGTCAT CTTCTGTGGTGTCAT


CCCCACTCCCCGTAC TAAACCCCGGTCATC TTCTGTGGTGTCATT


CCCACTCCCCGTACA AAACCCCGGTCATCC TCTGTGGTGTCATTT


CCACTCCCCGTACAG AACCCCGGTCATCCG CTGTGGTGTCATTTC


25CACTCCCCGTACAGT ACCCCGGTCATCCGG TGTGGTGTCATTTCT


ACTCCCCGTACAGTG CCCCGGTCATCCGGA GTGGTGTCATTTCTG


CTCCCCGTACAGTGC CCCGGTCATCCGGAC TGGTGTCATTTCTGA


TCCCCGTACAGTGCG CCGGTCATCCGGACA GGTGTCATTTCTGAA


CCCCGTACAGTGCGC CGGTCATCCGGACAT GTGTCATTTCTGAAA


30CCCGTACAGTGCGCA GGTCATCCGGACATC TGTCATTTCTGAAAC


CCGTACAGTGCGCAC GTCATCCGGACATCC GTCATTTCTGAAACA


CGTACAGTGCGCACA TCATCCGGACATCCC TCATTTCTGAAACAA


GTACAGTGCGCACAG CATCCGGACATCCCA CATTTCTGAAACAAG


TACAGTGCGCACAGG ATCCGGACATCCCAA ATTTCTGAAACAAGG


35ACAGTGCGCACAGGC TCCGGACATCCCAAC TTTCTGAAACAAGGG


CAGTGCGCACAGGCT CCGGACATCCCAACG TTCTGAAACAAGGGC


AGTGCGCACAGGCTT CGGACATCCCAACGC TCTGAAACAAGGGCG


GTGCGCACAGGCTTT GGACATCCCAACGCA CTGAAACAAGGGCGT


TGCGCACAGGCTTTA GACATCCCAACGCAT TGAAACAAGGGCGTG


40GCGCACAGGCTTTAT ACATCCCAACGCATG GAAACAAGGGCGTGG


CGCACAGGCTTTATC CATCCCAACGCATGC AAACAAGGGCGTGGA


GCACAGGCTTTATCG ATCCCAACGCATGCT AACAAGGGCGTGGAT


CACAGGCTTTATCGA TCCCAACGCATGCTC ACAAGGGCGTGGATC


ACAGGCTTTATCGAG CCCAACGCATGCTCC CAAGGGCGTGGATCC


45CAGGCTTTATCGAGA CCAACGCATGCTCCT AAGGGCGTGGATCCC


AGGCTTTATCGAGAA CAACGCATGCTCCTG AGGGCGTGGATCCCT


GGCTTTATCGAGAAT AACGCATGCTCCTGG GGGCGTGGATCCCTC


GCTTTATCGAGAATA ACGCATGCTCCTGGA GGCGTGGATCCCTCA


CTTTATCGAGAATAG CGCATGCTCCTGGAG GCGTGGATCCCTCAA


50TTTATCGAGAATAGG GCATGCTCCTGGAGC CGTGGATCCCTCAAC


TTATCGAGAATAGGA CATGCTCCTGGAGCT GTGGATCCCTCAACC




CA 02376284 2001-12-05
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-S4-
TGGATCCCTCAACCA TTGGGGACTATTGGA GTATCTAAGAATGTT


GGATCCCTCAACCAA TGGGGACTATTGGAG TATCTAAGAATGTTC


GATCCCTCAACCAAG GGGGACTATTGGAGA ATCTAAGAATGTTCT


ATCCCTCAACCAAGA GGGACTATTGGAGAA TCTAAGAATGTTCTA


S TCCCTCAACCAAGAA GGACTATTGGAGAAA CTAAGAATGTTCTAG


CCCTCAACCAAGAAG GACTATTGGAGAAAA TAAGAATGTTCTAGG


CCTCAACCAAGAAGA ACTATTGGAGAAAAT AAGAATGTTCTAGGG


CTCAACCAAGAAGAA CTATTGGAGAAAATA AGAATGTTCTAGGGC


TCAACCAAGAAGAAT TATTGGAGAAAATAA GAATGTTCTAGGGCA


10CAACCAAGAAGAATG ATTGGAGAAAATAAG AATGTTCTAGGGCAC


AACCAAGAAGAATGT TTGGAGAAAATAAGG ATGTTCTAGGGCACT


ACCAAGAAGAATGTT TGGAGAAAATAAGGT TGTTCTAGGGCACTC


CCAAGAAGAATGTTT GGAGAAAATAAGGTG GTTCTAGGGCACTCT


CAAGAAGAATGTTTA GAGAAAATAAGGTGG TTCTAGGGCACTCTG


1SAAGAAGAATGTTTAT AGAAAATAAGGTGGA TCTAGGGCACTCTGG


AGAAGAATGTTTATG GAAAATAAGGTGGAG CTAGGGCACTCTGGG


GAAGAATGTTTATGT AAAATAAGGTGGAGT TAGGGCACTCTGGGA


AAGAATGTTTATGTC AAATAAGGTGGAGTC AGGGCACTCTGGGAA


AGAATGTTTATGTCT AATAAGGTGGAGTCC GGGCACTCTGGGAAC


20GAATGTTTATGTCTT ATAAGGTGGAGTCCT GGCACTCTGGGAACC


AATGTTTATGTCTTC TAAGGTGGAGTCCTA GCACTCTGGGAACCT


ATGTTTATGTCTTCA AAGGTGGAGTCCTAC CACTCTGGGAACCTA


TGTTTATGTCTTCAA AGGTGGAGTCCTACT ACTCTGGGAACCTAT


GTTTATGTCTTCAAG GGTGGAGTCCTACTT CTCTGGGAACCTATA


25TTTATGTCTTCAAGT GTGGAGTCCTACTTG TCTGGGAACCTATAA


TTATGTCTTCAAGTG TGGAGTCCTACTTGT CTGGGAACCTATAAA


TATGTCTTCAAGTGA GGAGTCCTACTTGTT TGGGAACCTATAAAG


ATGTCTTCAAGTGAC GAGTCCTACTTGTTT GGGAACCTATAAAGG


TGTCTTCAAGTGACC AGTCCTACTTGTTTA GGAACCTATAAAGGC


30GTCTTCAAGTGACCT GTCCTACTTGTTTAA GAACCTATAAAGGCA


TCTTCAAGTGACCTG TCCTACTTGTTTAAA AACCTATAAAGGCAG


CTTCAAGTGACCTGT CCTACTTGTTTAAAA ACCTATAAAGGCAGG


TTCAAGTGACCTGTA CTACTTGTTTAAAAA CCTATAAAGGCAGGT


TCAAGTGACCTGTAC TACTTGTTTAAAAAA CTATAAAGGCAGGTA


3SCAAGTGACCTGTACT ACTTGTTTAAAAAAT TATAAAGGCAGGTAT


AAGTGACCTGTACTG CTTGTTTAAAAAATA ATAAAGGCAGGTATT


AGTGACCTGTACTGC TTGTTTAAAAAATAT TAAAGGCAGGTATTT


GTGACCTGTACTGCT TGTTTAAAAAATATG AAAGGCAGGTATTTC


TGACCTGTACTGCTT GTTTAAAAAATATGT AAGGCAGGTATTTCG


40GACCTGTACTGCTTG TTTAA.AA.A.ATATGTA AGGCAGGTATTTCGG


ACCTGTACTGCTTGG TTAAAAAATATGTAT GGCAGGTATTTCGGG


CCTGTACTGCTTGGG TAAAAAATATGTATC GCAGGTATTTCGGGC


CTGTACTGCTTGGGG AAAAAATATGTATCT CAGGTATTTCGGGCC


TGTACTGCTTGGGGA AAAAATATGTATCTA AGGTATTTCGGGCCC


4SGTACTGCTTGGGGAC AAAATATGTATCTAA GGTATTTCGGGCCCT


TACTGCTTGGGGACT AAATATGTATCTAAG GTATTTCGGGCCCTC


ACTGCTTGGGGACTA AATATGTATCTAAGA TATTTCGGGCCCTCC


CTGCTTGGGGACTAT ATATGT'ATCTAAGAA ATTTCGGGCCCTCCT


TGCTTGGGGACTATT TATGTATCTAAGAAT TTTCGGGCCCTCCTC


SOGCTTGGGGACTATTG ATGTATCTAAGAATG TTCGGGCCCTCCTCT


CTTGGGGACTATTGG TGTATCTAAGAATGT TCGGGCCCTCCTCTT




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-55-
CGGGCCCTCCTCTTC CAGGATGGCTTTTGC AGAGTCAGCCTCCAC


GGGCCCTCCTCTTCA AGGATGGCTTTTGCT GAGTCAGCCTCCACA


GGCCCTCCTCTTCAG GGATGGCTTTTGCTG AGTCAGCCTCCACAT


GCCCTCCTCTTCAGG GATGGCTTTTGCTGC GTCAGCCTCCACATT


CCCTCCTCTTCAGGA ATGGCTTTTGCTGCG TCAGCCTCCACATTC


CCTCCTCTTCAGGAA TGGCTTTTGCTGCGG CAGCCTCCACATTCA


CTCCTCTTCAGGAAT GGCTTTTGCTGCGGC AGCCTCCACATTCAG


TCCTCTTCAGGAATC GCTTTTGCTGCGGCC GCCTCCACATTCAGA


CCTCTTCAGGAATCT CTTTTGCTGCGGCCC CCTCCACATTCAGAG


10CTCTTCAGGAATCTT TTTTGCTGCGGCCCC CTCCACATTCAGAGG


TCTTCAGGAATCTTC TTTGCTGCGGCCCCG TCCACATTCAGAGGC


CTTCAGGAATCTTCC TTGCTGCGGCCCCGT CCACATTCAGAGGCA


TTCAGGAATCTTCCT TGCTGCGGCCCCGTG CACATTCAGAGGCAT


TCAGGAATCTTCCTG GCTGCGGCCCCGTGG ACATTCAGAGGCATC


15CAGGAATCTTCCTGA CTGCGGCCCCGTGGG CATTCAGAGGCATCA


AGGAATCTTCCTGAA TGCGGCCCCGTGGGG ATTCAGAGGCATCAC


GGAATCTTCCTGAAG GCGGCCCCGTGGGGT TTCAGAGGCATCACA


GAATCTTCCTGAAGA CGGCCCCGTGGGGTA TCAGAGGCATCACAA


AATCTTCCTGAAGAC GGCCCCGTGGGGTAG CAGAGGCATCACAAG


20ATCTTCCTGAAGACA GCCCCGTGGGGTAGG AGAGGCATCACAAGT


TCTTCCTGAAGACAT CCCCGTGGGGTAGGA GAGGCATCACAAGTA


CTTCCTGAAGACATG CCCGTGGGGTAGGAG AGGCATCACAAGTAA


TTCCTGAAGACATGG CCGTGGGGTAGGAGG GGCATCACAAGTAAT


TCCTGAAGACATGGC CGTGGGGTAGGAGGG GCATCACAAGTAATG


25CCTGAAGACATGGCC GTGGGGTAGGAGGGA CATCACAAGTAATGG


CTGAAGACATGGCCC TGGGGTAGGAGGGAC ATCACAAGTAATGGC


TGAAGACATGGCCCA GGGGTAGGAGGGACA TCACAAGTAATGGCA


GAAGACATGGCCCAG GGGTAGGAGGGACAG CACAAGTAATGGCAC


AAGACATGGCCCAGT GGTAGGAGGGACAGA ACAAGTAATGGCACA


30AGACATGGCCCAGTC GTAGGAGGGACAGAG CAAGTAATGGCACAA


GACATGGCCCAGTCG TAGGAGGGACAGAGA AAGTAATGGCACAAT


ACATGGCCCAGTCGA AGGAGGGACAGAGAG AGTAATGGCACAATT


CATGGCCCAGTCGAA GGAGGGACAGAGAGA GTAATGGCACAATTC


ATGGCCCAGTCGAAG GAGGGACAGAGAGAC TAATGGCACAATTCT


35TGGCCCAGTCGAAGG AGGGACAGAGAGACG AATGGCACAATTCTT


GGCCCAGTCGAAGGC GGGACAGAGAGACGG ATGGCACAATTCTTC


GCCCAGTCGAAGGCC GGACAGAGAGACGGG TGGCACAATTCTTCG


CCCAGTCGAAGGCCC GACAGAGAGACGGGA GGCACAATTCTTCGG


CCAGTCGAAGGCCCA ACAGAGAGACGGGAG GCACAATTCTTCGGA


40CAGTCGAAGGCCCAG CAGAGAGACGGGAGA CACAATTCTTCGGAT


AGTCGAAGGCCCAGG AGAGAGACGGGAGAG ACAATTCTTCGGATG


GTCGAAGGCCCAGGA GAGAGACGGGAGAGT CAATTCTTCGGATGA


TCGAAGGCCCAGGAT AGAGACGGGAGAGTC AATTCTTCGGATGAC


CGAAGGCCCAGGATG GAGACGGGAGAGTCA ATTCTTCGGATGACT


45GAAGGCCCAGGATGG AGACGGGAGAGTCAG TTCTTCGGATGACTG


AAGGCCCAGGATGGC GACGGGAGAGTCAGC TCTTCGGATGACTGC


AGGCCCAGGATGGCT ACGGGAGAGTCAGCC CTTCGGATGACTGCA


GGCCCAGGATGGCTT CGGGAGAGTCAGCCT TTCGGATGACTGCAG


GCCCAGGATGGCTTT GGGAGAGTCAGCCTC TCGGATGACTGCAGA


50CCCAGGATGGCTTTT GGAGAGTCAGCCTCC CGGATGACTGCAGAA


CCAGGATGGCTTTTG GAGAGTCAGCCTCCA GGATGACTGCAGAAA




CA 02376284 2001-12-05
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-56-
GATGACTGCAGAAAA ATTTCTGAGGATAAG TTTTGTCCTCCTTAG


ATGACTGCAGAAAAT TTTCTGAGGATAAGC TTTGTCCTCCTTAGC


TGACTGCAGAAAATA TTCTGAGGATAAGCT TTGTCCTCCTTAGCA


GACTGCAGAAAATAG TCTGAGGATAAGCTC TGTCCTCCTTAGCAC


ACTGCAGAAAATAGT CTGAGGATAAGCTCT GTCCTCCTTAGCACA


CTGCAGAAAATAGTG TGAGGATAAGCTCTT TCCTCCTTAGCACAA


TGCAGAAAATAGTGT GAGGATAAGCTCTTT CCTCCTTAGCACAAT


GCAGAAAATAGTGTT AGGATAAGCTCTTTA CTCCTTAGCACAATG


CAGAAAATAGTGTTT GGATAAGCTCTTTAA TCCTTAGCACAATGT


10AGAAAATAGTGTTTT GATAAGCTCTTTAAA CCTTAGCACAATGTA


GAAAATAGTGTTTTG ATAAGCTCTTTAAAG CTTAGCACAATGTAA


AAAATAGTGTTTTGT TAAGCTCTTTAAAGG TTAGCACAATGTAAA


AAATAGTGTTTTGTA AAGCTCTTTAAAGGC TAGCACAATGTAAAA


AATAGTGTTTTGTAG AGCTCTTTAAAGGCA AGCACAATGTAAAAA


15ATAGTGTTTTGTAGT GCTCTTTAAAGGCAA GCACAATGTAAAAAA


TAGTGTTTTGTAGTT CTCTTTAAAGGCAAA CACAATGTAAAAAAG


AGTGTTTTGTAGTTC TCTTTAAAGGCAAAG ACAATGTAAAAAAGA


GTGTTTTGTAGTTCA CTTTAAAGGCAAAGC CAATGTAAAAAAGAA


TGTTTTGTAGTTCAA TTTAAAGGCAAAGCT AATGTAAAAAAGAAT


20GTTTTGTAGTTCAAC TTAAAGGCAAAGCTT ATGTAAAAAAGAATA


TTTTGTAGTTCAACA TAAAGGCAAAGCTTT TGTAAAAAAGAATAG


TTTGTAGTTCAACAA AAAGGCAAAGCTTTA GTAAAA.AAGAATAGT


TTGTAGTTCAACAAC AAGGCAAAGCTTTAT TAAAAAAGAATAGTA


TGTAGTTCAACAACT AGGCAAAGCTTTATT AAAAAAGAATAGTAA


25GTAGTTCAACAACTC GGCAAAGCTTTATTT AAAAAGAATAGTAAT


TAGTTCAACAACTCA GCAAAGCTTTATTTT AAAAGAATAGTAATA


AGTTCAACAACTCAA CAAAGCTTTATTTTC AAAGAATAGTAATAT


GTTCAACAACTCAAG AAAGCTTTATTTTCA AAGAATAGTAATATC


TTCAACAACTCAAGA AAGCTTTATTTTCAT AGAATAGTAATATCA


30TCAACAACTCAAGAC AGCTTTATTTTCATC GAATAGTAATATCAG


CAACAACTCAAGACG GCTTTATTTTCATCT AATAGTAATATCAGA


AACAACTCAAGACGA CTTTATTTTCATCTC ATAGTAATATCAGAA


ACAACTCAAGACGAA TTTATTTTCATCTCT TAGTAATATCAGAAC


CAACTCAAGACGAAG TTATTTTCATCTCTC AGTAATATCAGAACA


35AACTCAAGACGAAGC TATTTTCATCTCTCA GTAATATCAGAACAG


ACTCAAGACGAAGCT ATTTTCATCTCTCAT TAATATCAGAACAGG


CTCAAGACGAAGCTT TTTTCATCTCTCATC AATATCAGAACAGGA


TCAAGACGAAGCTTA TTTCATCTCTCATCT ATATCAGAACAGGAA


CAAGACGAAGCTTAT TTCATCTCTCATCTT TATCAGAACAGGAAG


40AAGACGAAGCTTATT TCATCTCTCATCTTT ATCAGAACAGGAAGG


AGACGAAGCTTATTT CATCTCTCATCTTTT TCAGAACAGGAAGGA


GACGAAGCTTATTTC ATCTCTCATCTTTTG CAGAACAGGAAGGAG


ACGAAGCTTATTTCT TCTCTCATCTTTTGT AGAACAGGAAGGAGG


CGAAGCTTATTTCTG CTCTCATCTTTTGTC GAACAGGAAGGAGGA


45GAAGCTTATTTCTGA TCTCATCTTTTGTCC AACAGGAAGGAGGAA


AAGCTTATTTCTGAG CTCATCTTTTGTCCT ACAGGAAGGAGGAAT


AGCTTATTTCTGAGG TCATCTTTTGTCCTC CAGGAAGGAGGAATG


GCTTATTTCTGAGGA CATCTTTTGTCCTCC AGGAAGGAGGAATGG


CTTATTTCTGAGGAT ATCTTTTGTCCTCCT GGAAGGAGGAATGGC


50TTATTTCTGAGGATA TCTTTTGTCCTCCTT GAAGGAGGAATGGCT


TATTTCTGAGGATAA CTTTTGTCCTCCTTA AAGGAGGAATGGCTT




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-57-
AGGAGGAATGGCTTG GATTCACCCATGTTT ATTCACACATATATG


GGAGGAATGGCTTGC ATTCACCCATGTTTG TTCACACATATATGC


GAGGAATGGCTTGCT TTCACCCATGTTTGT TCACACATATATGCA


AGGAATGGCTTGCTG TCACCCATGTTTGTT CACACATATATGCAG


GGAATGGCTTGCTGG CACCCATGTTTGTTG ACACATATATGCAGA


GAATGGCTTGCTGGG ACCCATGTTTGTTGA CACATATATGCAGAG


AATGGCTTGCTGGGG CCCATGTTTGTTGAA ACATATATGCAGAGA


ATGGCTTGCTGGGGA CCATGTTTGTTGAAC CATATATGCAGAGAA


TGGCTTGCTGGGGAG CATGTTTGTTGAACT ATATATGCAGAGAAG


10GGCTTGCTGGGGAGC ATGTTTGTTGAACTT TATATGCAGAGAAGA


GCTTGCTGGGGAGCC TGTTTGTTGAACTTA ATATGCAGAGAAGAT


CTTGCTGGGGAGCCC GTTTGTTGAACTTAG TATGCAGAGAAGATA


TTGCTGGGGAGCCCA TTTGTTGAACTTAGA ATGCAGAGAAGATAT


TGCTGGGGAGCCCAT TTGTTGAACTTAGAG TGCAGAGAAGATATG


15GCTGGGGAGCCCATC TGTTGAACTTAGAGT GCAGAGAAGATATGT


CTGGGGAGCCCATCC GTTGAACTTAGAGTC CAGAGAAGATATGTT


TGGGGAGCCCATCCA TTGAACTTAGAGTCA AGAGAAGATATGTTC


GGGGAGCCCATCCAG TGAACTTAGAGTCAT GAGAAGATATGTTCT


GGGAGCCCATCCAGG GAACTTAGAGTCATT AGAAGATATGTTCTT


20GGAGCCCATCCAGGA AACTTAGAGTCATTC GAAGATATGTTCTTG


GAGCCCATCCAGGAC ACTTAGAGTCATTCT AAGATATGTTCTTGT


AGCCCATCCAGGACA CTTAGAGTCATTCTC AGATATGTTCTTGTT


GCCCATCCAGGACAC TTAGAGTCATTCTCA GATATGTTCTTGTTA


CCCATCCAGGACACT TAGAGTCATTCTCAT ATATGTTCTTGTTAA


25CCATCCAGGACACTG AGAGTCATTCTCATG TATGTTCTTGTTAAC


CATCCAGGACACTGG GAGTCATTCTCATGC ATGTTCTTGTTAACA


ATCCAGGACACTGGG AGTCATTCTCATGCT TGTTCTTGTTAACAT


TCCAGGACACTGGGA GTCATTCTCATGCTT GTTCTTGTTAACATT


CCAGGACACTGGGAG TCATTCTCATGCTTT TTCTTGTTAACATTG


30CAGGACACTGGGAGC CATTCTCATGCTTTT TCTTGTTAACATTGT


AGGACACTGGGAGCA ATTCTCATGCTTTTC CTTGTTAACATTGTA


GGACACTGGGAGCAC TTCTCATGCTTTTCT TTGTTAACATTGTAT


GACACTGGGAGCACA TCTCATGCTTTTCTT TGTTAACATTGTATA


ACACTGGGAGCACAT CTCATGCTTTTCTTT GTTAACATTGTATAC


35CACTGGGAGCACATA TCATGCTTTTCTTTA TTAACATTGTATACA


ACTGGGAGCACATAG CATGCTTTTCTTTAT TAACATTGTATACAA


CTGGGAGCACATAGA ATGCTTTTCTTTATA AACATTGTATACAAC


TGGGAGCACATAGAG TGCTTTTCTTTATAA ACATTGTATACAACA


GGGAGCACATAGAGA GCTTTTCTTTATAAT CATTGTATACAACAT


40GGAGCACATAGAGAT CTTTTCTTTATAATT ATTGTATACAACATA


GAGCACATAGAGATT TTTTCTTTATAATTC TTGTATACAACATAG


AGCACATAGAGATTC TTTCTTTATAATTCA TGTATACAACATAGC


GCACATAGAGATTCA TTCTTTATAATTCAC GTATACAACATAGCC


CACATAGAGATTCAC TCTTTATAATTCACA TATACAACATAGCCC


45ACATAGAGATTCACC CTTTATAATTCACAC ATACAACATAGCCCC


CATAGAGATTCACCC TTTATAATTCACACA TACAACATAGCCCCA


ATAGAGATTCACCCA TTATAATTCACACAT ACAACATAGCCCCAA


TAGAGATTCACCCAT TATAATTCACACATA CAACATAGCCCCAAA


AGAGATTCACCCATG ATAATTCACACATAT AACATAGCCCCAAAT


50GAGATTCACCCATGT TAATTCACACATATA ACATAGCCCCAAATA


AGATTCACCCATGTT AATTCACACATATAT CATAGCCCCAAATAT




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-58-
ATAGCCCCAAATATA AGAGATGCTATATGA CCCAGAGACTGGGCT


TAGCCCCAAATATAG GAGATGCTATATGAT CCAGAGACTGGGCTG


AGCCCCAAATATAGT AGATGCTATATGATA CAGAGACTGGGCTGC


GCCCCAAATATAGTA GATGCTATATGATAC AGAGACTGGGCTGCT


CCCCAAATATAGTAA ATGCTATATGATACA GAGACTGGGCTGCTC


CCCAAATATAGTAAG TGCTATATGATACAA AGACTGGGCTGCTCT


CCAAATATAGTAAGA GCTATATGATACAAC GACTGGGCTGCTCTC


CAAATATAGTAAGAT CTATATGATACAACT ACTGGGCTGCTCTCC


AAATATAGTAAGATC TATATGATACAACTG CTGGGCTGCTCTCCC


10AATATAGTAAGATCT ATATGATACAACTGT TGGGCTGCTCTCCCG


ATATAGTAAGATCTA TATGATACAACTGTG GGGCTGCTCTCCCGG


TATAGTAAGATCTAT ATGATACAACTGTGG GGCTGCTCTCCCGGA


ATAGTAAGATCTATA TGATACAACTGTGGC GCTGCTCTCCCGGAG


TAGTAAGATCTATAC GATACAACTGTGGCC CTGCTCTCCCGGAGG


15AGTAAGATCTATACT ATACAACTGTGGCCA TGCTCTCCCGGAGGC


GTAAGATCTATACTA TACAACTGTGGCCAT GCTCTCCCGGAGGCC


TAAGATCTATACTAG ACAACTGTGGCCATG CTCTCCCGGAGGCCA


AAGATCTATACTAGA CAACTGTGGCCATGA TCTCCCGGAGGCCAA


AGATCTATACTAGAT AACTGTGGCCATGAC CTCCCGGAGGCCAAA


20GATCTATACTAGATA ACTGTGGCCATGACT TCCCGGAGGCCAAAC


ATCTATACTAGATAA CTGTGGCCATGACTG CCCGGAGGCCAAACC


TCTATACTAGATAAT TGTGGCCATGACTGA CCGGAGGCCAAACCC


CTATACTAGATAATC GTGGCCATGACTGAG CGGAGGCCAAACCCA


TATACTAGATAATCC TGGCCATGACTGAGG GGAGGCCAAACCCAA


25ATACTAGATAATCCT GGCCATGACTGAGGA GAGGCCAAACCCAAG


TACTAGATAATCCTA GCCATGACTGAGGAA AGGCCAAACCCAAGA


ACTAGATAATCCTAG CCATGACTGAGGAAA GGCCAAACCCAAGAA


CTAGATAATCCTAGA CATGACTGAGGAAAG GCCAAACCCAAGAAG


TAGATAATCCTAGAT ATGACTGAGGAAAGG CCAAACCCAAGAAGG


30AGATAATCCTAGATG TGACTGAGGAAAGGA CAAACCCAAGAAGGT


GATAATCCTAGATGA GACTGAGGAAAGGAG AAACCCAAGAAGGTC


ATAATCCTAGATGAA ACTGAGGAAAGGAGC AACCCAAGAAGGTCT


TAATCCTAGATGAAA CTGAGGAAAGGAGCT ACCCAAGAAGGTCTG


AATCCTAGATGAAAT TGAGGAAAGGAGCTC CCCAAGAAGGTCTGG


35ATCCTAGATGAAATG GAGGAAAGGAGCTCA CCAAGAAGGTCTGGC


TCCTAGATGAAATGT AGGAAAGGAGCTCAC CAAGAAGGTCTGGCA


CCTAGATGAAATGTT GGAAAGGAGCTCACG AAGAAGGTCTGGCAA


CTAGATGAAATGTTA GAAAGGAGCTCACGC AGAAGGTCTGGCAAA


TAGATGAAATGTTAG AAAGGAGCTCACGCC GAAGGTCTGGCAAAG


40AGATGAAATGTTAGA AAGGAGCTCACGCCC AAGGTCTGGCAAAGT


GATGAAATGTTAGAG AGGAGCTCACGCCCA AGGTCTGGCAAAGTC


ATGAAATGTTAGAGA GGAGCTCACGCCCAG GGTCTGGCAAAGTCA


TGAAATGTTAGAGAT GAGCTCACGCCCAGA GTCTGGCAAAGTCAG


GAAATGTTAGAGATG AGCTCACGCCCAGAG TCTGGCAAAGTCAGG


45AAATGTTAGAGATGC GCTCACGCCCAGAGA CTGGCAAAGTCAGGC


AATGTTAGAGATGCT CTCACGCCCAGAGAC TGGCAAAGTCAGGCT


ATGTTAGAGATGCTA TCACGCCCAGAGACT GGCAAAGTCAGGCTC


TGTTAGAGATGCTAT CACGCCCAGAGACTG GCAAAGTCAGGCTCA


GTTAGAGATGCTATA ACGCCCAGAGACTGG CAAAGTCAGGCTCAG


50TTAGAGATGCTATAT CGCCCAGAGACTGGG AAAGTCAGGCTCAGG


TAGAGATGCTATATG GCCCAGAGACTGGGC AAGTCAGGCTCAGGG




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-59-
AGTCAGGCTCAGGGA GCTGCATAGAGCTCT CCTATTAGCTTTTCT


GTCAGGCTCAGGGAG CTGCATAGAGCTCTC CTATTAGCTTTTCTT


TCAGGCTCAGGGAGA TGCATAGAGCTCTCC TATTAGCTTTTCTTT


CAGGCTCAGGGAGAC GCATAGAGCTCTCCT ATTAGCTTTTCTTTA


AGGCTCAGGGAGACT CATAGAGCTCTCCTT TTAGCTTTTCTTTAT


GGCTCAGGGAGACTC ATAGAGCTCTCCTTG TAGCTTTTCTTTATT


GCTCAGGGAGACTCT TAGAGCTCTCCTTGA AGCTTTTCTTTATTT


CTCAGGGAGACTCTG AGAGCTCTCCTTGAA GCTTTTCTTTATTTT


TCAGGGAGACTCTGC GAGCTCTCCTTGAAA CTTTTCTTTATTTTT


10CAGGGAGACTCTGCC AGCTCTCCTTGAAAA TTTTCTTTATTTTTT


AGGGAGACTCTGCCC GCTCTCCTTGAAAAC TTTCTTTATTTTTTT


GGGAGACTCTGCCCT CTCTCCTTGAAAACA TTCTTTATTTTTTTA


GGAGACTCTGCCCTG TCTCCTTGAAAACAG TCTTTATTTTTTTAA


GAGACTCTGCCCTGC CTCCTTGAAAACAGA CTTTATTTTTTTAAC


15AGACTCTGCCCTGCT TCCTTGAAAACAGAG TTTATTTTTTTAACT


GACTCTGCCCTGCTG CCTTGAAAACAGAGG TTATTTTTTTAACTT


ACTCTGCCCTGCTGC CTTGAAAACAGAGGG TATTTTTTTAACTTT


CTCTGCCCTGCTGCA TTGAAAACAGAGGGG ATTTTTTTAACTTTT


TCTGCCCTGCTGCAG TGAAAACAGAGGGGT TTTTTTTAACTTTTT


20CTGCCCTGCTGCAGA GAAAACAGAGGGGTC TTTTTTAACTTTTTG


TGCCCTGCTGCAGAC AAAACAGAGGGGTCT TTTTTAACTTTTTGG


GCCCTGCTGCAGACC AAACAGAGGGGTCTC TTTTAACTTTTTGGG


CCCTGCTGCAGACCT AACAGAGGGGTCTCA TTTAACTTTTTGGGG


CCTGCTGCAGACCTC ACAGAGGGGTCTCAA TTAACTTTTTGGGGG


25CTGCTGCAGACCTCG CAGAGGGGTCTCAAG TAACTTTTTGGGGGG


TGCTGCAGACCTCGG AGAGGGGTCTCAAGA AACTTTTTGGGGGGA


GCTGCAGACCTCGGT GAGGGGTCTCAAGAC ACTTTTTGGGGGGAA


CTGCAGACCTCGGTG AGGGGTCTCAAGACA CTTTTTGGGGGGAAA


TGCAGACCTCGGTGT GGGGTCTCAAGACAT TTTTTGGGGGGAAAA


30GCAGACCTCGGTGTG GGGTCTCAAGACATT TTTTGGGGGGAAAAG


CAGACCTCGGTGTGG GGTCTCAAGACATTC TTTGGGGGGAAAAGT


AGACCTCGGTGTGGA GTCTCAAGACATTCT TTGGGGGGAAAAGTA


GACCTCGGTGTGGAC TCTCAAGACATTCTG TGGGGGGAAAAGTAT


ACCTCGGTGTGGACA CTCAAGACATTCTGC GGGGGGAAAAGTATT


35CCTCGGTGTGGACAC TCAAGACATTCTGCC GGGGGAAAAGTATTT


CTCGGTGTGGACACA CAAGACATTCTGCCT GGGGAAAAGTATTTT


TCGGTGTGGACACAC AAGACATTCTGCCTA GGGAAAAGTATTTTT


CGGTGTGGACACACG AGACATTCTGCCTAC GGAAAAGTATTTTTG


GGTGTGGACACACGC GACATTCTGCCTACC GAAAAGTATTTTTGA


40GTGTGGACACACGCT ACATTCTGCCTACCT AAAAGTATTTTTGAG


TGTGGACACACGCTG CATTCTGCCTACCTA AAAGTATTTTTGAGA


GTGGACACACGCTGC ATTCTGCCTACCTAT AAGTATTTTTGAGAA


TGGACACACGCTGCA TTCTGCCTACCTATT AGTATTTTTGAGAAG


GGACACACGCTGCAT TCTGCCTACCTATTA GTATTTTTGAGAAGT


45GACACACGCTGCATA CTGCCTACCTATTAG TATTTTTGAGAAGTT


ACACACGCTGCATAG TGCCTACCTATTAGC ATTTTTGAGAAGTTT


CACACGCTGCATAGA GCCTACCTATTAGCT TTTTTGAGAAGTTTG


ACACGCTGCATAGAG CCTACCTATTAGCTT TTTTGAGAAGTTTGT


CACGCTGCATAGAGC CTACCTATTAGCTTT TTTGAGAAGTTTGTC


50ACGCTGCATAGAGCT TACCTATTAGCTTTT TTGAGAAGTTTGTCT


CGCTGCATAGAGCTC ACCTATTAGCTTTTC TGAGAAGTTTGTCTT




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-60-
GAGAAGTTTGTCTTG
AGAAGTTTGTCTTGC
GAAGTTTGTCTTGCA
AAGTTTGTCTTGCAA
AGTTTGTCTTGCAAT
GTTTGTCTTGCAATG
TTTGTCTTGCAATGT
TTGTCTTGCAATGTA
TGTCTTGCAATGTAT
GTCTTGCAATGTATT
TCTTGCAATGTATTT
CTTGCAATGTATTTA
TTGCAATGTATTTAT
TGCAATGTATTTATA
GCAATGTATTTATAA
CAATGTATTTATAAA
AATGTATTTATAAAT
ATGTATTTATAAATA
TGTATTTATAAATAG
GTATTTATAAATAGT
TATTTATAAATAGTA
ATTTATAAATAGTAA
TTTATAAATAGTAAA
TTATAAATAGTAAAT
TATAAATAGTAAATA
ATAAATAGTAAATAA
TAAATAGTAAATAAA
AAATAGTAAATAAAG
AATAGTAAATAAAGT
ATAGTAAATAAAGTT
TAGTAAATAAAGTTT
AGTAAATAAAGTTTT
GTAAATAAAGTTTTT
TAAATAAAGTTTTTA
AAATAAAGTTTTTAC
AATAAAGTTTTTACC
ATAAAGTTTTTACCA
TAAAGTTTTTACCAT
AAAGTTTTTACCATT
EXAMPLE 8
Antisense oligonucleotides to IGF-I may be selected from molecules capable of
interacting with
one or more of the following sense oligonucleotides:
TTTTTTTTTTTTTTG TTTTTTTTTTTGAGA TTTTTTTTGAGAAAG
TTTTTTTTTTTTTGA TTTTTTTTTTGAGAA TTTTTTTGAGAAAGG
TTTTTTTTTTTTGAG TTTTTTTTTGAGAAA TTTTTTGAGAAAGGG


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-61-
TTTTTGAGAAAGGGA GGAGGAGGGTCCCCG CTCTCGCTCTGGCCG


TTTTGAGAAAGGGAA GAGGAGGGTCCCCGA TCTCGCTCTGGCCGA


TTTGAGAAAGGGAAT AGGAGGGTCCCCGAC CTCGCTCTGGCCGAC


TTGAGAAAGGGAATT GGAGGGTCCCCGACC TCGCTCTGGCCGACG


S TGAGAAAGGGAATTT GAGGGTCCCCGACCT CGCTCTGGCCGACGA


GAGAAAGGGAATTTC AGGGTCCCCGACCTC GCTCTGGCCGACGAG


AGAAAGGGAATTTCA GGGTCCCCGACCTCG CTCTGGCCGACGAGT


GAAAGGGAATTTCAT GGTCCCCGACCTCGC TCTGGCCGACGAGTG


AAAGGGAATTTCATC GTCCCCGACCTCGCT CTGGCCGACGAGTGG


10AAGGGAATTTCATCC TCCCCGACCTCGCTG TGGCCGACGAGTGGA


AGGGAATTTCATCCC CCCCGACCTCGCTGT GGCCGACGAGTGGAG


GGGAATTTCATCCCA CCCGACCTCGCTGTG GCCGACGAGTGGAGA


GGAATTTCATCCCAA CCGACCTCGCTGTGG CCGACGAGTGGAGAA


GAATTTCATCCCAAA CGACCTCGCTGTGGG CGACGAGTGGAGAAA


15AATTTCATCCCAAAT GACCTCGCTGTGGGG GACGAGTGGAGAAAT


ATTTCATCCCAAATA ACCTCGCTGTGGGGG ACGAGTGGAGAAATC


TTTCATCCCAAATAA CCTCGCTGTGGGGGC CGAGTGGAGAAATCT


TTCATCCCAAATAAA CTCGCTGTGGGGGCT GAGTGGAGAAATCTG


TCATCCCAAATAAAA TCGCTGTGGGGGCTC AGTGGAGAAATCTGC


20CATCCCAAATAAAAG CGCTGTGGGGGCTCC GTGGAGAAATCTGCG


ATCCCAAATAAAAGG GCTGTGGGGGCTCCT TGGAGAAATCTGCGG


TCCCAAATAAAAGGA CTGTGGGGGCTCCTG GGAGAAATCTGCGGG


CCCAAATAAAAGGAA TGTGGGGGCTCCTGT GAGAAATCTGCGGGC


CCAAATAAAAGGAAT GTGGGGGCTCCTGTT AGAAATCTGCGGGCC


25CAAATAAAAGGAAT'G TGGGGGCTCCTGTTT GAAATCTGCGGGCCA


AAATAAAAGGAATGA GGGGGCTCCTGTTTC AAATCTGCGGGCCAG


AATAAAAGGAATGAA GGGGCTCCTGTTTCT AATCTGCGGGCCAGG


ATAAAAGGAATGAAG GGGCTCCTGTTTCTC ATCTGCGGGCCAGGC


TAAAAGGAATGAAGT GGCTCCTGTTTCTCT TCTGCGGGCCAGGCA


30AAAAGGAATGAAGTC GCTCCTGTTTCTCTC CTGCGGGCCAGGCAT


AAAGGAATGAAGTCT CTCCTGTTTCTCTCC TGCGGGCCAGGCATC


AAGGAATGAAGTCTG TCCTGTTTCTCTCCG GCGGGCCAGGCATCG


AGGAATGAAGTCTGG CCTGTTTCTCTCCGC CGGGCCAGGCATCGA


GGAATGAAGTCTGGC CTGTTTCTCTCCGCC GGGCCAGGCATCGAC


35GAATGAAGTCTGGCT TGTTTCTCTCCGCCG GGCCAGGCATCGACA


AATGAAGTCTGGCTC GTTTCTCTCCGCCGC GCCAGGCATCGACAT


ATGAAGTCTGGCTCC TTTCTCTCCGCCGCG CCAGGCATCGACATC


TGAAGTCTGGCTCCG TTCTCTCCGCCGCGC CAGGCATCGACATCC


GAAGTCTGGCTCCGG TCTCTCCGCCGCGCT AGGCATCGACATCCG


40AAGTCTGGCTCCGGA CTCTCCGCCGCGCTC GGCATCGACATCCGC


AGTCTGGCTCCGGAG TCTCCGCCGCGCTCT GCATCGACATCCGCA


GTCTGGCTCCGGAGG CTCCGCCGCGCTCTC CATCGACATCCGCAA


TCTGGCTCCGGAGGA TCCGCCGCGCTCTCG ATCGACATCCGCAAC


CTGGCTCCGGAGGAG CCGCCGCGCTCTCGC TCGACATCCGCAACG


45TGGCTCCGGAGGAGG CGCCGCGCTCTCGCT CGACATCCGCAACGA


GGCTCCGGAGGAGGG GCCGCGCTCTCGCTC GACATCCGCAACGAC


GCTCCGGAGGAGGGT CCGCGCTCTCGCTCT ACATCCGCAACGACT


CTCCGGAGGAGGGTC CGCGCTCTCGCTCTG CATCCGCAACGACTA


TCCGGAGGAGGGTCC GCGCTCTCGCTCTGG ATCCGCAACGACTAT


50CCGGAGGAGGGTCCC CGCTCTCGCTCTGGC TCCGCAACGACTATC


CGGAGGAGGGTCCCC GCTCTCGCTCTGGCC CCGCAACGACTATCA




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-62-
CGCAACGACTATCAG GGCTACCTCCACATC CGCTTCCCCAAGCTC


GCAACGACTATCAGC GCTACCTCCACATCC GCTTCCCCAAGCTCA


CAACGACTATCAGCA CTACCTCCACATCCT CTTCCCCAAGCTCAC


AACGACTATCAGCAG TACCTCCACATCCTG TTCCCCAAGCTCACG


ACGACTATCAGCAGC ACCTCCACATCCTGC TCCCCAAGCTCACGG


CGACTATCAGCAGCT CCTCCACATCCTGCT CCCCAAGCTCACGGT


GACTATCAGCAGCTG CTCCACATCCTGCTC CCCAAGCTCACGGTC


ACTATCAGCAGCTGA TCCACATCCTGCTCA CCAAGCTCACGGTCA


CTATCAGCAGCTGAA CCACATCCTGCTCAT CAAGCTCACGGTCAT


10TATCAGCAGCTGAAG CACATCCTGCTCATC AAGCTCACGGTCATT


ATCAGCAGCTGAAGC ACATCCTGCTCATCT AGCTCACGGTCATTA


TCAGCAGCTGAAGCG CATCCTGCTCATCTC GCTCACGGTCATTAC


CAGCAGCTGAAGCGC ATCCTGCTCATCTCC CTCACGGTCATTACC


AGCAGCTGAAGCGCC TCCTGCTCATCTCCA TCACGGTCATTACCG


15GCAGCTGAAGCGCCT CCTGCTCATCTCCAA CACGGTCATTACCGA


CAGCTGAAGCGCCTG CTGCTCATCTCCAAG ACGGTCATTACCGAG


AGCTGAAGCGCCTGG TGCTCATCTCCAAGG CGGTCATTACCGAGT


GCTGAAGCGCCTGGA GCTCATCTCCAAGGC . GGTCATTACCGAGTA


CTGAAGCGCCTGGAG CTCATCTCCAAGGCC GTCATTACCGAGTAC


20TGAAGCGCCTGGAGA TCATCTCCAAGGCCG TCATTACCGAGTACT


GAAGCGCCTGGAGAA CATCTCCAAGGCCGA CATTACCGAGTACTT


AAGCGCCTGGAGAAC ATCTCCAAGGCCGAG ATTACCGAGTACTTG


AGCGCCTGGAGAACT TCTCCAAGGCCGAGG TTACCGAGTACTTGC


GCGCCTGGAGAACTG CTCCAAGGCCGAGGA TACCGAGTACTTGCT


25CGCCTGGAGAACTGC TCCAAGGCCGAGGAC ACCGAGTACTTGCTG


GCCTGGAGAACTGCA CCAAGGCCGAGGACT CCGAGTACTTGCTGC


CCTGGAGAACTGCAC CAAGGCCGAGGACTA CGAGTACTTGCTGCT


CTGGAGAACTGCACG AAGGCCGAGGACTAC GAGTACTTGCTGCTG


TGGAGAACTGCACGG AGGCCGAGGACTACC AGTACTTGCTGCTGT


30GGAGAACTGCACGGT GGCCGAGGACTACCG GTACTTGCTGCTGTT


GAGAACTGCACGGTG GCCGAGGACTACCGC TACTTGCTGCTGTTC


AGAACTGCACGGTGA CCGAGGACTACCGCA ACTTGCTGCTGTTCC


GAACTGCACGGTGAT CGAGGACTACCGCAG CTTGCTGCTGTTCCG


AACTGCACGGTGATC GAGGACTACCGCAGC TTGCTGCTGTTCCGA


35ACTGCACGGTGATCG AGGACTACCGCAGCT TGCTGCTGTTCCGAG


CTGCACGGTGATCGA GGACTACCGCAGCTA GCTGCTGTTCCGAGT


TGCACGGTGATCGAG GACTACCGCAGCTAC CTGCTGTTCCGAGTG


GCACGGTGATCGAGG ACTACCGCAGCTACC TGCTGTTCCGAGTGG


CACGGTGATCGAGGG CTACCGCAGCTACCG GCTGTTCCGAGTGGC


40ACGGTGATCGAGGGC TACCGCAGCTACCGC CTGTTCCGAGTGGCT


CGGTGATCGAGGGCT ACCGCAGCTACCGCT TGTTCCGAGTGGCTG


GGTGATCGAGGGCTA CCGCAGCTACCGCTT GTTCCGAGTGGCTGG


GTGATCGAGGGCTAC CGCAGCTACCGCTTC TTCCGAGTGGCTGGC


TGATCGAGGGCTACC GCAGCTACCGCTTCC TCCGAGTGGCTGGCC


45GATCGAGGGCTACCT CAGCTACCGCTTCCC CCGAGTGGCTGGCCT


ATCGAGGGCTACCTC AGCTACCGCTTCCCC CGAGTGGCTGGCCTC


TCGAGGGCTACCTCC GCTACCGCTTCCCCA GAGTGGCTGGCCTCG


CGAGGGCTACCTCCA CTACCGCTTCCCCAA AGTGGCTGGCCTCGA


GAGGGCTACCTCCAC TACCGCTTCCCCAAG GTGGCTGGCCTCGAG


50AGGGCTACCTCCACA ACCGCTTCCCCAAGC TGGCTGGCCTCGAGA


GGGCTACCTCCACAT CCGCTTCCCCAAGCT GGCTGGCCTCGAGAG




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-63-
GCTGGCCTCGAGAGC GGCTGGAAACTCTTC CTCAAGGATATTGGG


CTGGCCTCGAGAGCC GCTGGAAACTCTTCT TCAAGGATATTGGGC


TGGCCTCGAGAGCCT CTGGAAACTCTTCTA CAAGGATATTGGGCT


GGCCTCGAGAGCCTC TGGAAACTCTTCTAC AAGGATATTGGGCTT


GCCTCGAGAGCCTCG GGAAACTCTTCTACA AGGATATTGGGCTTT


CCTCGAGAGCCTCGG GAAACTCTTCTACAA GGATATTGGGCTTTA


CTCGAGAGCCTCGGA AAACTCTTCTACAAC GATATTGGGCTTTAC


TCGAGAGCCTCGGAG AACTCTTCTACAACT ATATTGGGCTTTACA


CGAGAGCCTCGGAGA ACTCTTCTACAACTA TATTGGGCTTTACAA


10GAGAGCCTCGGAGAC CTCTTCTACAACTAC ATTGGGCTTTACAAC


AGAGCCTCGGAGACC TCTTCTACAACTACG TTGGGCTTTACAACC


GAGCCTCGGAGACCT CTTCTACAACTACGC TGGGCTTTACAACCT


AGCCTCGGAGACCTC TTCTACAACTACGCC GGGCTTTACAACCTG


GCCTCGGAGACCTCT TCTACAACTACGCCC GGCTTTACAACCTGA


15CCTCGGAGACCTCTT CTACAACTACGCCCT GCTTTACAACCTGAG


CTCGGAGACCTCTTC TACAACTACGCCCTG CTTTACAACCTGAGG


TCGGAGACCTCTTCC ACAACTACGCCCTGG TTTACAACCTGAGGA


CGGAGACCTCTTCCC CAACTACGCCCTGGT TTACAACCTGAGGAA


GGAGACCTCTTCCCC AACTACGCCCTGGTC TACAACCTGAGGAAC


20GAGACCTCTTCCCCA ACTACGCCCTGGTCA ACAACCTGAGGAACA


AGACCTCTTCCCCAA CTACGCCCTGGTCAT CAACCTGAGGAACAT


GACCTCTTCCCCAAC TACGCCCTGGTCATC AACCTGAGGAACATT


ACCTCTTCCCCAACC ACGCCCTGGTCATCT ACCTGAGGAACATTA


CCTCTTCCCCAACCT CGCCCTGGTCATCTT CCTGAGGAACATTAC


25CTCTTCCCCAACCTC GCCCTGGTCATCTTC CTGAGGAACATTACT


TCTTCCCCAACCTCA CCCTGGTCATCTTCG TGAGGAACATTACTC


CTTCCCCAACCTCAC CCTGGTCATCTTCGA GAGGAACATTACTCG


TTCCCCAACCTCACG CTGGTCATCTTCGAG AGGAACATTACTCGG


TCCCCAACCTCACGG TGGTCATCTTCGAGA GGAACATTACTCGGG


30CCCCAACCTCACGGT GGTCATCTTCGAGAT GAACATTACTCGGGG


CCCAACCTCACGGTC GTCATCTTCGAGATG AACATTACTCGGGGG


CCAACCTCACGGTCA TCATCTTCGAGATGA ACATTACTCGGGGGG


CAACCTCACGGTCAT CATCTTCGAGATGAC CATTACTCGGGGGGC


AACCTCACGGTCATC ATCTTCGAGATGACC ATTACTCGGGGGGCC


35ACCTCACGGTCATCC TCTTCGAGATGACCA TTACTCGGGGGGCCA


CCTCACGGTCATCCG CTTCGAGATGACCAA TACTCGGGGGGCCAT


CTCACGGTCATCCGC TTCGAGATGACCAAT ACTCGGGGGGCCATC


TCACGGTCATCCGCG TCGAGATGACCAATC CTCGGGGGGCCATCA


CACGGTCATCCGCGG CGAGATGACCAATCT TCGGGGGGCCATCAG


40ACGGTCATCCGCGGC GAGATGACCAATCTC CGGGGGGCCATCAGG


CGGTCATCCGCGGCT AGATGACCAATCTCA GGGGGGCCATCAGGA


GGTCATCCGCGGCTG GATGACCAATCTCAA GGGGGCCATCAGGAT


GTCATCCGCGGCTGG ATGACCAATCTCAAG GGGGCCATCAGGATT


TCATCCGCGGCTGGA TGACCAATCTCAAGG GGGCCATCAGGATTG


45CATCCGCGGCTGGAA GACCAATCTCAAGGA GGCCATCAGGATTGA


ATCCGCGGCTGGAAA ACCAATCTCAAGGAT GCCATCAGGATTGAG


TCCGCGGCTGGAAAC CCAATCTCAAGGATA CCATCAGGATTGAGA


CCGCGGCTGGAAACT CAATCTCAAGGATAT CATCAGGATTGAGAA


CGCGGCTGGAAACTC AATCTCAAGGATATT ATCAGGATTGAGAAA


50GCGGCTGGAAACTCT ATCTCAAGGATATTG TCAGGATTGAGAAAA


CGGCTGGAAACTCTT TCTCAAGGATATTGG CAGGATTGAGAAAAA




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-64-
AGGATTGAGAAP~AAT CTGATCCTGGATGCG AAGGAATGTGGGGAC


GGATTGAGAA.P.AATG TGATCCTGGATGCGG AGGAATGTGGGGACC


GATTGAGAAAAATGC GATCCTGGATGCGGT GGAATGTGGGGACCT


ATTGAGAAAAATGCT ATCCTGGATGCGGTG GAATGTGGGGACCTG


S TTGAGAAAAATGCTG TCCTGGATGCGGTGT AATGTGGGGACCTGT


TGAGAAAAATGCTGA CCTGGATGCGGTGTC ATGTGGGGACCTGTG


GAGAAAAATGCTGAC CTGGATGCGGTGTCC TGTGGGGACCTGTGT


AGAAAAATGCTGACC TGGATGCGGTGTCCA GTGGGGACCTGTGTC


GAAAAATGCTGACCT GGATGCGGTGTCCAA TGGGGACCTGTGTCC


10AAAA.ATGCTGACCTC GATGCGGTGTCCAAT GGGGACCTGTGTCCA


AAAATGCTGACCTCT ATGCGGTGTCCAATA GGGACCTGTGTCCAG


AAATGCTGACCTCTG TGCGGTGTCCAATAA GGACCTGTGTCCAGG


AATGCTGACCTCTGT GCGGTGTCCAATAAC GACCTGTGTCCAGGG


ATGCTGACCTCTGTT CGGTGTCCAATAACT ACCTGTGTCCAGGGA


15TGCTGACCTCTGTTA GGTGTCCAATAACTA CCTGTGTCCAGGGAC


GCTGACCTCTGTTAC GTGTCCAATAACTAC CTGTGTCCAGGGACC


CTGACCTCTGTTACC TGTCCAATAACTACA TGTGTCCAGGGACCA


TGACCTCTGTTACCT GTCCAATAACTACAT GTGTCCAGGGACCAT


GACCTCTGTTACCTC TCCAATAACTACATT TGTCCAGGGACCATG


20ACCTCTGTTACCTCT CCAATAACTACATTG GTCCAGGGACCATGG


CCTCTGTTACCTCTC CAATAACTACATTGT TCCAGGGACCATGGA


CTCTGTTACCTCTCC AATAACTACATTGTG CCAGGGACCATGGAG


TCTGTTACCTCTCCA ATAACTACATTGTGG CAGGGACCATGGAGG


CTGTTACCTCTCCAC TAACTACATTGTGGG AGGGACCATGGAGGA


25TGTTACCTCTCCACT AACTACATTGTGGGG GGGACCATGGAGGAG


GTTACCTCTCCACTG ACTACATTGTGGGGA GGACCATGGAGGAGA


TTACCTCTCCACTGT CTACATTGTGGGGAA GACCATGGAGGAGAA


TACCTCTCCACTGTG TACATTGTGGGGAAT ACCATGGAGGAGAAG


ACCTCTCCACTGTGG ACATTGTGGGGAATA CCATGGAGGAGAAGC


30CCTCTCCACTGTGGA CATTGTGGGGAATAA CATGGAGGAGAAGCC


CTCTCCACTGTGGAC ATTGTGGGGAATAAG ATGGAGGAGAAGCCG


TCTCCACTGTGGACT TTGTGGGGAATAAGC TGGAGGAGAAGCCGA


CTCCACTGTGGACTG TGTGGGGAATAAGCC GGAGGAGAAGCCGAT


TCCACTGTGGACTGG GTGGGGAATAAGCCC GAGGAGAAGCCGATG


35CCACTGTGGACTGGT TGGGGAATAAGCCCC AGGAGAAGCCGATGT


CACTGTGGACTGGTC GGGGAATAAGCCCCC GGAGAAGCCGATGTG


ACTGTGGACTGGTCC GGGAATAAGCCCCCA GAGAAGCCGATGTGT


CTGTGGACTGGTCCC GGAATAAGCCCCCAA AGAAGCCGATGTGTG


TGTGGACTGGTCCCT GAATAAGCCCCCAAA GAAGCCGATGTGTGA


40GTGGACTGGTCCCTG AATAAGCCCCCAAAG AAGCCGATGTGTGAG


TGGACTGGTCCCTGA ATAAGCCCCCAAAGG AGCCGATGTGTGAGA


GGACTGGTCCCTGAT TAAGCCCCCAAAGGA GCCGATGTGTGAGAA


GACTGGTCCCTGATC AAGCCCCCAAAGGAA CCGATGTGTGAGAAG


ACTGGTCCCTGATCC AGCCCCCAAAGGAAT CGATGTGTGAGAAGA


45CTGGTCCCTGATCCT GCCCCCAAAGGAATG GATGTGTGAGAAGAC


TGGTCCCTGATCCTG CCCCCAAAGGAATGT ATGTGTGAGAAGACC


GGTCCCTGATCCTGG CCCCAAAGGAATGTG TGTGTGAGAAGACCA


GTCCCTGATCCTGGA CCCAAAGGAATGTGG GTGTGAGAAGACCAC


TCCCTGATCCTGGAT CCAAAGGAATGTGGG TGTGAGAAGACCACC


50CCCTGATCCTGGATG CAAAGGAATGTGGGG GTGAGAAGACCACCA


CCTGATCCTGGATGC AAAGGAATGTGGGGA TGAGAAGACCACCAT




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GAGAAGACCACCATC CGCTGCCAGAAAATG AACAATGAGTGCTGC


AGAAGACCACCATCA GCTGCCAGAAAATGT ACAATGAGTGCTGCC


GAAGACCACCATCAA CTGCCAGAAAATGTG CAATGAGTGCTGCCA


AAGACCACCATCAAC TGCCAGAAAATGTGC AATGAGTGCTGCCAC


AGACCACCATCAACA GCCAGAAAATGTGCC ATGAGTGCTGCCACC


GACCACCATCAACAA CCAGAAAATGTGCCC TGAGTGCTGCCACCC


ACCACCATCAACAAT CAGAAAATGTGCCCA GAGTGCTGCCACCCC


CCACCATCAACAATG AGAAAATGTGCCCAA AGTGCTGCCACCCCG


CACCATCAACAATGA GAAAATGTGCCCAAG GTGCTGCCACCCCGA


10ACCATCAACAATGAG AAAATGTGCCCAAGC TGCTGCCACCCCGAG


CCATCAACAATGAGT AAATGTGCCCAAGCA GCTGCCACCCCGAGT


CATCAACAATGAGTA AATGTGCCCAAGCAC CTGCCACCCCGAGTG


ATCAACAATGAGTAC ATGTGCCCAAGCACG TGCCACCCCGAGTGC


TCAACAATGAGTACA TGTGCCCAAGCACGT GCCACCCCGAGTGCC


15CAACAATGAGTACAA GTGCCCAAGCACGTG CCACCCCGAGTGCCT


AACAATGAGTACAAC TGCCCAAGCACGTGT CACCCCGAGTGCCTG


ACAATGAGTACAACT GCCCAAGCACGTGTG ACCCCGAGTGCCTGG


CAATGAGTACAACTA CCCAAGCACGTGTGG CCCCGAGTGCCTGGG


AATGAGTACAACTAC CCAAGCACGTGTGGG CCCGAGTGCCTGGGC


20ATGAGTACAACTACC CAAGCACGTGTGGGA CCGAGTGCCTGGGCA


TGAGTACAACTACCG AAGCACGTGTGGGAA CGAGTGCCTGGGCAG


GAGTACAACTACCGC AGCACGTGTGGGAAG GAGTGCCTGGGCAGC


AGTACAACTACCGCT GCACGTGTGGGAAGC AGTGCCTGGGCAGCT


GTACAACTACCGCTG CACGTGTGGGAAGCG GTGCCTGGGCAGCTG


25TACAACTACCGCTGC ACGTGTGGGAAGCGG TGCCTGGGCAGCTGC


ACAACTACCGCTGCT CGTGTGGGAAGCGGG GCCTGGGCAGCTGCA


CAACTACCGCTGCTG GTGTGGGAAGCGGGC CCTGGGCAGCTGCAG


AACTACCGCTGCTGG TGTGGGAAGCGGGCG CTGGGCAGCTGCAGC


ACTACCGCTGCTGGA GTGGGAAGCGGGCGT TGGGCAGCTGCAGCG


30CTACCGCTGCTGGAC TGGGAAGCGGGCGTG GGGCAGCTGCAGCGC


TACCGCTGCTGGACC GGGAAGCGGGCGTGC GGCAGCTGCAGCGCG


ACCGCTGCTGGACCA GGAAGCGGGCGTGCA GCAGCTGCAGCGCGC


CCGCTGCTGGACCAC GAAGCGGGCGTGCAC CAGCTGCAGCGCGCC


CGCTGCTGGACCACA AAGCGGGCGTGCACC AGCTGCAGCGCGCCT


35GCTGCTGGACCACAA AGCGGGCGTGCACCG GCTGCAGCGCGCCTG


CTGCTGGACCACAAA GCGGGCGTGCACCGA CTGCAGCGCGCCTGA


TGCTGGACCACAAAC CGGGCGTGCACCGAG TGCAGCGCGCCTGAC


GCTGGACCACAAACC GGGCGTGCACCGAGA GCAGCGCGCCTGACA


CTGGACCACAAACCG GGCGTGCACCGAGAA CAGCGCGCCTGACAA


40TGGACCACAAACCGC GCGTGCACCGAGAAC AGCGCGCCTGACAAC


GGACCACAAACCGCT CGTGCACCGAGAACA GCGCGCCTGACAACG


GACCACAAACCGCTG GTGCACCGAGAACAA CGCGCCTGACAACGA


ACCACAAACCGCTGC TGCACCGAGAACAAT GCGCCTGACAACGAC


CCACAAACCGCTGCC GCACCGAGAACAATG CGCCTGACAACGACA


45CACAAACCGCTGCCA CACCGAGAACAATGA GCCTGACAACGACAC


ACAAACCGCTGCCAG ACCGAGAACAATGAG CCTGACAACGACACG


CAAACCGCTGCCAGA CCGAGAACAATGAGT CTGACAACGACACGG


AAACCGCTGCCAGAA CGAGAACAATGAGTG TGACAACGACACGGC


AACCGCTGCCAGAAA GAGAACAATGAGTGC GACAACGACACGGCC


50ACCGCTGCCAGAAAA AGAACAATGAGTGCT ACAACGACACGGCCT


CCGCTGCCAGAAAAT GAACAATGAGTGCTG CAACGACACGGCCTG




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AACGACACGGCCTGT GTGCCTGCCTGCCCG GACCGTGACTTCTGC


ACGACACGGCCTGTG TGCCTGCCTGCCCGC ACCGTGACTTCTGCG


CGACACGGCCTGTGT GCCTGCCTGCCCGCC CCGTGACTTCTGCGC


GACACGGCCTGTGTA CCTGCCTGCCCGCCC CGTGACTTCTGCGCC


ACACGGCCTGTGTAG CTGCCTGCCCGCCCA GTGACTTCTGCGCCA


CACGGCCTGTGTAGC TGCCTGCCCGCCCAA TGACTTCTGCGCCAA


ACGGCCTGTGTAGCT GCCTGCCCGCCCAAC GACTTCTGCGCCAAC


CGGCCTGTGTAGCTT CCTGCCCGCCCAACA ACTTCTGCGCCAACA


GGCCTGTGTAGCTTG CTGCCCGCCCAACAC CTTCTGCGCCAACAT


10GCCTGTGTAGCTTGC TGCCCGCCCAACACC TTCTGCGCCAACATC


CCTGTGTAGCTTGCC GCCCGCCCAACACCT TCTGCGCCAACATCC


CTGTGTAGCTTGCCG CCCGCCCAACACCTA CTGCGCCAACATCCT


TGTGTAGCTTGCCGC CCGCCCAACACCTAC TGCGCCAACATCCTC


GTGTAGCTTGCCGCC CGCCCAACACCTACA GCGCCAACATCCTCA


15TGTAGCTTGCCGCCA GCCCAACACCTACAG CGCCAACATCCTCAG


GTAGCTTGCCGCCAC CCCAACACCTACAGG GCCAACATCCTCAGC


TAGCTTGCCGCCACT CCAACACCTACAGGT CCAACATCCTCAGCG


AGCTTGCCGCCACTA CAACACCTACAGGTT CAACATCCTCAGCGC


GCTTGCCGCCACTAC AACACCTACAGGTTT AACATCCTCAGCGCC


20CTTGCCGCCACTACT ACACCTACAGGTTTG ACATCCTCAGCGCCG


TTGCCGCCACTACTA CACCTACAGGTTTGA CATCCTCAGCGCCGA


TGCCGCCACTACTAC ACCTACAGGTTTGAG ATCCTCAGCGCCGAG


GCCGCCACTACTACT CCTACAGGTTTGAGG TCCTCAGCGCCGAGA


CCGCCACTACTACTA CTACAGGTTTGAGGG CCTCAGCGCCGAGAG


25CGCCACTACTACTAT TACAGGTTTGAGGGC CTCAGCGCCGAGAGC


GCCACTACTACTATG ACAGGTTTGAGGGCT TCAGCGCCGAGAGCA


CCACTACTACTATGC CAGGTTTGAGGGCTG CAGCGCCGAGAGCAG


CACTACTACTATGCC AGGTTTGAGGGCTGG AGCGCCGAGAGCAGC


ACTACTACTATGCCG GGTTTGAGGGCTGGC GCGCCGAGAGCAGCG


30CTACTACTATGCCGG GTTTGAGGGCTGGCG CGCCGAGAGCAGCGA


TACTACTATGCCGGT TTTGAGGGCTGGCGC GCCGAGAGCAGCGAC


ACTACTATGCCGGTG TTGAGGGCTGGCGCT CCGAGAGCAGCGACT


CTACTATGCCGGTGT TGAGGGCTGGCGCTG CGAGAGCAGCGACTC


TACTATGCCGGTGTC GAGGGCTGGCGCTGT GAGAGCAGCGACTCC


35ACTATGCCGGTGTCT AGGGCTGGCGCTGTG AGAGCAGCGACTCCG


CTATGCCGGTGTCTG GGGCTGGCGCTGTGT GAGCAGCGACTCCGA


TATGCCGGTGTCTGT GGCTGGCGCTGTGTG AGCAGCGACTCCGAG


ATGCCGGTGTCTGTG GCTGGCGCTGTGTGG GCAGCGACTCCGAGG


TGCCGGTGTCTGTGT CTGGCGCTGTGTGGA CAGCGACTCCGAGGG


40GCCGGTGTCTGTGTG TGGCGCTGTGTGGAC AGCGACTCCGAGGGG


CCGGTGTCTGTGTGC GGCGCTGTGTGGACC GCGACTCCGAGGGGT


CGGTGTCTGTGTGCC GCGCTGTGTGGACCG CGACTCCGAGGGGTT


GGTGTCTGTGTGCCT CGCTGTGTGGACCGT GACTCCGAGGGGTTT


GTGTCTGTGTGCCTG GCTGTGTGGACCGTG ACTCCGAGGGGTTTG


45TGTCTGTGTGCCTGC CTGTGTGGACCGTGA CTCCGAGGGGTTTGT


GTCTGTGTGCCTGCC TGTGTGGACCGTGAC TCCGAGGGGTTTGTG


TCTGTGTGCCTGCCT GTGTGGACCGTGACT CCGAGGGGTTTGTGA


CTGTGTGCCTGCCTG TGTGGACCGTGACTT CGAGGGGTTTGTGAT


TGTGTGCCTGCCTGC GTGGACCGTGACTTC GAGGGGTTTGTGATC


50GTGTGCCTGCCTGCC TGGACCGTGACTTCT AGGGGTTTGTGATCC


TGTGCCTGCCTGCCC GGACCGTGACTTCTG GGGGTTTGTGATCCA




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GGGTTTGTGATCCAC ATCCGCAACGGCAGC CCGAAGGTCTGTGAG


GGTTTGTGATCCACG TCCGCAACGGCAGCC CGAAGGTCTGTGAGG


GTTTGTGATCCACGA CCGCAACGGCAGCCA GAAGGTCTGTGAGGA


TTTGTGATCCACGAC CGCAACGGCAGCCAG AAGGTCTGTGAGGAA


TTGTGATCCACGACG GCAACGGCAGCCAGA AGGTCTGTGAGGAAG


TGTGATCCACGACGG CAACGGCAGCCAGAG GGTCTGTGAGGAAGA


GTGATCCACGACGGC AACGGCAGCCAGAGC GTCTGTGAGGAAGAA


TGATCCACGACGGCG ACGGCAGCCAGAGCA TCTGTGAGGAAGAAA


GATCCACGACGGCGA CGGCAGCCAGAGCAT CTGTGAGGAAGAAAA


10ATCCACGACGGCGAG GGCAGCCAGAGCATG TGTGAGGAAGAAAAG


TCCACGACGGCGAGT GCAGCCAGAGCATGT GTGAGGAAGAAAAGA


CCACGACGGCGAGTG CAGCCAGAGCATGTA TGAGGAAGAAAAGAA


CACGACGGCGAGTGC AGCCAGAGCATGTAC GAGGAAGAAAAGAAA


ACGACGGCGAGTGCA GCCAGAGCATGTACT AGGAAGAAAAGAAAA


15CGACGGCGAGTGCAT CCAGAGCATGTACTG GGAAGAAAAGAAAAC


GACGGCGAGTGCATG CAGAGCATGTACTGC GAAGAAAAGAAAACA


ACGGCGAGTGCATGC AGAGCATGTACTGCA AAGAAAAGAAAACAA


CGGCGAGTGCATGCA GAGCATGTACTGCAT AGAAAAGAAAACAAA


GGCGAGTGCATGCAG AGCATGTACTGCATC GAAAAGAAAACAAAG


20GCGAGTGCATGCAGG GCATGTACTGCATCC AAAAGAAAACAAAGA


CGAGTGCATGCAGGA CATGTACTGCATCCC AAAGAAAACAAAGAC


GAGTGCATGCAGGAG ATGTACTGCATCCCT AAGAAAACAAAGACC


AGTGCATGCAGGAGT TGTACTGCATCCCTT AGAAAACAAAGACCA


GTGCATGCAGGAGTG GTACTGCATCCCTTG GAAAACAAAGACCAT


25TGCATGCAGGAGTGC TACTGCATCCCTTGT AAAACAAAGACCATT


GCATGCAGGAGTGCC ACTGCATCCCTTGTG AAACAAAGACCATTG


CATGCAGGAGTGCCC CTGCATCCCTTGTGA AACAAAGACCATTGA


ATGCAGGAGTGCCCC TGCATCCCTTGTGAA ACAAAGACCATTGAT


TGCAGGAGTGCCCCT GCATCCCTTGTGAAG CAAAGACCATTGATT


30GCAGGAGTGCCCCTC CATCCCTTGTGAAGG AAAGACCATTGATTC


CAGGAGTGCCCCTCG ATCCCTTGTGAAGGT AAGACCATTGATTCT


AGGAGTGCCCCTCGG TCCCTTGTGAAGGTC AGACCATTGATTCTG


GGAGTGCCCCTCGGG CCCTTGTGAAGGTCC GACCATTGATTCTGT


GAGTGCCCCTCGGGC CCTTGTGAAGGTCCT ACCATTGATTCTGTT


35AGTGCCCCTCGGGCT CTTGTGAAGGTCCTT CCATTGATTCTGTTA


GTGCCCCTCGGGCTT TTGTGAAGGTCCTTG CATTGATTCTGTTAC


TGCCCCTCGGGCTTC TGTGAAGGTCCTTGC ATTGATTCTGTTACT


GCCCCTCGGGCTTCA GTGAAGGTCCTTGCC TTGATTCTGTTACTT


CCCCTCGGGCTTCAT TGAAGGTCCTTGCCC TGATTCTGTTACTTC


40CCCTCGGGCTTCATC GAAGGTCCTTGCCCG GATTCTGTTACTTCT


CCTCGGGCTTCATCC AAGGTCCTTGCCCGA ATTCTGTTACTTCTG


CTCGGGCTTCATCCG AGGTCCTTGCCCGAA TTCTGTTACTTCTGC


TCGGGCTTCATCCGC GGTCCTTGCCCGAAG TCTGTTACTTCTGCT


CGGGCTTCATCCGCA GTCCTTGCCCGAAGG CTGTTACTTCTGCTC


45GGGCTTCATCCGCAA TCCTTGCCCGAAGGT TGTTACTTCTGCTCA


GGCTTCATCCGCAAC CCTTGCCCGAAGGTC GTTACTTCTGCTCAG


GCTTCATCCGCAACG CTTGCCCGAAGGTCT TTACTTCTGCTCAGA


CTTCATCCGCAACGG TTGCCCGAAGGTCTG TACTTCTGCTCAGAT


TTCATCCGCAACGGC TGCCCGAAGGTCTGT ACTTCTGCTCAGATG


50TCATCCGCAACGGCA GCCCGAAGGTCTGTG CTTCTGCTCAGATGC


CATCCGCAACGGCAG CCCGAAGGTCTGTGA TTCTGCTCAGATGCT




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TCTGCTCAGATGCTC AACATCCGACGGGGG CTCATCGAGGTGGTG


CTGCTCAGATGCTCC ACATCCGACGGGGGA TCATCGAGGTGGTGA


TGCTCAGATGCTCCA CATCCGACGGGGGAA CATCGAGGTGGTGAC


GCTCAGATGCTCCAA ATCCGACGGGGGAAT ATCGAGGTGGTGACG


CTCAGATGCTCCAAG TCCGACGGGGGAATA TCGAGGTGGTGACGG


TCAGATGCTCCAAGG CCGACGGGGGAATAA CGAGGTGGTGACGGG


CAGATGCTCCAAGGA CGACGGGGGAATAAC GAGGTGGTGACGGGC


AGATGCTCCAAGGAT GACGGGGGAATAACA AGGTGGTGACGGGCT


GATGCTCCAAGGATG ACGGGGGAATAACAT GGTGGTGACGGGCTA


10ATGCTCCAAGGATGC CGGGGGAATAACATT GTGGTGACGGGCTAC


TGCTCCAAGGATGCA GGGGGAATAACATTG TGGTGACGGGCTACG


GCTCCAAGGATGCAC GGGGAATAACATTGC GGTGACGGGCTACGT


CTCCAAGGATGCACC GGGAATAACATTGCT GTGACGGGCTACGTG


TCCAAGGATGCACCA GGAATAACATTGCTT TGACGGGCTACGTGA


15CCAAGGATGCACCAT GAATAACATTGCTTC GACGGGCTACGTGAA


CAAGGATGCACCATC AATAACATTGCTTCA ACGGGCTACGTGAAG


AAGGATGCACCATCT ATAACATTGCTTCAG CGGGCTACGTGAAGA


AGGATGCACCATCTT TAACATTGCTTCAGA GGGCTACGTGAAGAT


GGATGCACCATCTTC AACATTGCTTCAGAG GGCTACGTGAAGATC


20GATGCACCATCTTCA ACATTGCTTCAGAGC GCTACGTGAAGATCC


ATGCACCATCTTCAA CATTGCTTCAGAGCT CTACGTGAAGATCCG


TGCACCATCTTCAAG ATTGCTTCAGAGCTG TACGTGAAGATCCGC


GCACCATCTTCAAGG TTGCTTCAGAGCTGG ACGTGAAGATCCGCC


CACCATCTTCAAGGG TGCTTCAGAGCTGGA CGTGAAGATCCGCCA


25ACCATCTTCAAGGGC GCTTCAGAGCTGGAG GTGAAGATCCGCCAT


CCATCTTCAAGGGCA CTTCAGAGCTGGAGA TGAAGATCCGCCATT


CATCTTCAAGGGCAA TTCAGAGCTGGAGAA GAAGATCCGCCATTC


ATCTTCAAGGGCAAT TCAGAGCTGGAGAAC AAGATCCGCCATTCT


TCTTCAAGGGCAATT CAGAGCTGGAGAACT AGATCCGCCATTCTC


30CTTCAAGGGCAATTT AGAGCTGGAGAACTT GATCCGCCATTCTCA


TTCAAGGGCAATTTG GAGCTGGAGAACTTC ATCCGCCATTCTCAT


TCAAGGGCAATTTGC AGCTGGAGAACTTCA TCCGCCATTCTCATG


CAAGGGCAATTTGCT GCTGGAGAACTTCAT CCGCCATTCTCATGC


AAGGGCAATTTGCTC CTGGAGAACTTCATG CGCCATTCTCATGCC


35AGGGCAATTTGCTCA TGGAGAACTTCATGG GCCATTCTCATGCCT


GGGCAATTTGCTCAT GGAGAACTTCATGGG CCATTCTCATGCCTT


GGCAATTTGCTCATT GAGAACTTCATGGGG CATTCTCATGCCTTG


GCAATTTGCTCATTA AGAACTTCATGGGGC ATTCTCATGCCTTGG


CAATTTGCTCATTAA GAACTTCATGGGGCT TTCTCATGCCTTGGT


40AATTTGCTCATTAAC AACTTCATGGGGCTC TCTCATGCCTTGGTC


ATTTGCTCATTAACA ACTTCATGGGGCTCA CTCATGCCTTGGTCT


TTTGCTCATTAACAT CTTCATGGGGCTCAT TCATGCCTTGGTCTC


TTGCTCATTAACATC TTCATGGGGCTCATC CATGCCTTGGTCTCC


TGCTCATTAACATCC TCATGGGGCTCATCG ATGCCTTGGTCTCCT


45GCTCATTAACATCCG CATGGGGCTCATCGA TGCCTTGGTCTCCTT


CTCATTAACATCCGA ATGGGGCTCATCGAG GCCTTGGTCTCCTTG


TCATTAACATCCGAC TGGGGCTCATCGAGG CCTTGGTCTCCTTGT


CATTAACATCCGACG GGGGCTCATCGAGGT CTTGGTCTCCTTGTC


ATTAACATCCGACGG GGGCTCATCGAGGTG TTGGTCTCCTTGTCC


50TTAACATCCGACGGG GGCTCATCGAGGTGG TGGTCTCCTTGTCCT


TAACATCCGACGGGG GCTCATCGAGGTGGT GGTCTCCTTGTCCTT




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GTCTCCTTGTCCTTC CTAGAAGGGAATTAC CTGTGGGACTGGGAC


TCTCCTTGTCCTTCC TAGAAGGGAATTACT TGTGGGACTGGGACC


CTCCTTGTCCTTCCT AGAAGGGAATTACTC GTGGGACTGGGACCA


TCCTTGTCCTTCCTA GAAGGGAATTACTCC TGGGACTGGGACCAC


CCTTGTCCTTCCTAA AAGGGAATTACTCCT GGGACTGGGACCACC


CTTGTCCTTCCTAAA AGGGAATTACTCCTT GGACTGGGACCACCG


TTGTCCTTCCTAAAA GGGAATTACTCCTTC GACTGGGACCACCGC


TGTCCTTCCTAAAAA GGAATTACTCCTTCT ACTGGGACCACCGCA


GTCCTTCCTAAAAAA GAATTACTCCTTCTA CTGGGACCACCGCAA


10TCCTTCCTAAAP~AAC AATTACTCCTTCTAC TGGGACCACCGCAAC


CCTTCCTAAAAAACC ATTACTCCTTCTACG GGGACCACCGCAACC


CTTCCTAAAAAACCT TTACTCCTTCTACGT GGACCACCGCAACCT


TTCCTAAAAAACCTT TACTCCTTCTACGTC GACCACCGCAACCTG


TCCTAAAAAACCTTC ACTCCTTCTACGTCC ACCACCGCAACCTGA


15CCTAAP.A.A.ACCTTCG CTCCTTCTACGTCCT CCACCGCAACCTGAC


CTP.AAP.AACCTTCGC TCCTTCTACGTCCTC CACCGCAACCTGACC


TAA.A.A.A.ACCTTCGCCCCTTCTACGTCCTCG ACCGCAACCTGACCA


AAAAAACCTTCGCCT CTTCTACGTCCTCGA CCGCAACCTGACCAT


P.AAAACCTTCGCCTC TTCTACGTCCTCGAC CGCAACCTGACCATC


20AAAACCTTCGCCTCA TCTACGTCCTCGACA GCAACCTGACCATCA


AAACCTTCGCCTCAT CTACGTCCTCGACAA CAACCTGACCATCAA


AACCTTCGCCTCATC TACGTCCTCGACAAC AACCTGACCATCAAA


ACCTTCGCCTCATCC ACGTCCTCGACAACC ACCTGACCATCAAAG


CCTTCGCCTCATCCT CGTCCTCGACAACCA CCTGACCATCAAAGC


25CTTCGCCTCATCCTA GTCCTCGACAACCAG CTGACCATCAAAGCA


TTCGCCTCATCCTAG TCCTCGACAACCAGA TGACCATCAAAGCAG


TCGCCTCATCCTAGG CCTCGACAACCAGAA GACCATCAAAGCAGG


CGCCTCATCCTAGGA CTCGACAACCAGAAC ACCATCAAAGCAGGG


GCCTCATCCTAGGAG TCGACAACCAGAACT CCATCAAAGCAGGGA


30CCTCATCCTAGGAGA CGACAACCAGAACTT CATCAAAGCAGGGAA


CTCATCCTAGGAGAG GACAACCAGAACTTG ATCAAAGCAGGGAAA


TCATCCTAGGAGAGG ACAACCAGAACTTGC TCAAAGCAGGGAAAA


CATCCTAGGAGAGGA CAACCAGAACTTGCA CAAAGCAGGGAAAAT


ATCCTAGGAGAGGAG AACCAGAACTTGCAG AAAGCAGGGAAAATG


35TCCTAGGAGAGGAGC ACCAGAACTTGCAGC AAGCAGGGAAAATGT


CCTAGGAGAGGAGCA CCAGAACTTGCAGCA AGCAGGGAAAATGTA


CTAGGAGAGGAGCAG CAGAACTTGCAGCAA GCAGGGAAAATGTAC


TAGGAGAGGAGCAGC AGAACTTGCAGCAAC CAGGGAAAATGTACT


AGGAGAGGAGCAGCT GAACTTGCAGCAACT AGGGAAAATGTACTT


40GGAGAGGAGCAGCTA AACTTGCAGCAACTG GGGAAAATGTACTTT


GAGAGGAGCAGCTAG ACTTGCAGCAACTGT GGAAAATGTACTTTG


AGAGGAGCAGCTAGA CTTGCAGCAACTGTG GAAAATGTACTTTGC


GAGGAGCAGCTAGAA TTGCAGCAACTGTGG AAAATGTACTTTGCT


AGGAGCAGCTAGAAG TGCAGCAACTGTGGG AAATGTACTTTGCTT


45GGAGCAGCTAGAAGG GCAGCAACTGTGGGA AATGTACTTTGCTTT


GAGCAGCTAGAAGGG CAGCAACTGTGGGAC ATGTACTTTGCTTTC


AGCAGCTAGAAGGGA AGCAACTGTGGGACT TGTACTTTGCTTTCA


GCAGCTAGAAGGGAA GCAACTGTGGGACTG GTACTTTGCTTTCAA


CAGCTAGAAGGGAAT CAACTGTGGGACTGG TACTTTGCTTTCAAT


50AGCTAGAAGGGAATT AACTGTGGGACTGGG ACTTTGCTTTCAATC


GCTAGAAGGGAATTA ACTGTGGGACTGGGA CTTTGCTTTCAATCC




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TTTGCTTTCAATCCC GTGACGGGGACTAAA AACGGGGAGAGAGCC


TTGCTTTCAATCCCA TGACGGGGACTAAAG ACGGGGAGAGAGCCT


TGCTTTCAATCCCAA GACGGGGACTAAAGG CGGGGAGAGAGCCTC


GCTTTCAATCCCAAA ACGGGGACTAAAGGG GGGGAGAGAGCCTCC


CTTTCAATCCCAAAT CGGGGACTAAAGGGC GGGAGAGAGCCTCCT


TTTCAATCCCAAATT GGGGACTAAAGGGCG GGAGAGAGCCTCCTG


TTCAATCCCAAATTA GGGACTAAAGGGCGC GAGAGAGCCTCCTGT


TCAATCCCAAATTAT GGACTAAAGGGCGCC AGAGAGCCTCCTGTG


CAATCCCAAATTATG GACTAAAGGGCGCCA GAGAGCCTCCTGTGA


10AATCCCAAATTATGT ACTAAAGGGCGCCAA AGAGCCTCCTGTGAA


ATCCCAAATTATGTG CTAAAGGGCGCCAAA GAGCCTCCTGTGAAA


TCCCAAATTATGTGT TAAAGGGCGCCAAAG AGCCTCCTGTGAAAG


CCCAAATTATGTGTT AAAGGGCGCCAAAGC GCCTCCTGTGAAAGT


CCAAATTATGTGTTT AAGGGCGCCAAAGCA CCTCCTGTGAAAGTG


15CAAATTATGTGTTTC AGGGCGCCAAAGCAA CTCCTGTGAAAGTGA


AAATTATGTGTTTCC GGGCGCCAAAGCAAA TCCTGTGAAAGTGAC


AATTATGTGTTTCCG GGCGCCAAAGCAAAG CCTGTGAAAGTGACG


ATTATGTGTTTCCGA GCGCCAAAGCAAAGG CTGTGAAAGTGACGT


TTATGTGTTTCCGAA CGCCAAAGCAAAGGG TGTGAAAGTGACGTC


20TATGTGTTTCCGAAA GCCAAAGCAAAGGGG GTGAAAGTGACGTCC


ATGTGTTTCCGAAAT CCAAAGCAAAGGGGA TGAAAGTGACGTCCT


TGTGTTTCCGAAATT CAAAGCAAAGGGGAC GAAAGTGACGTCCTG


GTGTTTCCGAAATTT AAAGCAAAGGGGACA AAAGTGACGTCCTGC


TGTTTCCGAAATTTA AAGCAAAGGGGACAT AAGTGACGTCCTGCA


25GTTTCCGAAATTTAC AGCAAAGGGGACATA AGTGACGTCCTGCAT


TTTCCGAAATTTACC GCAAAGGGGACATAA GTGACGTCCTGCATT


TTCCGAAATTTACCG CAAAGGGGACATAAA TGACGTCCTGCATTT


TCCGAAATTTACCGC AAAGGGGACATAAAC GACGTCCTGCATTTC


CCGAAATTTACCGCA AAGGGGACATAAACA ACGTCCTGCATTTCA


30CGAAATTTACCGCAT AGGGGACATAAACAC CGTCCTGCATTTCAC


GAAATTTACCGCATG GGGGACATAAACACC GTCCTGCATTTCACC


AAATTTACCGCATGG GGGACATAAACACCA TCCTGCATTTCACCT


AATTTACCGCATGGA GGACATAAACACCAG CCTGCATTTCACCTC


ATTTACCGCATGGAG GACATAAACACCAGG CTGCATTTCACCTCC


35TTTACCGCATGGAGG ACATAAACACCAGGA TGCATTTCACCTCCA


TTACCGCATGGAGGA CATAAACACCAGGAA GCATTTCACCTCCAC


TACCGCATGGAGGAA ATAAACACCAGGAAC CATTTCACCTCCACC


ACCGCATGGAGGAAG TAAACACCAGGAACA ATTTCACCTCCACCA


CCGCATGGAGGAAGT AAACACCAGGAACAA TTTCACCTCCACCAC


40CGCATGGAGGAAGTG AACACCAGGAACAAC TTCACCTCCACCACC


GCATGGAGGAAGTGA ACACCAGGAACAACG TCACCTCCACCACCA


CATGGAGGAAGTGAC CACCAGGAACAACGG CACCTCCACCACCAC


ATGGAGGAAGTGACG ACCAGGAACAACGGG ACCTCCACCACCACG


TGGAGGAAGTGACGG CCAGGAACAACGGGG CCTCCACCACCACGT


45GGAGGAAGTGACGGG CAGGAACAACGGGGA CTCCACCACCACGTC


GAGGAAGTGACGGGG AGGAACAACGGGGAG TCCACCACCACGTCG


AGGAAGTGACGGGGA GGAACAACGGGGAGA CCACCACCACGTCGA


GGAAGTGACGGGGAC GAACAACGGGGAGAG CACCACCACGTCGAA


GAAGTGACGGGGACT AACAACGGGGAGAGA ACCACCACGTCGAAG


50AAGTGACGGGGACTA ACAACGGGGAGAGAG CCACCACGTCGAAGA


AGTGACGGGGACTAA CAACGGGGAGAGAGC CACCACGTCGAAGAA




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ACCACGTCGAAGAAT GACTACAGGGATCTC AAGAATGTCACAGAG


CCACGTCGAAGAATC ACTACAGGGATCTCA AGAATGTCACAGAGT


CACGTCGAAGAATCG CTACAGGGATCTCAT GAATGTCACAGAGTA


ACGTCGAAGAATCGC TACAGGGATCTCATC AATGTCACAGAGTAT


CGTCGAAGAATCGCA ACAGGGATCTCATCA ATGTCACAGAGTATG


GTCGAAGAATCGCAT CAGGGATCTCATCAG TGTCACAGAGTATGA


TCGAAGAATCGCATC AGGGATCTCATCAGC GTCACAGAGTATGAT


CGAAGAATCGCATCA GGGATCTCATCAGCT TCACAGAGTATGATG


GAAGAATCGCATCAT GGATCTCATCAGCTT CACAGAGTATGATGG


10AAGAATCGCATCATC GATCTCATCAGCTTC ACAGAGTATGATGGG


AGAATCGCATCATCA ATCTCATCAGCTTCA CAGAGTATGATGGGC


GAATCGCATCATCAT TCTCATCAGCTTCAC AGAGTATGATGGGCA


AATCGCATCATCATA CTCATCAGCTTCACC GAGTATGATGGGCAG


ATCGCATCATCATAA TCATCAGCTTCACCG AGTATGATGGGCAGG


15TCGCATCATCATAAC CATCAGCTTCACCGT GTATGATGGGCAGGA


CGCATCATCATAACC ATCAGCTTCACCGTT TATGATGGGCAGGAT


GCATCATCATAACCT TCAGCTTCACCGTTT ATGATGGGCAGGATG


CATCATCATAACCTG CAGCTTCACCGTTTA TGATGGGCAGGATGC


ATCATCATAACCTGG AGCTTCACCGTTTAC GATGGGCAGGATGCC


20TCATCATAACCTGGC GCTTCACCGTTTACT ATGGGCAGGATGCCT


CATCATAACCTGGCA CTTCACCGTTTACTA TGGGCAGGATGCCTG


ATCATAACCTGGCAC TTCACCGTTTACTAC GGGCAGGATGCCTGC


TCATAACCTGGCACC TCACCGTTTACTACA GGCAGGATGCCTGCG


CATAACCTGGCACCG CACCGTTTACTACAA GCAGGATGCCTGCGG


25ATAACCTGGCACCGG ACCGTTTACTACAAG CAGGATGCCTGCGGC


TAACCTGGCACCGGT CCGTTTACTACAAGG AGGATGCCTGCGGCT


AACCTGGCACCGGTA CGTTTACTACAAGGA GGATGCCTGCGGCTC


ACCTGGCACCGGTAC GTTTACTACAAGGAA GATGCCTGCGGCTCC


CCTGGCACCGGTACC TTTACTACAAGGAAG ATGCCTGCGGCTCCA


30CTGGCACCGGTACCG TTACTACAAGGAAGC TGCCTGCGGCTCCAA


TGGCACCGGTACCGG TACTACAAGGAAGCA GCCTGCGGCTCCAAC


GGCACCGGTACCGGC ACTACAAGGAAGCAC CCTGCGGCTCCAACA


GCACCGGTACCGGCC CTACAAGGAAGCACC CTGCGGCTCCAACAG


CACCGGTACCGGCCC TACAAGGAAGCACCC TGCGGCTCCAACAGC


35ACCGGTACCGGCCCC ACAAGGAAGCACCCT GCGGCTCCAACAGCT


CCGGTACCGGCCCCC CAAGGAAGCACCCTT CGGCTCCAACAGCTG


CGGTACCGGCCCCCT AAGGAAGCACCCTTT GGCTCCAACAGCTGG


GGTACCGGCCCCCTG AGGAAGCACCCTTTA GCTCCAACAGCTGGA


GTACCGGCCCCCTGA GGAAGCACCCTTTAA CTCCAACAGCTGGAA


40TACCGGCCCCCTGAC GAAGCACCCTTTAAG TCCAACAGCTGGAAC


ACCGGCCCCCTGACT AAGCACCCTTTAAGA CCAACAGCTGGAACA


CCGGCCCCCTGACTA AGCACCCTTTAAGAA CAACAGCTGGAACAT


CGGCCCCCTGACTAC GCACCCTTTAAGAAT AACAGCTGGAACATG


GGCCCCCTGACTACA CACCCTTTAAGAATG ACAGCTGGAACATGG


45GCCCCCTGACTACAG ACCCTTTAAGAATGT CAGCTGGAACATGGT


CCCCCTGACTACAGG CCCTTTAAGAATGTC AGCTGGAACATGGTG


CCCCTGACTACAGGG CCTTTAAGAATGTCA GCTGGAACATGGTGG


CCCTGACTACAGGGA CTTTAAGAATGTCAC CTGGAACATGGTGGA


CCTGACTACAGGGAT TTTAAGAATGTCACA TGGAACATGGTGGAC


50CTGACTACAGGGATC TTAAGAATGTCACAG GGAACATGGTGGACG


TGACTACAGGGATCT TAAGAATGTCACAGA GAACATGGTGGACGT




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AACATGGTGGACGTG TTACTACATGGGCTG GTGACCCTCACCATG


ACATGGTGGACGTGG TACTACATGGGCTGA TGACCCTCACCATGG


CATGGTGGACGTGGA ACTACATGGGCTGAA GACCCTCACCATGGT


ATGGTGGACGTGGAC CTACATGGGCTGAAG ACCCTCACCATGGTG


TGGTGGACGTGGACC TACATGGGCTGAAGC CCCTCACCATGGTGG


GGTGGACGTGGACCT ACATGGGCTGAAGCC CCTCACCATGGTGGA


GTGGACGTGGACCTC CATGGGCTGAAGCCC CTCACCATGGTGGAG


TGGACGTGGACCTCC ATGGGCTGAAGCCCT TCACCATGGTGGAGA


GGACGTGGACCTCCC TGGGCTGAAGCCCTG CACCATGGTGGAGAA


10GACGTGGACCTCCCG GGGCTGAAGCCCTGG ACCATGGTGGAGAAC


ACGTGGACCTCCCGC GGCTGAAGCCCTGGA CCATGGTGGAGAACG


CGTGGACCTCCCGCC GCTGAAGCCCTGGAC CATGGTGGAGAACGA


GTGGACCTCCCGCCC CTGAAGCCCTGGACT ATGGTGGAGAACGAC


TGGACCTCCCGCCCA TGAAGCCCTGGACTC TGGTGGAGAACGACC


15GGACCTCCCGCCCAA GAAGCCCTGGACTCA GGTGGAGAACGACCA


GACCTCCCGCCCAAC AAGCCCTGGACTCAG GTGGAGAACGACCAT


ACCTCCCGCCCAACA AGCCCTGGACTCAGT TGGAGAACGACCATA


CCTCCCGCCCAACAA GCCCTGGACTCAGTA GGAGAACGACCATAT


CTCCCGCCCAACAAG CCCTGGACTCAGTAC GAGAACGACCATATC


20TCCCGCCCAACAAGG CCTGGACTCAGTACG AGAACGACCATATCC


CCCGCCCAACAAGGA CTGGACTCAGTACGC GAACGACCATATCCG


CCGCCCAACAAGGAC TGGACTCAGTACGCC AACGACCATATCCGT


CGCCCAACAAGGACG GGACTCAGTACGCCG ACGACCATATCCGTG


GCCCAACAAGGACGT GACTCAGTACGCCGT CGACCATATCCGTGG


25CCCAACAAGGACGTG ACTCAGTACGCCGTT GACCATATCCGTGGG


CCAACAAGGACGTGG CTCAGTACGCCGTTT ACCATATCCGTGGGG


CAACAAGGACGTGGA TCAGTACGCCGTTTA CCATATCCGTGGGGC


AACAAGGACGTGGAG CAGTACGCCGTTTAC CATATCCGTGGGGCC


ACAAGGACGTGGAGC AGTACGCCGTTTACG ATATCCGTGGGGCCA


30CAAGGACGTGGAGCC GTACGCCGTTTACGT TATCCGTGGGGCCAA


AAGGACGTGGAGCCC TACGCCGTTTACGTC ATCCGTGGGGCCAAG


AGGACGTGGAGCCCG ACGCCGTTTACGTCA TCCGTGGGGCCAAGA


GGACGTGGAGCCCGG CGCCGTTTACGTCAA CCGTGGGGCCAAGAG


GACGTGGAGCCCGGC GCCGTTTACGTCAAG CGTGGGGCCAAGAGT


35ACGTGGAGCCCGGCA CCGTTTACGTCAAGG GTGGGGCCAAGAGTG


CGTGGAGCCCGGCAT CGTTTACGTCAAGGC TGGGGCCAAGAGTGA


GTGGAGCCCGGCATC GTTTACGTCAAGGCT GGGGCCAAGAGTGAG


TGGAGCCCGGCATCT TTTACGTCAAGGCTG GGGCCAAGAGTGAGA


GGAGCCCGGCATCTT TTACGTCAAGGCTGT GGCCAAGAGTGAGAT


40GAGCCCGGCATCTTA TACGTCAAGGCTGTG GCCAAGAGTGAGATC


AGCCCGGCATCTTAC ACGTCAAGGCTGTGA CCAAGAGTGAGATCT


GCCCGGCATCTTACT CGTCAAGGCTGTGAC CAAGAGTGAGATCTT


CCCGGCATCTTACTA GTCAAGGCTGTGACC AAGAGTGAGATCTTG


CCGGCATCTTACTAC TCAAGGCTGTGACCC AGAGTGAGATCTTGT


45CGGCATCTTACTACA CAAGGCTGTGACCCT GAGTGAGATCTTGTA


GGCATCTTACTACAT AAGGCTGTGACCCTC AGTGAGATCTTGTAC


GCATCTTACTACATG AGGCTGTGACCCTCA GTGAGATCTTGTACA


CATCTTACTACATGG GGCTGTGACCCTCAC TGAGATCTTGTACAT


ATCTTACTACATGGG GCTGTGACCCTCACC GAGATCTTGTACATT


50TCTTACTACATGGGC CTGTGACCCTCACCA AGATCTTGTACATTC


CTTACTACATGGGCT TGTGACCCTCACCAT GATCTTGTACATTCG




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ATCTTGTACATTCGC CTTTCAGCATCGAAC TCTCTGCCCAACGGC


TCTTGTACATTCGCA TTTCAGCATCGAACT CTCTGCCCAACGGCA


CTTGTACATTCGCAC TTCAGCATCGAACTC TCTGCCCAACGGCAA


TTGTACATTCGCACC TCAGCATCGAACTCC CTGCCCAACGGCAAC


S TGTACATTCGCACCA CAGCATCGAACTCCT TGCCCAACGGCAACC


GTACATTCGCACCAA AGCATCGAACTCCTC GCCCAACGGCAACCT


TACATTCGCACCAAT GCATCGAACTCCTCT CCCAACGGCAACCTG


ACATTCGCACCAATG CATCGAACTCCTCTT CCAACGGCAACCTGA


CATTCGCACCAATGC ATCGAACTCCTCTTC CAACGGCAACCTGAG


10ATTCGCACCAATGCT TCGAACTCCTCTTCT AACGGCAACCTGAGT


TTCGCACCAATGCTT CGAACTCCTCTTCTC ACGGCAACCTGAGTT


TCGCACCAATGCTTC GAACTCCTCTTCTCA CGGCAACCTGAGTTA


CGCACCAATGCTTCA AACTCCTCTTCTCAG GGCAACCTGAGTTAC


GCACCAATGCTTCAG ACTCCTCTTCTCAGT GCAACCTGAGTTACT


15CACCAATGCTTCAGT CTCCTCTTCTCAGTT CAACCTGAGTTACTA


ACCAATGCTTCAGTT TCCTCTTCTCAGTTA AACCTGAGTTACTAC


CCAATGCTTCAGTTC CCTCTTCTCAGTTAA ACCTGAGTTACTACA


CAATGCTTCAGTTCC CTCTTCTCAGTTAAT CCTGAGTTACTACAT


AATGCTTCAGTTCCT TCTTCTCAGTTAATC CTGAGTTACTACATT


20ATGCTTCAGTTCCTT CTTCTCAGTTAATCG TGAGTTACTACATTG


TGCTTCAGTTCCTTC TTCTCAGTTAATCGT GAGTTACTACATTGT


GCTTCAGTTCCTTCC TCTCAGTTAATCGTG AGTTACTACATTGTG


CTTCAGTTCCTTCCA CTCAGTTAATCGTGA GTTACTACATTGTGC


TTCAGTTCCTTCCAT TCAGTTAATCGTGAA TTACTACATTGTGCG


25TCAGTTCCTTCCATT CAGTTAATCGTGAAG TACTACATTGTGCGC


CAGTTCCTTCCATTC AGTTAATCGTGAAGT ACTACATTGTGCGCT


AGTTCCTTCCATTCC GTTAATCGTGAAGTG CTACATTGTGCGCTG


GTTCCTTCCATTCCC TTAATCGTGAAGTGG TACATTGTGCGCTGG


TTCCTTCCATTCCCT TAATCGTGAAGTGGA ACATTGTGCGCTGGC


30TCCTTCCATTCCCTT AATCGTGAAGTGGAA CATTGTGCGCTGGCA


CCTTCCATTCCCTTG ATCGTGAAGTGGAAC ATTGTGCGCTGGCAG


CTTCCATTCCCTTGG TCGTGAAGTGGAACC TTGTGCGCTGGCAGC


TTCCATTCCCTTGGA CGTGAAGTGGAACCC TGTGCGCTGGCAGCG


TCCATTCCCTTGGAC GTGAAGTGGAACCCT GTGCGCTGGCAGCGG


35CCATTCCCTTGGACG TGAAGTGGAACCCTC TGCGCTGGCAGCGGC


CATTCCCTTGGACGT GAAGTGGAACCCTCC GCGCTGGCAGCGGCA


ATTCCCTTGGACGTT AAGTGGAACCCTCCC CGCTGGCAGCGGCAG


TTCCCTTGGACGTTC AGTGGAACCCTCCCT GCTGGCAGCGGCAGC


TCCCTTGGACGTTCT GTGGAACCCTCCCTC CTGGCAGCGGCAGCC


40CCCTTGGACGTTCTT TGGAACCCTCCCTCT TGGCAGCGGCAGCCT


CCTTGGACGTTCTTT GGAACCCTCCCTCTC GGCAGCGGCAGCCTC


CTTGGACGTTCTTTC GAACCCTCCCTCTCT GCAGCGGCAGCCTCA


TTGGACGTTCTTTCA AACCCTCCCTCTCTG CAGCGGCAGCCTCAG


TGGACGTTCTTTCAG ACCCTCCCTCTCTGC AGCGGCAGCCTCAGG


45GGACGTTCTTTCAGC CCCTCCCTCTCTGCC GCGGCAGCCTCAGGA


GACGTTCTTTCAGCA CCTCCCTCTCTGCCC CGGCAGCCTCAGGAC


ACGTTCTTTCAGCAT CTCCCTCTCTGCCCA GGCAGCCTCAGGACG


CGTTCTTTCAGCATC TCCCTCTCTGCCCAA GCAGCCTCAGGACGG


GTTCTTTCAGCATCG CCCTCTCTGCCCAAC CAGCCTCAGGACGGC


50TTCTTTCAGCATCGA CCTCTCTGCCCAACG AGCCTCAGGACGGCT


TCTTTCAGCATCGAA CTCTCTGCCCAACGG GCCTCAGGACGGCTA




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CCTCAGGACGGCTAC CCCATCAGGAAGTAT AACCCCAAGACTGAG


CTCAGGACGGCTACC CCATCAGGAAGTATG ACCCCAAGACTGAGG


TCAGGACGGCTACCT CATCAGGAAGTATGC CCCCAAGACTGAGGT


CAGGACGGCTACCTT ATCAGGAAGTATGCC CCCAAGACTGAGGTG


AGGACGGCTACCTTT TCAGGAAGTATGCCG CCAAGACTGAGGTGT


GGACGGCTACCTTTA CAGGAAGTATGCCGA CAAGACTGAGGTGTG


GACGGCTACCTTTAC AGGAAGTATGCCGAC AAGACTGAGGTGTGT


ACGGCTACCTTTACC GGAAGTATGCCGACG AGACTGAGGTGTGTG


CGGCTACCTTTACCG GAAGTATGCCGACGG GACTGAGGTGTGTGG


10GGCTACCTTTACCGG AAGTATGCCGACGGC ACTGAGGTGTGTGGT


GCTACCTTTACCGGC AGTATGCCGACGGCA CTGAGGTGTGTGGTG


CTACCTTTACCGGCA GTATGCCGACGGCAC TGAGGTGTGTGGTGG


TACCTTTACCGGCAC TATGCCGACGGCACC GAGGTGTGTGGTGGG


ACCTTTACCGGCACA ATGCCGACGGCACCA AGGTGTGTGGTGGGG


15CCTTTACCGGCACAA TGCCGACGGCACCAT GGTGTGTGGTGGGGA


CTTTACCGGCACAAT GCCGACGGCACCATC GTGTGTGGTGGGGAG


TTTACCGGCACAATT CCGACGGCACCATCG TGTGTGGTGGGGAGA


TTACCGGCACAATTA CGACGGCACCATCGA GTGTGGTGGGGAGAA


TACCGGCACAATTAC GACGGCACCATCGAC TGTGGTGGGGAGAAA


20ACCGGCACAATTACT ACGGCACCATCGACA GTGGTGGGGAGAAAG


CCGGCACAATTACTG CGGCACCATCGACAT TGGTGGGGAGAAAGG


CGGCACAATTACTGC GGCACCATCGACATT GGTGGGGAGAAAGGG


GGCACAATTACTGCT GCACCATCGACATTG GTGGGGAGAAAGGGC


GCACAATTACTGCTC CACCATCGACATTGA TGGGGAGAAAGGGCC


25CACAATTACTGCTCC ACCATCGACATTGAG GGGGAGAAAGGGCCT


ACAATTACTGCTCCA CCATCGACATTGAGG GGGAGAAAGGGCCTT


CAATTACTGCTCCAA CATCGACATTGAGGA GGAGAAAGGGCCTTG


AATTACTGCTCCAAA ATCGACATTGAGGAG GAGAAAGGGCCTTGC


ATTACTGCTCCAAAG TCGACATTGAGGAGG AGAAAGGGCCTTGCT


30TTACTGCTCCAAAGA CGACATTGAGGAGGT GAAAGGGCCTTGCTG


TACTGCTCCAAAGAC GACATTGAGGAGGTC AAAGGGCCTTGCTGC


ACTGCTCCAAAGACA ACATTGAGGAGGTCA AAGGGCCTTGCTGCG


CTGCTCCAAAGACAA CATTGAGGAGGTCAC AGGGCCTTGCTGCGC


TGCTCCAAAGACAAA ATTGAGGAGGTCACA GGGCCTTGCTGCGCC


35GCTCCAAAGACAAAA TTGAGGAGGTCACAG GGCCTTGCTGCGCCT


CTCCAAAGACAAAAT TGAGGAGGTCACAGA GCCTTGCTGCGCCTG


TCCAAAGACAAAATC GAGGAGGTCACAGAG CCTTGCTGCGCCTGC


CCAAAGACAAAATCC AGGAGGTCACAGAGA CTTGCTGCGCCTGCC


CAAAGACAAAATCCC GGAGGTCACAGAGAA TTGCTGCGCCTGCCC


40AAAGACAAAATCCCC GAGGTCACAGAGAAC TGCTGCGCCTGCCCC


AAGACAAAATCCCCA AGGTCACAGAGAACC GCTGCGCCTGCCCCA


AGACAAAATCCCCAT GGTCACAGAGAACCC CTGCGCCTGCCCCAA


GACAAAATCCCCATC GTCACAGAGAACCCC TGCGCCTGCCCCAAA


ACAAAATCCCCATCA TCACAGAGAACCCCA GCGCCTGCCCCAAAA


45CAAAATCCCCATCAG CACAGAGAACCCCAA CGCCTGCCCCAAAAC


AAAATCCCCATCAGG ACAGAGAACCCCAAG GCCTGCCCCAAAACT


AAATCCCCATCAGGA CAGAGAACCCCAAGA CCTGCCCCAAAACTG


AATCCCCATCAGGAA AGAGAACCCCAAGAC CTGCCCCAAAACTGA


ATCCCCATCAGGAAG GAGAACCCCAAGACT TGCCCCAAAACTGAA


50TCCCCATCAGGAAGT AGAACCCCAAGACTG GCCCCAAAACTGAAG


CCCCATCAGGAAGTA GAACCCCAAGACTGA CCCCAAAACTGAAGC




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CCCAAAACTGAAGCC AAAGTCTTTGAGAAT AGGAAGCGGAGAGAT


CCAAAACTGAAGCCG AAGTCTTTGAGAATT GGAAGCGGAGAGATG


CAAAACTGAAGCCGA AGTCTTTGAGAATTT GAAGCGGAGAGATGT


AAAACTGAAGCCGAG GTCTTTGAGAATTTC AAGCGGAGAGATGTC


AAACTGAAGCCGAGA TCTTTGAGAATTTCC AGCGGAGAGATGTCA


AACTGAAGCCGAGAA CTTTGAGAATTTCCT GCGGAGAGATGTCAT


ACTGAAGCCGAGAAG TTTGAGAATTTCCTG CGGAGAGATGTCATG


CTGAAGCCGAGAAGC TTGAGAATTTCCTGC GGAGAGATGTCATGC


TGAAGCCGAGAAGCA TGAGAATTTCCTGCA GAGAGATGTCATGCA


10GAAGCCGAGAAGCAG GAGAATTTCCTGCAC AGAGATGTCATGCAA


AAGCCGAGAAGCAGG AGAATTTCCTGCACA GAGATGTCATGCAAG


AGCCGAGAAGCAGGC GAATTTCCTGCACAA AGATGTCATGCAAGT


GCCGAGAAGCAGGCC AATTTCCTGCACAAC GATGTCATGCAAGTG


CCGAGAAGCAGGCCG ATTTCCTGCACAACT ATGTCATGCAAGTGG


15CGAGAAGCAGGCCGA TTTCCTGCACAACTC TGTCATGCAAGTGGC


GAGAAGCAGGCCGAG TTCCTGCACAACTCC GTCATGCAAGTGGCC


AGAAGCAGGCCGAGA TCCTGCACAACTCCA TCATGCAAGTGGCCA


GAAGCAGGCCGAGAA CCTGCACAACTCCAT CATGCAAGTGGCCAA


AAGCAGGCCGAGAAG CTGCACAACTCCATC ATGCAAGTGGCCAAC


20AGCAGGCCGAGAAGG TGCACAACTCCATCT TGCAAGTGGCCAACA


GCAGGCCGAGAAGGA GCACAACTCCATCTT GCAAGTGGCCAACAC


CAGGCCGAGAAGGAG CACAACTCCATCTTC CAAGTGGCCAACACC


AGGCCGAGAAGGAGG ACAACTCCATCTTCG AAGTGGCCAACACCA


GGCCGAGAAGGAGGA CAACTCCATCTTCGT AGTGGCCAACACCAC


25GCCGAGAAGGAGGAG AACTCCATCTTCGTG GTGGCCAACACCACC


CCGAGAAGGAGGAGG ACTCCATCTTCGTGC TGGCCAACACCACCA


CGAGAAGGAGGAGGC CTCCATCTTCGTGCC GGCCAACACCACCAT


GAGAAGGAGGAGGCT TCCATCTTCGTGCCC GCCAACACCACCATG


AGAAGGAGGAGGCTG CCATCTTCGTGCCCA CCAACACCACCATGT


30GAAGGAGGAGGCTGA CATCTTCGTGCCCAG CAACACCACCATGTC


AAGGAGGAGGCTGAA ATCTTCGTGCCCAGA AACACCACCATGTCC


AGGAGGAGGCTGAAT TCTTCGTGCCCAGAC ACACCACCATGTCCA


GGAGGAGGCTGAATA CTTCGTGCCCAGACC CACCACCATGTCCAG


GAGGAGGCTGAATAC TTCGTGCCCAGACCT ACCACCATGTCCAGC


35AGGAGGCTGAATACC TCGTGCCCAGACCTG CCACCATGTCCAGCC


GGAGGCTGAATACCG CGTGCCCAGACCTGA CACCATGTCCAGCCG


GAGGCTGAATACCGC GTGCCCAGACCTGAA ACCATGTCCAGCCGA


AGGCTGAATACCGCA TGCCCAGACCTGAAA CCATGTCCAGCCGAA


GGCTGAATACCGCAA GCCCAGACCTGAAAG CATGTCCAGCCGAAG


40GCTGAATACCGCAAA CCCAGACCTGAAAGG ATGTCCAGCCGAAGC


CTGAATACCGCAAAG CCAGACCTGAAAGGA TGTCCAGCCGAAGCA


TGAATACCGCAAAGT CAGACCTGAAAGGAA GTCCAGCCGAAGCAG


GAATACCGCAAAGTC AGACCTGAAAGGAAG TCCAGCCGAAGCAGG


AATACCGCAAAGTCT GACCTGAAAGGAAGC CCAGCCGAAGCAGGA


45ATACCGCAAAGTCTT ACCTGAAAGGAAGCG CAGCCGAAGCAGGAA


TACCGCAAAGTCTTT CCTGAAAGGAAGCGG AGCCGAAGCAGGAAC


ACCGCAAAGTCTTTG CTGAAAGGAAGCGGA GCCGAAGCAGGAACA


CCGCAAAGTCTTTGA TGAAAGGAAGCGGAG CCGAAGCAGGAACAC


CGCAAAGTCTTTGAG GAAAGGAAGCGGAGA CGAAGCAGGAACACC


50GCAAAGTCTTTGAGA AAAGGAAGCGGAGAG GAAGCAGGAACACCA


CAAAGTCTTTGAGAA AAGGAAGCGGAGAGA AAGCAGGAACACCAC




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AGCAGGAACACCACG CTGGAGACAGAGTAC ACTGTCATTTCTAAC


GCAGGAACACCACGG TGGAGACAGAGTACC CTGTCATTTCTAACC


CAGGAACACCACGGC GGAGACAGAGTACCC TGTCATTTCTAACCT


AGGAACACCACGGCC GAGACAGAGTACCCT GTCATTTCTAACCTT


GGAACACCACGGCCG AGACAGAGTACCCTT TCATTTCTAACCTTC


GAACACCACGGCCGC GACAGAGTACCCTTT CATTTCTAACCTTCG


AACACCACGGCCGCA ACAGAGTACCCTTTC ATTTCTAACCTTCGG


ACACCACGGCCGCAG CAGAGTACCCTTTCT TTTCTAACCTTCGGC


CACCACGGCCGCAGA AGAGTACCCTTTCTT TTCTAACCTTCGGCC


10ACCACGGCCGCAGAC GAGTACCCTTTCTTT TCTAACCTTCGGCCT


CCACGGCCGCAGACA AGTACCCTTTCTTTG CTAACCTTCGGCCTT


CACGGCCGCAGACAC GTACCCTTTCTTTGA TAACCTTCGGCCTTT


ACGGCCGCAGACACC TACCCTTTCTTTGAG AACCTTCGGCCTTTC


CGGCCGCAGACACCT ACCCTTTCTTTGAGA ACCTTCGGCCTTTCA


15GGCCGCAGACACCTA CCCTTTCTTTGAGAG CCTTCGGCCTTTCAC


GCCGCAGACACCTAC CCTTTCTTTGAGAGC CTTCGGCCTTTCACA


CCGCAGACACCTACA CTTTCTTTGAGAGCA TTCGGCCTTTCACAT


CGCAGACACCTACAA TTTCTTTGAGAGCAG TCGGCCTTTCACATT


GCAGACACCTACAAC TTCTTTGAGAGCAGA CGGCCTTTCACATTG


20CAGACACCTACAACA TCTTTGAGAGCAGAG GGCCTTTCACATTGT


AGACACCTACAACAT CTTTGAGAGCAGAGT GCCTTTCACATTGTA


GACACCTACAACATC TTTGAGAGCAGAGTG CCTTTCACATTGTAC


ACACCTACAACATCA TTGAGAGCAGAGTGG CTTTCACATTGTACC


CACCTACAACATCAC TGAGAGCAGAGTGGA TTTCACATTGTACCG


25ACCTACAACATCACC GAGAGCAGAGTGGAT TTCACATTGTACCGC


CCTACAACATCACCG AGAGCAGAGTGGATA TCACATTGTACCGCA


CTACAACATCACCGA GAGCAGAGTGGATAA CACATTGTACCGCAT


TACAACATCACCGAC AGCAGAGTGGATAAC ACATTGTACCGCATC


ACAACATCACCGACC GCAGAGTGGATAACA CATTGTACCGCATCG


30CAACATCACCGACCC CAGAGTGGATAACAA ATTGTACCGCATCGA


AACATCACCGACCCG AGAGTGGATAACAAG TTGTACCGCATCGAT


ACATCACCGACCCGG GAGTGGATAACAAGG TGTACCGCATCGATA


CATCACCGACCCGGA AGTGGATAACAAGGA GTACCGCATCGATAT


ATCACCGACCCGGAA GTGGATAACAAGGAG TACCGCATCGATATC


35TCACCGACCCGGAAG TGGATAACAAGGAGA ACCGCATCGATATCC


CACCGACCCGGAAGA GGATAACAAGGAGAG CCGCATCGATATCCA


ACCGACCCGGAAGAG GATAACAAGGAGAGA CGCATCGATATCCAC


CCGACCCGGAAGAGC ATAACAAGGAGAGAA GCATCGATATCCACA


CGACCCGGAAGAGCT TAACAAGGAGAGAAC CATCGATATCCACAG


40GACCCGGAAGAGCTG AACAAGGAGAGAACT ATCGATATCCACAGC


ACCCGGAAGAGCTGG ACAAGGAGAGAACTG TCGATATCCACAGCT


CCCGGAAGAGCTGGA CAAGGAGAGAACTGT CGATATCCACAGCTG


CCGGAAGAGCTGGAG AAGGAGAGAACTGTC GATATCCACAGCTGC


CGGAAGAGCTGGAGA AGGAGAGAACTGTCA ATATCCACAGCTGCA


45GGAAGAGCTGGAGAC GGAGAGAACTGTCAT TATCCACAGCTGCAA


GAAGAGCTGGAGACA GAGAGAACTGTCATT ATCCACAGCTGCAAC


AAGAGCTGGAGACAG AGAGAACTGTCATTT TCCACAGCTGCAACC


AGAGCTGGAGACAGA GAGAACTGTCATTTC CCACAGCTGCAACCA


GAGCTGGAGACAGAG AGAACTGTCATTTCT CACAGCTGCAACCAC


50AGCTGGAGACAGAGT GAACTGTCATTTCTA ACAGCTGCAACCACG


GCTGGAGACAGAGTA AACTGTCATTTCTAA CAGCTGCAACCACGA




CA 02376284 2001-12-05
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_77_
AGCTGCAACCACGAG TTTGCAAGGACTATG ACCTGGGAGCCAAGG


GCTGCAACCACGAGG TTGCAAGGACTATGC CCTGGGAGCCAAGGC


CTGCAACCACGAGGC TGCAAGGACTATGCC CTGGGAGCCAAGGCC


TGCAACCACGAGGCT GCAAGGACTATGCCC TGGGAGCCAAGGCCT


GCAACCACGAGGCTG CAAGGACTATGCCCG GGGAGCCAAGGCCTG


CAACCACGAGGCTGA AAGGACTATGCCCGC GGAGCCAAGGCCTGA


AACCACGAGGCTGAG AGGACTATGCCCGCA GAGCCAAGGCCTGAA


ACCACGAGGCTGAGA GGACTATGCCCGCAG AGCCAAGGCCTGAAA


CCACGAGGCTGAGAA GACTATGCCCGCAGA GCCAAGGCCTGAAAA


10CACGAGGCTGAGAAG ACTATGCCCGCAGAA CCAAGGCCTGAAAAC


ACGAGGCTGAGAAGC CTATGCCCGCAGAAG CAAGGCCTGAAAACT


CGAGGCTGAGAAGCT TATGCCCGCAGAAGG AAGGCCTGAAAACTC


GAGGCTGAGAAGCTG ATGCCCGCAGAAGGA AGGCCTGAAAACTCC


AGGCTGAGAAGCTGG TGCCCGCAGAAGGAG GGCCTGAAAACTCCA


15GGCTGAGAAGCTGGG GCCCGCAGAAGGAGC GCCTGAAAACTCCAT


GCTGAGAAGCTGGGC CCCGCAGAAGGAGCA CCTGAAAACTCCATC


CTGAGAAGCTGGGCT CCGCAGAAGGAGCAG CTGAAAACTCCATCT


TGAGAAGCTGGGCTG CGCAGAAGGAGCAGA TGAAAACTCCATCTT


GAGAAGCTGGGCTGC GCAGAAGGAGCAGAT GAAAACTCCATCTTT


20AGAAGCTGGGCTGCA CAGAAGGAGCAGATG AAAACTCCATCTTTT


GAAGCTGGGCTGCAG AGAAGGAGCAGATGA AAACTCCATCTTTTT


AAGCTGGGCTGCAGC GAAGGAGCAGATGAC AACTCCATCTTTTTA


AGCTGGGCTGCAGCG AAGGAGCAGATGACA ACTCCATCTTTTTAA


GCTGGGCTGCAGCGC AGGAGCAGATGACAT CTCCATCTTTTTAAA


25CTGGGCTGCAGCGCC GGAGCAGATGACATT TCCATCTTTTTAAAG


TGGGCTGCAGCGCCT GAGCAGATGACATTC CCATCTTTTTAAAGT


GGGCTGCAGCGCCTC AGCAGATGACATTCC CATCTTTTTAAAGTG


GGCTGCAGCGCCTCC GCAGATGACATTCCT ATCTTTTTAAAGTGG


GCTGCAGCGCCTCCA CAGATGACATTCCTG TCTTTTTAAAGTGGC


30CTGCAGCGCCTCCAA AGATGACATTCCTGG CTTTTTAAAGTGGCC


TGCAGCGCCTCCAAC GATGACATTCCTGGG TTTTTAAAGTGGCCG


GCAGCGCCTCCAACT ATGACATTCCTGGGC TTTTAAAGTGGCCGG


CAGCGCCTCCAACTT TGACATTCCTGGGCC TTTAAAGTGGCCGGA


AGCGCCTCCAACTTC GACATTCCTGGGCCA TTAAAGTGGCCGGA.A


35GCGCCTCCAACTTCG ACATTCCTGGGCCAG TAAAGTGGCCGGAAC


CGCCTCCAACTTCGT CATTCCTGGGCCAGT AAAGTGGCCGGAACC


GCCTCCAACTTCGTC ATTCCTGGGCCAGTG AAGTGGCCGGAACCT


CCTCCAACTTCGTCT TTCCTGGGCCAGTGA AGTGGCCGGAACCTG


CTCCAACTTCGTCTT TCCTGGGCCAGTGAC GTGGCCGGAACCTGA


40TCCAACTTCGTCTTT CCTGGGCCAGTGACC TGGCCGGAACCTGAG


CCAACTTCGTCTTTG CTGGGCCAGTGACCT GGCCGGAACCTGAGA


CAACTTCGTCTTTGC TGGGCCAGTGACCTG GCCGGAACCTGAGAA


AACTTCGTCTTTGCA GGGCCAGTGACCTGG CCGGAACCTGAGAAT


ACTTCGTCTTTGCAA GGCCAGTGACCTGGG CGGAACCTGAGAATC


45CTTCGTCTTTGCAAG GCCAGTGACCTGGGA GGAACCTGAGAATCC


TTCGTCTTTGCAAGG CCAGTGACCTGGGAG GAACCTGAGAATCCC


TCGTCTTTGCAAGGA CAGTGACCTGGGAGC AACCTGAGAATCCCA


CGTCTTTGCAAGGAC AGTGACCTGGGAGCC ACCTGAGAATCCCAA


GTCTTTGCAAGGACT GTGACCTGGGAGCCA CCTGAGAATCCCAAT


50TCTTTGCAAGGACTA TGACCTGGGAGCCAA CTGAGAATCCCAATG


CTTTGCAAGGACTAT GACCTGGGAGCCAAG TGAGAATCCCAATGG




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GAGAATCCCAATGGA GTTGAGGATCAGCGA GGGGCCAAGCTAAAC


AGAATCCCAATGGAT TTGAGGATCAGCGAG GGGCCAAGCTAAACC


GAATCCCAATGGATT TGAGGATCAGCGAGA GGCCAAGCTAA.P.CCG


AATCCCAATGGATTG GAGGATCAGCGAGAA GCCAAGCTAAACCGG


ATCCCAATGGATTGA AGGATCAGCGAGAAT CCAAGCTAAACCGGC


TCCCAATGGATTGAT GGATCAGCGAGAATG CAAGCTAAACCGGCT


CCCAATGGATTGATT GATCAGCGAGAATGT AAGCTAAACCGGCTA


CCAATGGATTGATTC ATCAGCGAGAATGTG AGCTAAACCGGCTAA


CAATGGATTGATTCT TCAGCGAGAATGTGT GCTAAACCGGCTAAA


10AATGGATTGATTCTA CAGCGAGAATGTGTG CTAAACCGGCTAAAC


ATGGATTGATTCTAA AGCGAGAATGTGTGT TAAACCGGCTAAACC


TGGATTGATTCTAAT GCGAGAATGTGTGTC AAACCGGCTAAACCC


GGATTGATTCTAATG CGAGAATGTGTGTCC AACCGGCTAAACCCG


GATTGATTCTAATGT GAGAATGTGTGTCCA ACCGGCTAAACCCGG


15ATTGATTCTAATGTA AGAATGTGTGTCCAG CCGGCTAAACCCGGG


TTGATTCTAATGTAT GAATGTGTGTCCAGA CGGCTAA.ACCCGGGG


TGATTCTAATGTATG AATGTGTGTCCAGAC GGCTAAACCCGGGGA


GATTCTAATGTATGA ATGTGTGTCCAGACA GCTAAACCCGGGGAA


ATTCTAATGTATGAA TGTGTGTCCAGACAG CTAAACCCGGGGAAC


20TTCTAATGTATGAAA GTGTGTCCAGACAGG TAAACCCGGGGAACT


TCTAATGTATGAAAT TGTGTCCAGACAGGA AAACCCGGGGAACTA


CTAATGTATGAAATA GTGTCCAGACAGGAA AACCCGGGGAACTAC


TAATGTATGAAATAA TGTCCAGACAGGAAT ACCCGGGGAACTACA


AATGTATGAAATAAA GTCCAGACAGGAATA CCCGGGGAACTACAC


25ATGTATGAAATAAAA TCCAGACAGGAATAC CCGGGGAACTACACA


TGTATGAAATAAAAT CCAGACAGGAATACA CGGGGAACTACACAG


GTATGAAATAAAATA CAGACAGGAATACAG GGGGAACTACACAGC


TATGAA.ATAAAATAC AGACAGGAATACAGG GGGAACTACACAGCC


ATGAAATAAAATACG GACAGGAATACAGGA GGAACTACACAGCCC


30TGAAATAAAATACGG ACAGGAATACAGGAA GAACTACACAGCCCG


GAAATAAAATACGGA CAGGAATACAGGAAG AACTACACAGCCCGG


AAATAAAATACGGAT AGGAATACAGGAAGT ACTACACAGCCCGGA


AATAAAATACGGATC GGAATACAGGAAGTA CTACACAGCCCGGAT


ATAAAATACGGATCA GAATACAGGAAGTAT TACACAGCCCGGATT


35TAAAATACGGATCAC AATACAGGAAGTATG ACACAGCCCGGATTC


AAAATACGGATCACA ATACAGGAAGTATGG CACAGCCCGGATTCA


AAATACGGATCACAA TACAGGAAGTATGGA ACAGCCCGGATTCAG


AATACGGATCACAAG ACAGGAAGTATGGAG CAGCCCGGATTCAGG


ATACGGATCACAAGT CAGGAAGTATGGAGG AGCCCGGATTCAGGC


40TACGGATCACAAGTT AGGAAGTATGGAGGG GCCCGGATTCAGGCC


ACGGATCACAAGTTG GGAAGTATGGAGGGG CCCGGATTCAGGCCA


CGGATCACAAGTTGA GAAGTATGGAGGGGC CCGGATTCAGGCCAC


GGATCACAAGTTGAG AAGTATGGAGGGGCC CGGATTCAGGCCACA


GATCACAAGTTGAGG AGTATGGAGGGGCCA GGATTCAGGCCACAT


45ATCACAAGTTGAGGA GTATGGAGGGGCCAA GATTCAGGCCACATC


TCACAAGTTGAGGAT TATGGAGGGGCCAAG ATTCAGGCCACATCT


CACAAGTTGAGGATC ATGGAGGGGCCAAGC TTCAGGCCACATCTC


ACAAGTTGAGGATCA TGGAGGGGCCAAGCT TCAGGCCACATCTCT


CAAGTTGAGGATCAG GGAGGGGCCAAGCTA CAGGCCACATCTCTC


50AAGTTGAGGATCAGC GAGGGGCCAAGCTAA AGGCCACATCTCTCT


AGTTGAGGATCAGCG AGGGGCCAAGCTAAA GGCCACATCTCTCTC




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GCCACATCTCTCTCT GTCCAGGCCAAAACA CCCGTCGCTGTCCTG


CCACATCTCTCTCTG TCCAGGCCAAAACAG CCGTCGCTGTCCTGT


CACATCTCTCTCTGG CCAGGCCAAAACAGG CGTCGCTGTCCTGTT


ACATCTCTCTCTGGG CAGGCCAAAACAGGA GTCGCTGTCCTGTTG


CATCTCTCTCTGGGA AGGCCAAAACAGGAT TCGCTGTCCTGTTGA


ATCTCTCTCTGGGAA GGCCAAAACAGGATA CGCTGTCCTGTTGAT


TCTCTCTCTGGGAAT GCCAAAACAGGATAT GCTGTCCTGTTGATC


CTCTCTCTGGGAATG CCAAAACAGGATATG CTGTCCTGTTGATCG


TCTCTCTGGGAATGG CAAAACAGGATATGA TGTCCTGTTGATCGT


10CTCTCTGGGAATGGG AAAACAGGATATGAA GTCCTGTTGATCGTG


TCTCTGGGAATGGGT AAACAGGATATGAAA TCCTGTTGATCGTGG


CTCTGGGAATGGGTC AACAGGATATGAAAA CCTGTTGATCGTGGG


TCTGGGAATGGGTCG ACAGGATATGAAAAC CTGTTGATCGTGGGA


CTGGGAATGGGTCGT CAGGATATGAAAACT TGTTGATCGTGGGAG


15TGGGAATGGGTCGTG AGGATATGAAAACTT GTTGATCGTGGGAGG


GGGAATGGGTCGTGG GGATATGAAAACTTC TTGATCGTGGGAGGG


GGAATGGGTCGTGGA GATATGAAAACTTCA TGATCGTGGGAGGGT


GAATGGGTCGTGGAC ATATGAAAACTTCAT GATCGTGGGAGGGTT


AATGGGTCGTGGACA TATGAAAACTTCATC ATCGTGGGAGGGTTG


20ATGGGTCGTGGACAG ATGAAAACTTCATCC TCGTGGGAGGGTTGG


TGGGTCGTGGACAGA TGAAAACTTCATCCA CGTGGGAGGGTTGGT


GGGTCGTGGACAGAT GAAAACTTCATCCAT GTGGGAGGGTTGGTG


GGTCGTGGACAGATC AAAACTTCATCCATC TGGGAGGGTTGGTGA


GTCGTGGACAGATCC AAACTTCATCCATCT GGGAGGGTTGGTGAT


25TCGTGGACAGATCCT AACTTCATCCATCTG GGAGGGTTGGTGATT


CGTGGACAGATCCTG ACTTCATCCATCTGA GAGGGTTGGTGATTA


GTGGACAGATCCTGT CTTCATCCATCTGAT AGGGTTGGTGATTAT


TGGACAGATCCTGTG TTCATCCATCTGATC GGGTTGGTGATTATG


GGACAGATCCTGTGT TCATCCATCTGATCA GGTTGGTGATTATGC


30GACAGATCCTGTGTT CATCCATCTGATCAT GTTGGTGATTATGCT


ACAGATCCTGTGTTC ATCCATCTGATCATC TTGGTGATTATGCTG


CAGATCCTGTGTTCT TCCATCTGATCATCG TGGTGATTATGCTGT


AGATCCTGTGTTCTT CCATCTGATCATCGC GGTGATTATGCTGTA


GATCCTGTGTTCTTC CATCTGATCATCGCT GTGATTATGCTGTAC


35ATCCTGTGTTCTTCT ATCTGATCATCGCTC TGATTATGCTGTACG


TCCTGTGTTCTTCTA TCTGATCATCGCTCT GATTATGCTGTACGT


CCTGTGTTCTTCTAT CTGATCATCGCTCTG ATTATGCTGTACGTC


CTGTGTTCTTCTATG TGATCATCGCTCTGC TTATGCTGTACGTCT


TGTGTTCTTCTATGT GATCATCGCTCTGCC TATGCTGTACGTCTT


40GTGTTCTTCTATGTC ATCATCGCTCTGCCC ATGCTGTACGTCTTC


TGTTCTTCTATGTCC TCATCGCTCTGCCCG TGCTGTACGTCTTCC


GTTCTTCTATGTCCA CATCGCTCTGCCCGT GCTGTACGTCTTCCA


TTCTTCTATGTCCAG ATCGCTCTGCCCGTC CTGTACGTCTTCCAT


TCTTCTATGTCCAGG TCGCTCTGCCCGTCG TGTACGTCTTCCATA


45CTTCTATGTCCAGGC CGCTCTGCCCGTCGC GTACGTCTTCCATAG


TTCTATGTCCAGGCC GCTCTGCCCGTCGCT TACGTCTTCCATAGA


TCTATGTCCAGGCCA CTCTGCCCGTCGCTG ACGTCTTCCATAGAA


CTATGTCCAGGCCAA TCTGCCCGTCGCTGT CGTCTTCCATAGAAA


TATGTCCAGGCCAAA CTGCCCGTCGCTGTC GTCTTCCATAGAAAG


50ATGTCCAGGCCAAAA TGCCCGTCGCTGTCC TCTTCCATAGAAAGA


TGTCCAGGCCAAAAC GCCCGTCGCTGTCCT CTTCCATAGAAAGAG




CA 02376284 2001-12-05
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-80-
TTCCATAGAAAGAGA TCTGTGAACCCGGAG TGGGAGGTGGCTCGG


TCCATAGAAAGAGAA CTGTGAACCCGGAGT GGGAGGTGGCTCGGG


CCATAGAAAGAGAAA TGTGAACCCGGAGTA GGAGGTGGCTCGGGA


CATAGAAAGAGAAAT GTGAACCCGGAGTAC GAGGTGGCTCGGGAG


ATAGAAAGAGAAATA TGAACCCGGAGTACT AGGTGGCTCGGGAGA


TAGAAAGAGAAATAA GAACCCGGAGTACTT GGTGGCTCGGGAGAA


AGAAAGAGAAATAAC AACCCGGAGTACTTC GTGGCTCGGGAGAAG


GAAAGAGAAATAACA ACCCGGAGTACTTCA TGGCTCGGGAGAAGA


AAAGAGAAATAACAG CCCGGAGTACTTCAG GGCTCGGGAGAAGAT


10AAGAGAAATAACAGC CCGGAGTACTTCAGC GCTCGGGAGAAGATC


AGAGAAATAACAGCA CGGAGTACTTCAGCG CTCGGGAGAAGATCA


GAGAAATAACAGCAG GGAGTACTTCAGCGC TCGGGAGAAGATCAC


AGAAATAACAGCAGG GAGTACTTCAGCGCT CGGGAGAAGATCACC


GAAATAACAGCAGGC AGTACTTCAGCGCTG GGGAGAAGATCACCA


15AAATAACAGCAGGCT GTACTTCAGCGCTGC GGAGAAGATCACCAT


AATAACAGCAGGCTG TACTTCAGCGCTGCT GAGAAGATCACCATG


ATAACAGCAGGCTGG ACTTCAGCGCTGCTG AGAAGATCACCATGA


TAACAGCAGGCTGGG CTTCAGCGCTGCTGA GAAGATCACCATGAG


AACAGCAGGCTGGGG TTCAGCGCTGCTGAT AAGATCACCATGAGC


20ACAGCAGGCTGGGGA TCAGCGCTGCTGATG AGATCACCATGAGCC


CAGCAGGCTGGGGAA CAGCGCTGCTGATGT GATCACCATGAGCCG


AGCAGGCTGGGGAAT AGCGCTGCTGATGTG ATCACCATGAGCCGG


GCAGGCTGGGGAATG GCGCTGCTGATGTGT TCACCATGAGCCGGG


CAGGCTGGGGAATGG CGCTGCTGATGTGTA CACCATGAGCCGGGA


25AGGCTGGGGAATGGA GCTGCTGATGTGTAC ACCATGAGCCGGGAA


GGCTGGGGAATGGAG CTGCTGATGTGTACG CCATGAGCCGGGAAC


GCTGGGGAATGGAGT TGCTGATGTGTACGT CATGAGCCGGGAACT


CTGGGGAATGGAGTG GCTGATGTGTACGTT ATGAGCCGGGAACTT


TGGGGAATGGAGTGC CTGATGTGTACGTTC TGAGCCGGGAACTTG


30GGGGAATGGAGTGCT TGATGTGTACGTTCC GAGCCGGGAACTTGG


GGGAATGGAGTGCTG GATGTGTACGTTCCT AGCCGGGAACTTGGG


GGAATGGAGTGCTGT ATGTGTACGTTCCTG GCCGGGAACTTGGGC


GAATGGAGTGCTGTA TGTGTACGTTCCTGA CCGGGAACTTGGGCA


AATGGAGTGCTGTAT GTGTACGTTCCTGAT CGGGAACTTGGGCAG


35ATGGAGTGCTGTATG TGTACGTTCCTGATG GGGAACTTGGGCAGG


TGGAGTGCTGTATGC GTACGTTCCTGATGA GGAACTTGGGCAGGG


GGAGTGCTGTATGCC TACGTTCCTGATGAG GAACTTGGGCAGGGG


GAGTGCTGTATGCCT ACGTTCCTGATGAGT AACTTGGGCAGGGGT


AGTGCTGTATGCCTC CGTTCCTGATGAGTG ACTTGGGCAGGGGTC


40GTGCTGTATGCCTCT GTTCCTGATGAGTGG CTTGGGCAGGGGTCG


TGCTGTATGCCTCTG TTCCTGATGAGTGGG TTGGGCAGGGGTCGT


GCTGTATGCCTCTGT TCCTGATGAGTGGGA TGGGCAGGGGTCGTT


CTGTATGCCTCTGTG CCTGATGAGTGGGAG GGGCAGGGGTCGTTT


TGTATGCCTCTGTGA CTGATGAGTGGGAGG GGCAGGGGTCGTTTG


45GTATGCCTCTGTGAA TGATGAGTGGGAGGT GCAGGGGTCGTTTGG


TATGCCTCTGTGAAC GATGAGTGGGAGGTG CAGGGGTCGTTTGGG


ATGCCTCTGTGAACC ATGAGTGGGAGGTGG AGGGGTCGTTTGGGA


TGCCTCTGTGAACCC TGAGTGGGAGGTGGC GGGGTCGTTTGGGAT


GCCTCTGTGAACCCG GAGTGGGAGGTGGCT GGGTCGTTTGGGATG


SOCCTCTGTGAACCCGG AGTGGGAGGTGGCTC GGTCGTTTGGGATGG


CTCTGTGAACCCGGA GTGGGAGGTGGCTCG GTCGTTTGGGATGGT




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TCGTTTGGGATGGTC CCTGAAACCAGAGTG GAGAGGATTGAGTTT


CGTTTGGGATGGTCT CTGAAACCAGAGTGG AGAGGATTGAGTTTC


GTTTGGGATGGTCTA TGAAACCAGAGTGGC GAGGATTGAGTTTCT


TTTGGGATGGTCTAT GAAACCAGAGTGGCC AGGATTGAGTTTCTC


TTGGGATGGTCTATG AAACCAGAGTGGCCA GGATTGAGTTTCTCA


TGGGATGGTCTATGA AACCAGAGTGGCCAT GATTGAGTTTCTCAA


GGGATGGTCTATGAA ACCAGAGTGGCCATT ATTGAGTTTCTCAAC


GGATGGTCTATGAAG CCAGAGTGGCCATTA TTGAGTTTCTCAACG


GATGGTCTATGAAGG CAGAGTGGCCATTAA TGAGTTTCTCAACGA


10ATGGTCTATGAAGGA AGAGTGGCCATTAAA GAGTTTCTCAACGAA


TGGTCTATGAAGGAG GAGTGGCCATTAAAA AGTTTCTCAACGAAG


GGTCTATGAAGGAGT AGTGGCCATTAAAAC GTTTCTCAACGAAGC


GTCTATGAAGGAGTT GTGGCCATTAAAACA TTTCTCAACGAAGCT


TCTATGAAGGAGTTG TGGCCATTAAAACAG TTCTCAACGAAGCTT


15CTATGAAGGAGTTGC GGCCATTAAAACAGT TCTCAACGAAGCTTC


TATGAAGGAGTTGCC GCCATTAAAACAGTG CTCAACGAAGCTTCT


ATGAAGGAGTTGCCA CCATTAAAACAGTGA TCAACGAAGCTTCTG


TGAAGGAGTTGCCAA CATTAAAACAGTGAA CAACGAAGCTTCTGT


GAAGGAGTTGCCAAG ATTAAAACAGTGAAC AACGAAGCTTCTGTG


20AAGGAGTTGCCAAGG TTAAAACAGTGAACG ACGAAGCTTCTGTGA


AGGAGTTGCCAAGGG TAAAACAGTGAACGA CGAAGCTTCTGTGAT


GGAGTTGCCAAGGGT AAAACAGTGAACGAG GAAGCTTCTGTGATG


GAGTTGCCAAGGGTG AAACAGTGAACGAGG AAGCTTCTGTGATGA


AGTTGCCAAGGGTGT AACAGTGAACGAGGC AGCTTCTGTGATGAA


25GTTGCCAAGGGTGTG ACAGTGAACGAGGCC GCTTCTGTGATGAAG


TTGCCAAGGGTGTGG CAGTGAACGAGGCCG CTTCTGTGATGAAGG


TGCCAAGGGTGTGGT AGTGAACGAGGCCGC TTCTGTGATGAAGGA


GCCAAGGGTGTGGTG GTGAACGAGGCCGCA TCTGTGATGAAGGAG


CCAAGGGTGTGGTGA TGAACGAGGCCGCAA CTGTGATGAAGGAGT


30CAAGGGTGTGGTGAA GAACGAGGCCGCAAG TGTGATGAAGGAGTT


AAGGGTGTGGTGAAA AACGAGGCCGCAAGC GTGATGAAGGAGTTC


AGGGTGTGGTGAAAG ACGAGGCCGCAAGCA TGATGAAGGAGTTCA


GGGTGTGGTGAAAGA CGAGGCCGCAAGCAT GATGAAGGAGTTCAA


GGTGTGGTGAAAGAT GAGGCCGCAAGCATG ATGAAGGAGTTCAAT


35GTGTGGTGAAAGATG AGGCCGCAAGCATGC TGAAGGAGTTCAATT


TGTGGTGAAAGATGA GGCCGCAAGCATGCG GAAGGAGTTCAATTG


GTGGTGAAAGATGAA GCCGCAAGCATGCGT AAGGAGTTCAATTGT


TGGTGAAAGATGAAC CCGCAAGCATGCGTG AGGAGTTCAATTGTC


GGTGAAAGATGAACC CGCAAGCATGCGTGA GGAGTTCAATTGTCA


40GTGAAAGATGAACCT GCAAGCATGCGTGAG GAGTTCAATTGTCAC


TGAAAGATGAACCTG CAAGCATGCGTGAGA AGTTCAATTGTCACC


GAAAGATGAACCTGA AAGCATGCGTGAGAG GTTCAATTGTCACCA


AAAGATGAACCTGAA AGCATGCGTGAGAGG TTCAATTGTCACCAT


AAGATGAACCTGAAA GCATGCGTGAGAGGA TCAATTGTCACCATG


45AGATGAACCTGAAAC CATGCGTGAGAGGAT CAATTGTCACCATGT


GATGAACCTGAAACC ATGCGTGAGAGGATT AATTGTCACCATGTG


ATGAACCTGAAACCA TGCGTGAGAGGATTG ATTGTCACCATGTGG


TGAACCTGAAACCAG GCGTGAGAGGATTGA TTGTCACCATGTGGT


GAACCTGAAACCAGA CGTGAGAGGATTGAG TGTCACCATGTGGTG


50AACCTGAAACCAGAG GTGAGAGGATTGAGT GTCACCATGTGGTGC


ACCTGAAACCAGAGT TGAGAGGATTGAGTT TCACCATGTGGTGCG




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CACCATGTGGTGCGA GTCATCATGGAACTG CTGAGGCCAGAAATG


ACCATGTGGTGCGAT TCATCATGGAACTGA TGAGGCCAGAAATGG


CCATGTGGTGCGATT CATCATGGAACTGAT GAGGCCAGAAATGGA


CATGTGGTGCGATTG ATCATGGAACTGATG AGGCCAGAAATGGAG


ATGTGGTGCGATTGC TCATGGAACTGATGA GGCCAGAAATGGAGA


TGTGGTGCGATTGCT CATGGAACTGATGAC GCCAGAAATGGAGAA


GTGGTGCGATTGCTG ATGGAACTGATGACA CCAGAAATGGAGAAT


TGGTGCGATTGCTGG TGGAACTGATGACAC CAGAAATGGAGAATA


GGTGCGATTGCTGGG GGAACTGATGACACG AGAAATGGAGAATAA


10GTGCGATTGCTGGGT GAACTGATGACACGG GAAATGGAGAATAAT


TGCGATTGCTGGGTG AACTGATGACACGGG AAATGGAGAATAATC


GCGATTGCTGGGTGT ACTGATGACACGGGG AATGGAGAATAATCC


CGATTGCTGGGTGTG CTGATGACACGGGGC ATGGAGAATAATCCA


GATTGCTGGGTGTGG TGATGACACGGGGCG TGGAGAATAATCCAG


15ATTGCTGGGTGTGGT GATGACACGGGGCGA GGAGAATAATCCAGT


TTGCTGGGTGTGGTG ATGACACGGGGCGAT GAGAATAATCCAGTC


TGCTGGGTGTGGTGT TGACACGGGGCGATC AGAATAATCCAGTCC


GCTGGGTGTGGTGTC GACACGGGGCGATCT GAATAATCCAGTCCT


CTGGGTGTGGTGTCC ACACGGGGCGATCTC AATAATCCAGTCCTA


20TGGGTGTGGTGTCCC CACGGGGCGATCTCA ATAATCCAGTCCTAG


GGGTGTGGTGTCCCA ACGGGGCGATCTCAA TAATCCAGTCCTAGC


GGTGTGGTGTCCCAA CGGGGCGATCTCAAA AATCCAGTCCTAGCA


GTGTGGTGTCCCAAG GGGGCGATCTCAAAA ATCCAGTCCTAGCAC


TGTGGTGTCCCAAGG GGGCGATCTCAAAAG TCCAGTCCTAGCACC


25GTGGTGTCCCAAGGC GGCGATCTCAAAAGT CCAGTCCTAGCACCT


TGGTGTCCCAAGGCC GCGATCTCAAAAGTT CAGTCCTAGCACCTC


GGTGTCCCAAGGCCA CGATCTCAAAAGTTA AGTCCTAGCACCTCC


GTGTCCCAAGGCCAG GATCTCAAAAGTTAT GTCCTAGCACCTCCA


TGTCCCAAGGCCAGC ATCTCAAAAGTTATC TCCTAGCACCTCCAA


30GTCCCAAGGCCAGCC TCTCAAAAGTTATCT CCTAGCACCTCCAAG


TCCCAAGGCCAGCCA CTCAAAAGTTATCTC CTAGCACCTCCAAGC


CCCAAGGCCAGCCAA TCAAAAGTTATCTCC TAGCACCTCCAAGCC


CCAAGGCCAGCCAAC CAAAAGTTATCTCCG AGCACCTCCAAGCCT


CAAGGCCAGCCAACA AAAAGTTATCTCCGG GCACCTCCAAGCCTG


35AAGGCCAGCCAACAC AAAGTTATCTCCGGT CACCTCCAAGCCTGA


AGGCCAGCCAACACT AAGTTATCTCCGGTC ACCTCCAAGCCTGAG


GGCCAGCCAACACTG AGTTATCTCCGGTCT CCTCCAAGCCTGAGC


GCCAGCCAACACTGG GTTATCTCCGGTCTC CTCCAAGCCTGAGCA


CCAGCCAACACTGGT TTATCTCCGGTCTCT TCCAAGCCTGAGCAA


40CAGCCAACACTGGTC TATCTCCGGTCTCTG CCAAGCCTGAGCAAG


AGCCAACACTGGTCA ATCTCCGGTCTCTGA CAAGCCTGAGCAAGA


GCCAACACTGGTCAT TCTCCGGTCTCTGAG AAGCCTGAGCAAGAT


CCAACACTGGTCATC CTCCGGTCTCTGAGG AGCCTGAGCAAGATG


CAACACTGGTCATCA TCCGGTCTCTGAGGC GCCTGAGCAAGATGA


45AACACTGGTCATCAT CCGGTCTCTGAGGCC CCTGAGCAAGATGAT


ACACTGGTCATCATG CGGTCTCTGAGGCCA CTGAGCAAGATGATT


CACTGGTCATCATGG GGTCTCTGAGGCCAG TGAGCAAGATGATTC


ACTGGTCATCATGGA GTCTCTGAGGCCAGA GAGCAAGATGATTCA


CTGGTCATCATGGAA TCTCTGAGGCCAGAA AGCAAGATGATTCAG


SOTGGTCATCATGGAAC CTCTGAGGCCAGAAA GCAAGATGATTCAGA


GGTCATCATGGAACT TCTGAGGCCAGAAAT CAAGATGATTCAGAT




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AAGATGATTCAGATG GCCAATAAGTTCGTC GAAGATTTCACAGTC


AGATGATTCAGATGG CCAATAAGTTCGTCC AAGATTTCACAGTCA


GATGATTCAGATGGC CAATAAGTTCGTCCA AGATTTCACAGTCAA


ATGATTCAGATGGCC AATAAGTTCGTCCAC GATTTCACAGTCAAA


TGATTCAGATGGCCG ATAAGTTCGTCCACA ATTTCACAGTCAAAA


GATTCAGATGGCCGG TAAGTTCGTCCACAG TTTCACAGTCAAAAT


ATTCAGATGGCCGGA AAGTTCGTCCACAGA TTCACAGTCAAAATC


TTCAGATGGCCGGAG AGTTCGTCCACAGAG TCACAGTCAAAATCG


TCAGATGGCCGGAGA GTTCGTCCACAGAGA CACAGTCAAAATCGG


10CAGATGGCCGGAGAG TTCGTCCACAGAGAC ACAGTCAAAATCGGA


AGATGGCCGGAGAGA TCGTCCACAGAGACC CAGTCAAAATCGGAG


GATGGCCGGAGAGAT CGTCCACAGAGACCT AGTCAAAATCGGAGA


ATGGCCGGAGAGATT GTCCACAGAGACCTT GTCAAAATCGGAGAT


TGGCCGGAGAGATTG TCCACAGAGACCTTG TCAAAATCGGAGATT


15GGCCGGAGAGATTGC CCACAGAGACCTTGC CAAAATCGGAGATTT


GCCGGAGAGATTGCA CACAGAGACCTTGCT AAAATCGGAGATTTT


CCGGAGAGATTGCAG ACAGAGACCTTGCTG AAATCGGAGATTTTG


CGGAGAGATTGCAGA CAGAGACCTTGCTGC AATCGGAGATTTTGG


GGAGAGATTGCAGAC AGAGACCTTGCTGCC ATCGGAGATTTTGGT


20GAGAGATTGCAGACG GAGACCTTGCTGCCC TCGGAGATTTTGGTA


AGAGATTGCAGACGG AGACCTTGCTGCCCG CGGAGATTTTGGTAT


GAGATTGCAGACGGC GACCTTGCTGCCCGG GGAGATTTTGGTATG


AGATTGCAGACGGCA ACCTTGCTGCCCGGA GAGATTTTGGTATGA


GATTGCAGACGGCAT CCTTGCTGCCCGGAA AGATTTTGGTATGAC


25ATTGCAGACGGCATG CTTGCTGCCCGGAAT GATTTTGGTATGACG


TTGCAGACGGCATGG TTGCTGCCCGGAATT ATTTTGGTATGACGC


TGCAGACGGCATGGC TGCTGCCCGGAATTG TTTTGGTATGACGCG


GCAGACGGCATGGCA GCTGCCCGGAATTGC TTTGGTATGACGCGA


CAGACGGCATGGCAT CTGCCCGGAATTGCA TTGGTATGACGCGAG


30AGACGGCATGGCATA TGCCCGGAATTGCAT TGGTATGACGCGAGA


GACGGCATGGCATAC GCCCGGAATTGCATG GGTATGACGCGAGAT


ACGGCATGGCATACC CCCGGAATTGCATGG GTATGACGCGAGATA


CGGCATGGCATACCT CCGGAATTGCATGGT TATGACGCGAGATAT


GGCATGGCATACCTC CGGAATTGCATGGTA ATGACGCGAGATATC


35GCATGGCATACCTCA GGAATTGCATGGTAG TGACGCGAGATATCT


CATGGCATACCTCAA GAATTGCATGGTAGC GACGCGAGATATCTA


ATGGCATACCTCAAC AATTGCATGGTAGCC ACGCGAGATATCTAT


TGGCATACCTCAACG ATTGCATGGTAGCCG CGCGAGATATCTATG


GGCATACCTCAACGC TTGCATGGTAGCCGA GCGAGATATCTATGA


40GCATACCTCAACGCC TGCATGGTAGCCGAA CGAGATATCTATGAG


CATACCTCAACGCCA GCATGGTAGCCGAAG GAGATATCTATGAGA


ATACCTCAACGCCAA CATGGTAGCCGAAGA AGATATCTATGAGAC


TACCTCAACGCCAAT ATGGTAGCCGAAGAT GATATCTATGAGACA


ACCTCAACGCCAATA TGGTAGCCGAAGATT ATATCTATGAGACAG


45CCTCAACGCCAATAA GGTAGCCGAAGATTT TATCTATGAGACAGA


CTCAACGCCAATAAG GTAGCCGAAGATTTC ATCTATGAGACAGAC


TCAACGCCAATAAGT TAGCCGAAGATTTCA TCTATGAGACAGACT


CAACGCCAATAAGTT AGCCGAAGATTTCAC CTATGAGACAGACTA


AACGCCAATAAGTTC GCCGAAGATTTCACA TATGAGACAGACTAT


50ACGCCAATAAGTTCG CCGAAGATTTCACAG ATGAGACAGACTATT


CGCCAATAAGTTCGT CGAAGATTTCACAGT TGAGACAGACTATTA




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-84-
GAGACAGACTATTAC ATGTCTCCTGAGTCC TGGTCCTTCGGGGTC


AGACAGACTATTACC TGTCTCCTGAGTCCC GGTCCTTCGGGGTCG


GACAGACTATTACCG GTCTCCTGAGTCCCT GTCCTTCGGGGTCGT


ACAGACTATTACCGG TCTCCTGAGTCCCTC TCCTTCGGGGTCGTC


CAGACTATTACCGGA CTCCTGAGTCCCTCA CCTTCGGGGTCGTCC


AGACTATTACCGGAA TCCTGAGTCCCTCAA CTTCGGGGTCGTCCT


GACTATTACCGGAAA CCTGAGTCCCTCAAG TTCGGGGTCGTCCTC


ACTATTACCGGAAAG CTGAGTCCCTCAAGG TCGGGGTCGTCCTCT


CTATTACCGGAAAGG TGAGTCCCTCAAGGA CGGGGTCGTCCTCTG


10TATTACCGGAAAGGA GAGTCCCTCAAGGAT GGGGTCGTCCTCTGG


ATTACCGGAAAGGAG AGTCCCTCAAGGATG GGGTCGTCCTCTGGG


TTACCGGAAAGGAGG GTCCCTCAAGGATGG GGTCGTCCTCTGGGA


TACCGGAAAGGAGGC TCCCTCAAGGATGGA GTCGTCCTCTGGGAG


ACCGGAAAGGAGGCA CCCTCAAGGATGGAG TCGTCCTCTGGGAGA


15CCGGAAAGGAGGCAA CCTCAAGGATGGAGT CGTCCTCTGGGAGAT


CGGAAAGGAGGCAAA CTCAAGGATGGAGTC GTCCTCTGGGAGATC


GGAAAGGAGGCAAAG TCAAGGATGGAGTCT TCCTCTGGGAGATCG


GAAAGGAGGCAAAGG CAAGGATGGAGTCTT CCTCTGGGAGATCGC


AAAGGAGGCAAAGGG AAGGATGGAGTCTTC CTCTGGGAGATCGCC


20AAGGAGGCAAAGGGC AGGATGGAGTCTTCA TCTGGGAGATCGCCA


AGGAGGCAAAGGGCT GGATGGAGTCTTCAC CTGGGAGATCGCCAC


GGAGGCAAAGGGCTG GATGGAGTCTTCACC TGGGAGATCGCCACA


GAGGCAAAGGGCTGC ATGGAGTCTTCACCA GGGAGATCGCCACAC


AGGCAAAGGGCTGCT TGGAGTCTTCACCAC GGAGATCGCCACACT


25GGCAAAGGGCTGCTG GGAGTCTTCACCACT GAGATCGCCACACTG


GCAAAGGGCTGCTGC GAGTCTTCACCACTT AGATCGCCACACTGG


CAAAGGGCTGCTGCC AGTCTTCACCACTTA GATCGCCACACTGGC


AAAGGGCTGCTGCCC GTCTTCACCACTTAC ATCGCCACACTGGCC


AAGGGCTGCTGCCCG TCTTCACCACTTACT TCGCCACACTGGCCG


30AGGGCTGCTGCCCGT CTTCACCACTTACTC CGCCACACTGGCCGA


GGGCTGCTGCCCGTG TTCACCACTTACTCG GCCACACTGGCCGAG


GGCTGCTGCCCGTGC TCACCACTTACTCGG CCACACTGGCCGAGC


GCTGCTGCCCGTGCG CACCACTTACTCGGA CACACTGGCCGAGCA


CTGCTGCCCGTGCGC ACCACTTACTCGGAC ACACTGGCCGAGCAG


35TGCTGCCCGTGCGCT CCACTTACTCGGACG CACTGGCCGAGCAGC


GCTGCCCGTGCGCTG CACTTACTCGGACGT ACTGGCCGAGCAGCC


CTGCCCGTGCGCTGG ACTTACTCGGACGTC CTGGCCGAGCAGCCC


TGCCCGTGCGCTGGA CTTACTCGGACGTCT TGGCCGAGCAGCCCT


GCCCGTGCGCTGGAT TTACTCGGACGTCTG GGCCGAGCAGCCCTA


40CCCGTGCGCTGGATG TACTCGGACGTCTGG GCCGAGCAGCCCTAC


CCGTGCGCTGGATGT ACTCGGACGTCTGGT CCGAGCAGCCCTACC


CGTGCGCTGGATGTC CTCGGACGTCTGGTC CGAGCAGCCCTACCA


GTGCGCTGGATGTCT TCGGACGTCTGGTCC GAGCAGCCCTACCAG


TGCGCTGGATGTCTC CGGACGTCTGGTCCT AGCAGCCCTACCAGG


45GCGCTGGATGTCTCC GGACGTCTGGTCCTT GCAGCCCTACCAGGG


CGCTGGATGTCTCCT GACGTCTGGTCCTTC CAGCCCTACCAGGGC


GCTGGATGTCTCCTG ACGTCTGGTCCTTCG AGCCCTACCAGGGCT


CTGGATGTCTCCTGA CGTCTGGTCCTTCGG GCCCTACCAGGGCTT


TGGATGTCTCCTGAG GTCTGGTCCTTCGGG CCCTACCAGGGCTTG


50GGATGTCTCCTGAGT TCTGGTCCTTCGGGG CCTACCAGGGCTTGT


GATGTCTCCTGAGTC CTGGTCCTTCGGGGT CTACCAGGGCTTGTC




CA 02376284 2001-12-05
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-85-
TACCAGGGCTTGTCC CTTCTGGACAAGCCA ATGTGCTGGCAGTAT


ACCAGGGCTTGTCCA TTCTGGACAAGCCAG TGTGCTGGCAGTATA


CCAGGGCTTGTCCAA TCTGGACAAGCCAGA GTGCTGGCAGTATAA


CAGGGCTTGTCCAAC CTGGACAAGCCAGAC TGCTGGCAGTATAAC


AGGGCTTGTCCAACG TGGACAAGCCAGACA GCTGGCAGTATAACC


GGGCTTGTCCAACGA GGACAAGCCAGACAA CTGGCAGTATAACCC


GGCTTGTCCAACGAG GACAAGCCAGACAAC TGGCAGTATAACCCC


GCTTGTCCAACGAGC ACAAGCCAGACAACT GGCAGTATAACCCCA


CTTGTCCAACGAGCA CAAGCCAGACAACTG GCAGTATAACCCCAA


10TTGTCCAACGAGCAA AAGCCAGACAACTGT CAGTATAACCCCAAG


TGTCCAACGAGCAAG AGCCAGACAACTGTC AGTATAACCCCAAGA


GTCCAACGAGCAAGT GCCAGACAACTGTCC GTATAACCCCAAGAT


TCCAACGAGCAAGTC CCAGACAACTGTCCT TATAACCCCAAGATG


CCAACGAGCAAGTCC CAGACAACTGTCCTG ATAACCCCAAGATGA


15CAACGAGCAAGTCCT AGACAACTGTCCTGA TAACCCCAAGATGAG


AACGAGCAAGTCCTT GACAACTGTCCTGAC AACCCCAAGATGAGG


ACGAGCAAGTCCTTC ACAACTGTCCTGACA ACCCCAAGATGAGGC


CGAGCAAGTCCTTCG CAACTGTCCTGACAT CCCCAAGATGAGGCC


GAGCAAGTCCTTCGC AACTGTCCTGACATG CCCAAGATGAGGCCT


20AGCAAGTCCTTCGCT ACTGTCCTGACATGC CCAAGATGAGGCCTT


GCAAGTCCTTCGCTT CTGTCCTGACATGCT CAAGATGAGGCCTTC


CAAGTCCTTCGCTTC TGTCCTGACATGCTG AAGATGAGGCCTTCC


AAGTCCTTCGCTTCG GTCCTGACATGCTGT AGATGAGGCCTTCCT


AGTCCTTCGCTTCGT TCCTGACATGCTGTT GATGAGGCCTTCCTT


25GTCCTTCGCTTCGTC CCTGACATGCTGTTT ATGAGGCCTTCCTTC


TCCTTCGCTTCGTCA CTGACATGCTGTTTG TGAGGCCTTCCTTCC


CCTTCGCTTCGTCAT TGACATGCTGTTTGA GAGGCCTTCCTTCCT


CTTCGCTTCGTCATG GACATGCTGTTTGAA AGGCCTTCCTTCCTG


TTCGCTTCGTCATGG ACATGCTGTTTGAAC GGCCTTCCTTCCTGG


30TCGCTTCGTCATGGA CATGCTGTTTGAACT GCCTTCCTTCCTGGA


CGCTTCGTCATGGAG ATGCTGTTTGAACTG CCTTCCTTCCTGGAG


GCTTCGTCATGGAGG TGCTGTTTGAACTGA CTTCCTTCCTGGAGA


CTTCGTCATGGAGGG GCTGTTTGAACTGAT TTCCTTCCTGGAGAT


TTCGTCATGGAGGGC CTGTTTGAACTGATG TCCTTCCTGGAGATC


35TCGTCATGGAGGGCG TGTTTGAACTGATGC CCTTCCTGGAGATCA


CGTCATGGAGGGCGG GTTTGAACTGATGCG CTTCCTGGAGATCAT


GTCATGGAGGGCGGC TTTGAACTGATGCGC TTCCTGGAGATCATC


TCATGGAGGGCGGCC TTGAACTGATGCGCA TCCTGGAGATCATCA


CATGGAGGGCGGCCT TGAACTGATGCGCAT CCTGGAGATCATCAG


40ATGGAGGGCGGCCTT GAACTGATGCGCATG CTGGAGATCATCAGC


TGGAGGGCGGCCTTC AACTGATGCGCATGT TGGAGATCATCAGCA


GGAGGGCGGCCTTCT ACTGATGCGCATGTG GGAGATCATCAGCAG


GAGGGCGGCCTTCTG CTGATGCGCATGTGC GAGATCATCAGCAGC


AGGGCGGCCTTCTGG TGATGCGCATGTGCT AGATCATCAGCAGCA


45GGGCGGCCTTCTGGA GATGCGCATGTGCTG GATCATCAGCAGCAT


GGCGGCCTTCTGGAC ATGCGCATGTGCTGG ATCATCAGCAGCATC


GCGGCCTTCTGGACA TGCGCATGTGCTGGC TCATCAGCAGCATCA


CGGCCTTCTGGACAA GCGCATGTGCTGGCA CATCAGCAGCATCAA


GGCCTTCTGGACAAG CGCATGTGCTGGCAG ATCAGCAGCATCAAA


50GCCTTCTGGACAAGC GCATGTGCTGGCAGT TCAGCAGCATCAAAG


CCTTCTGGACAAGCC CATGTGCTGGCAGTA CAGCAGCATCAAAGA




CA 02376284 2001-12-05
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-86-
AGCAGCATCAAAGAG TACAGCGAGGAGAAC GAGAACATGGAGAGC


GCAGCATCAAAGAGG ACAGCGAGGAGAACA AGAACATGGAGAGCG


CAGCATCAAAGAGGA CAGCGAGGAGAACAA GAACATGGAGAGCGT


AGCATCAAAGAGGAG AGCGAGGAGAACAAG AACATGGAGAGCGTC


GCATCAAAGAGGAGA GCGAGGAGAACAAGC ACATGGAGAGCGTCC


CATCAAAGAGGAGAT CGAGGAGAACAAGCT CATGGAGAGCGTCCC


ATCAAAGAGGAGATG GAGGAGAACAAGCTG ATGGAGAGCGTCCCC


TCAAAGAGGAGATGG AGGAGAACAAGCTGC TGGAGAGCGTCCCCC


CAAAGAGGAGATGGA GGAGAACAAGCTGCC GGAGAGCGTCCCCCT


10AAAGAGGAGATGGAG GAGAACAAGCTGCCC GAGAGCGTCCCCCTG


AAGAGGAGATGGAGC AGAACAAGCTGCCCG AGAGCGTCCCCCTGG


AGAGGAGATGGAGCC GAACAAGCTGCCCGA GAGCGTCCCCCTGGA


GAGGAGATGGAGCCT AACAAGCTGCCCGAG AGCGTCCCCCTGGAC


AGGAGATGGAGCCTG ACAAGCTGCCCGAGC GCGTCCCCCTGGACC


15GGAGATGGAGCCTGG CAAGCTGCCCGAGCC CGTCCCCCTGGACCC


GAGATGGAGCCTGGC AAGCTGCCCGAGCCG GTCCCCCTGGACCCC


AGATGGAGCCTGGCT AGCTGCCCGAGCCGG TCCCCCTGGACCCCT


GATGGAGCCTGGCTT GCTGCCCGAGCCGGA CCCCCTGGACCCCTC


ATGGAGCCTGGCTTC CTGCCCGAGCCGGAG CCCCTGGACCCCTCG


20TGGAGCCTGGCTTCC TGCCCGAGCCGGAGG CCCTGGACCCCTCGG


GGAGCCTGGCTTCCG GCCCGAGCCGGAGGA CCTGGACCCCTCGGC


GAGCCTGGCTTCCGG CCCGAGCCGGAGGAG CTGGACCCCTCGGCC


AGCCTGGCTTCCGGG CCGAGCCGGAGGAGC TGGACCCCTCGGCCT


GCCTGGCTTCCGGGA CGAGCCGGAGGAGCT GGACCCCTCGGCCTC


25CCTGGCTTCCGGGAG GAGCCGGAGGAGCTG GACCCCTCGGCCTCC


CTGGCTTCCGGGAGG AGCCGGAGGAGCTGG ACCCCTCGGCCTCCT


TGGCTTCCGGGAGGT GCCGGAGGAGCTGGA CCCCTCGGCCTCCTC


GGCTTCCGGGAGGTC CCGGAGGAGCTGGAC CCCTCGGCCTCCTCG


GCTTCCGGGAGGTCT CGGAGGAGCTGGACC CCTCGGCCTCCTCGT


30CTTCCGGGAGGTCTC GGAGGAGCTGGACCT CTCGGCCTCCTCGTC


TTCCGGGAGGTCTCC GAGGAGCTGGACCTG TCGGCCTCCTCGTCC


TCCGGGAGGTCTCCT AGGAGCTGGACCTGG CGGCCTCCTCGTCCT


CCGGGAGGTCTCCTT GGAGCTGGACCTGGA GGCCTCCTCGTCCTC


CGGGAGGTCTCCTTC GAGCTGGACCTGGAG GCCTCCTCGTCCTCC


35GGGAGGTCTCCTTCT AGCTGGACCTGGAGC CCTCCTCGTCCTCCC


GGAGGTCTCCTTCTA GCTGGACCTGGAGCC CTCCTCGTCCTCCCT


GAGGTCTCCTTCTAC CTGGACCTGGAGCCA TCCTCGTCCTCCCTG


AGGTCTCCTTCTACT TGGACCTGGAGCCAG CCTCGTCCTCCCTGC


GGTCTCCTTCTACTA GGACCTGGAGCCAGA CTCGTCCTCCCTGCC


40GTCTCCTTCTACTAC GACCTGGAGCCAGAG TCGTCCTCCCTGCCA


TCTCCTTCTACTACA ACCTGGAGCCAGAGA CGTCCTCCCTGCCAC


CTCCTTCTACTACAG CCTGGAGCCAGAGAA GTCCTCCCTGCCACT


TCCTTCTACTACAGC CTGGAGCCAGAGAAC TCCTCCCTGCCACTG


CCTTCTACTACAGCG TGGAGCCAGAGAACA CCTCCCTGCCACTGC


45CTTCTACTACAGCGA GGAGCCAGAGAACAT CTCCCTGCCACTGCC


TTCTACTACAGCGAG GAGCCAGAGAACATG TCCCTGCCACTGCCC


TCTACTACAGCGAGG AGCCAGAGAACATGG CCCTGCCACTGCCCG


CTACTACAGCGAGGA GCCAGAGAACATGGA CCTGCCACTGCCCGA


TACTACAGCGAGGAG CCAGAGAACATGGAG CTGCCACTGCCCGAC


SOACTACAGCGAGGAGA CAGAGAACATGGAGA TGCCACTGCCCGACA


CTACAGCGAGGAGAA AGAGAACATGGAGAG GCCACTGCCCGACAG




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
_g7_
CCACTGCCCGACAGA GGGGTGCTGGTCCTC ATGAACGGGGGCCGC


CACTGCCCGACAGAC GGGTGCTGGTCCTCC TGAACGGGGGCCGCA


ACTGCCCGACAGACA GGTGCTGGTCCTCCG GAACGGGGGCCGCAA


CTGCCCGACAGACAC GTGCTGGTCCTCCGC AACGGGGGCCGCAAG


TGCCCGACAGACACT TGCTGGTCCTCCGCG ACGGGGGCCGCAAGA


GCCCGACAGACACTC GCTGGTCCTCCGCGC CGGGGGCCGCAAGAA


CCCGACAGACACTCA CTGGTCCTCCGCGCC GGGGGCCGCAAGAAC


CCGACAGACACTCAG TGGTCCTCCGCGCCA GGGGCCGCAAGAACG


CGACAGACACTCAGG GGTCCTCCGCGCCAG GGGCCGCAAGAACGA


10GACAGACACTCAGGA GTCCTCCGCGCCAGC GGCCGCAAGAACGAG


ACAGACACTCAGGAC TCCTCCGCGCCAGCT GCCGCAAGAACGAGC


CAGACACTCAGGACA CCTCCGCGCCAGCTT CCGCAAGAACGAGCG


AGACACTCAGGACAC CTCCGCGCCAGCTTC CGCAAGAACGAGCGG


GACACTCAGGACACA TCCGCGCCAGCTTCG GCAAGAACGAGCGGG


15ACACTCAGGACACAA CCGCGCCAGCTTCGA CAAGAACGAGCGGGC


CACTCAGGACACAAG CGCGCCAGCTTCGAC AAGAACGAGCGGGCC


ACTCAGGACACAAGG GCGCCAGCTTCGACG AGAACGAGCGGGCCT


CTCAGGACACAAGGC CGCCAGCTTCGACGA GAACGAGCGGGCCTT


TCAGGACACAAGGCC GCCAGCTTCGACGAG AACGAGCGGGCCTTG


20CAGGACACAAGGCCG CCAGCTTCGACGAGA ACGAGCGGGCCTTGC


AGGACACAAGGCCGA CAGCTTCGACGAGAG CGAGCGGGCCTTGCC


GGACACAAGGCCGAG AGCTTCGACGAGAGA GAGCGGGCCTTGCCG


GACACAAGGCCGAGA GCTTCGACGAGAGAC AGCGGGCCTTGCCGC


ACACAAGGCCGAGAA CTTCGACGAGAGACA GCGGGCCTTGCCGCT


25CACAAGGCCGAGAAC TTCGACGAGAGACAG CGGGCCTTGCCGCTG


ACAAGGCCGAGAACG TCGACGAGAGACAGC GGGCCTTGCCGCTGC


CAAGGCCGAGAACGG CGACGAGAGACAGCC GGCCTTGCCGCTGCC


AAGGCCGAGAACGGC GACGAGAGACAGCCT GCCTTGCCGCTGCCC


AGGCCGAGAACGGCC ACGAGAGACAGCCTT CCTTGCCGCTGCCCC


30GGCCGAGAACGGCCC CGAGAGACAGCCTTA CTTGCCGCTGCCCCA


GCCGAGAACGGCCCC GAGAGACAGCCTTAC TTGCCGCTGCCCCAG


CCGAGAACGGCCCCG AGAGACAGCCTTACG TGCCGCTGCCCCAGT


CGAGAACGGCCCCGG GAGACAGCCTTACGC GCCGCTGCCCCAGTC


GAGAACGGCCCCGGC AGACAGCCTTACGCC CCGCTGCCCCAGTCT


35AGAACGGCCCCGGCC GACAGCCTTACGCCC CGCTGCCCCAGTCTT


GAACGGCCCCGGCCC ACAGCCTTACGCCCA GCTGCCCCAGTCTTC


AACGGCCCCGGCCCT CAGCCTTACGCCCAC CTGCCCCAGTCTTCG


ACGGCCCCGGCCCTG AGCCTTACGCCCACA TGCCCCAGTCTTCGA


CGGCCCCGGCCCTGG GCCTTACGCCCACAT GCCCCAGTCTTCGAC


40GGCCCCGGCCCTGGG CCTTACGCCCACATG CCCCAGTCTTCGACC


GCCCCGGCCCTGGGG CTTACGCCCACATGA CCCAGTCTTCGACCT


CCCCGGCCCTGGGGT TTACGCCCACATGAA CCAGTCTTCGACCTG


CCCGGCCCTGGGGTG TACGCCCACATGAAC CAGTCTTCGACCTGC


CCGGCCCTGGGGTGC ACGCCCACATGAACG AGTCTTCGACCTGCT


45CGGCCCTGGGGTGCT CGCCCACATGAACGG GTCTTCGACCTGCTG


GGCCCTGGGGTGCTG GCCCACATGAACGGG TCTTCGACCTGCTGA


GCCCTGGGGTGCTGG CCCACATGAACGGGG CTTCGACCTGCTGAT


CCCTGGGGTGCTGGT CCACATGAACGGGGG TTCGACCTGCTGATC


CCTGGGGTGCTGGTC CACATGAACGGGGGC TCGACCTGCTGATCC


50CTGGGGTGCTGGTCC ACATGAACGGGGGCC CGACCTGCTGATCCT


TGGGGTGCTGGTCCT CATGAACGGGGGCCG GACCTGCTGATCCTT




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
_$g_
ACCTGCTGATCCTTG GCGCAGCGGGGTGGG TCCTGTACCTCAGTG


CCTGCTGATCCTTGG CGCAGCGGGGTGGGG CCTGTACCTCAGTGG


CTGCTGATCCTTGGA GCAGCGGGGTGGGGG CTGTACCTCAGTGGA


TGCTGATCCTTGGAT CAGCGGGGTGGGGGG TGTACCTCAGTGGAT


GCTGATCCTTGGATC AGCGGGGTGGGGGGG GTACCTCAGTGGATC


CTGATCCTTGGATCC GCGGGGTGGGGGGGG TACCTCAGTGGATCT


TGATCCTTGGATCCT CGGGGTGGGGGGGGA ACCTCAGTGGATCTT


GATCCTTGGATCCTG GGGGTGGGGGGGGAG CCTCAGTGGATCTTC


ATCCTTGGATCCTGA GGGTGGGGGGGGAGA CTCAGTGGATCTTCA


10TCCTTGGATCCTGAA GGTGGGGGGGGAGAG TCAGTGGATCTTCAG


CCTTGGATCCTGAAT GTGGGGGGGGAGAGA CAGTGGATCTTCAGT


CTTGGATCCTGAATC TGGGGGGGGAGAGAG AGTGGATCTTCAGTT


TTGGATCCTGAATCT GGGGGGGGAGAGAGA GTGGATCTTCAGTTC


TGGATCCTGAATCTG GGGGGGGAGAGAGAG TGGATCTTCAGTTCT


15GGATCCTGAATCTGT GGGGGGAGAGAGAGT GGATCTTCAGTTCTG


GATCCTGAATCTGTG GGGGGAGAGAGAGTT GATCTTCAGTTCTGC


ATCCTGAATCTGTGC GGGGAGAGAGAGTTT ATCTTCAGTTCTGCC


TCCTGAATCTGTGCA GGGAGAGAGAGTTTT TCTTCAGTTCTGCCC


CCTGAATCTGTGCAA GGAGAGAGAGTTTTA CTTCAGTTCTGCCCT


20CTGAATCTGTGCAAA GAGAGAGAGTTTTAA TTCAGTTCTGCCCTT


TGAATCTGTGCAAAC AGAGAGAGTTTTAAC TCAGTTCTGCCCTTG


GAATCTGTGCAAACA GAGAGAGTTTTAACA CAGTTCTGCCCTTGC


AATCTGTGCAAACAG AGAGAGTTTTAACAA AGTTCTGCCCTTGCT


ATCTGTGCAAACAGT GAGAGTTTTAACAAT GTTCTGCCCTTGCTG
~


25TCTGTGCAAACAGTA AGAGTTTTAACAATC TTCTGCCCTTGCTGC


CTGTGCAAACAGTAA GAGTTTTAACAATCC TCTGCCCTTGCTGCC


TGTGCAAACAGTAAC AGTTTTAACAATCCA CTGCCCTTGCTGCCC


GTGCAAACAGTAACG GTTTTAACAATCCAT TGCCCTTGCTGCCCG


TGCAAACAGTAACGT TTTTAACAATCCATT GCCCTTGCTGCCCGC


30GCAAACAGTAACGTG TTTAACAATCCATTC CCCTTGCTGCCCGCG


CAAACAGTAACGTGT TTAACAATCCATTCA CCTTGCTGCCCGCGG


AAACAGTAACGTGTG TAACAATCCATTCAC CTTGCTGCCCGCGGG


AACAGTAACGTGTGC AACAATCCATTCACA TTGCTGCCCGCGGGA


ACAGTAACGTGTGCG ACAATCCATTCACAA TGCTGCCCGCGGGAG


35CAGTAACGTGTGCGC CAATCCATTCACAAG GCTGCCCGCGGGAGA


AGTAACGTGTGCGCA AATCCATTCACAAGC CTGCCCGCGGGAGAC


GTAACGTGTGCGCAC ATCCATTCACAAGCC TGCCCGCGGGAGACA


TAACGTGTGCGCACG TCCATTCACAAGCCT GCCCGCGGGAGACAG


AACGTGTGCGCACGC CCATTCACAAGCCTC . CCCGCGGGAGACAGC


40ACGTGTGCGCACGCG CATTCACAAGCCTCC CCGCGGGAGACAGCT


CGTGTGCGCACGCGC ATTCACAAGCCTCCT CGCGGGAGACAGCTT


GTGTGCGCACGCGCA TTCACAAGCCTCCTG GCGGGAGACAGCTTC


TGTGCGCACGCGCAG TCACAAGCCTCCTGT CGGGAGACAGCTTCT


GTGCGCACGCGCAGC CACAAGCCTCCTGTA GGGAGACAGCTTCTC


45TGCGCACGCGCAGCG ACAAGCCTCCTGTAC GGAGACAGCTTCTCT


GCGCACGCGCAGCGG CAAGCCTCCTGTACC GAGACAGCTTCTCTG


CGCACGCGCAGCGGG AAGCCTCCTGTACCT AGACAGCTTCTCTGC


GCACGCGCAGCGGGG AGCCTCCTGTACCTC GACAGCTTCTCTGCA


CACGCGCAGCGGGGT GCCTCCTGTACCTCA ACAGCTTCTCTGCAG


50ACGCGCAGCGGGGTG CCTCCTGTACCTCAG CAGCTTCTCTGCAGT


CGCGCAGCGGGGTGG CTCCTGTACCTCAGT AGCTTCTCTGCAGTA




CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-89-
GCTTCTCTGCAGTAA CAGCTTTTTATTCCC CTTAATGACAACACT


CTTCTCTGCAGTAAA AGCTTTTTATTCCCT TTAATGACAACACTT


TTCTCTGCAGTAAAA GCTTTTTATTCCCTG TAATGACAACACTTA


TCTCTGCAGTAAAAC CTTTTTATTCCCTGC AATGACAACACTTAA


CTCTGCAGTAAAACA TTTTTATTCCCTGCC ATGACAACACTTAAT


TCTGCAGTAAAACAC TTTTATTCCCTGCCC TGACAACACTTAATA


CTGCAGTAAAACACA TTTATTCCCTGCCCA GACAACACTTAATAG


TGCAGTAAAACACAT TTATTCCCTGCCCAA ACAACACTTAATAGC


GCAGTAAAACACATT TATTCCCTGCCCAAA CAACACTTAATAGCA


10CAGTAAAACACATTT ATTCCCTGCCCAAAC AACACTTAATAGCAA


AGTAAAACACATTTG TTCCCTGCCCAAACC ACACTTAATAGCAAC


GTAAAACACATTTGG TCCCTGCCCAAACCC CACTTAATAGCAACA


TAAAACACATTTGGG CCCTGCCCAAACCCT ACTTAATAGCAACAG


AAAACACATTTGGGA CCTGCCCAAACCCTT CTTAATAGCAACAGA


15AAACACATTTGGGAT CTGCCCAAACCCTTA TTAATAGCAACAGAG


AACACATTTGGGATG TGCCCAAACCCTTAA TAATAGCAACAGAGC


ACACATTTGGGATGT GCCCAAACCCTTAAC AATAGCAACAGAGCA


CACATTTGGGATGTT CCCAAACCCTTAACT ATAGCAACAGAGCAC


ACATTTGGGATGTTC CCAAACCCTTAACTG TAGCAACAGAGCACT


20CATTTGGGATGTTCC CAAACCCTTAACTGA AGCAACAGAGCACTT


ATTTGGGATGTTCCT AAACCCTTAACTGAC GCAACAGAGCACTTG


TTTGGGATGTTCCTT AACCCTTAACTGACA CAACAGAGCACTTGA


TTGGGATGTTCCTTT ACCCTTAACTGACAT AACAGAGCACTTGAG


TGGGATGTTCCTTTT CCCTTAACTGACATG ACAGAGCACTTGAGA


25GGGATGTTCCTTTTT CCTTAACTGACATGG CAGAGCACTTGAGAA


GGATGTTCCTTTTTT CTTAACTGACATGGG AGAGCACTTGAGAAC


GATGTTCCTTTTTTC TTAACTGACATGGGC GAGCACTTGAGAACC


ATGTTCCTTTTTTCA TAACTGACATGGGCC AGCACTTGAGAACCA


TGTTCCTTTTTTCAA AACTGACATGGGCCT GCACTTGAGAACCAG


30GTTCCTTTTTTCAAT ACTGACATGGGCCTT CACTTGAGAACCAGT


TTCCTTTTTTCAATA CTGACATGGGCCTTT ACTTGAGAACCAGTC


TCCTTTTTTCAATAT TGACATGGGCCTTTA CTTGAGAACCAGTCT


CCTTTTTTCAATATG GACATGGGCCTTTAA TTGAGAACCAGTCTC


CTTTTTTCAATATGC ACATGGGCCTTTAAG TGAGAACCAGTCTCC


35TTTTTTCAATATGCA CATGGGCCTTTAAGA GAGAACCAGTCTCCT


TTTTTCAATATGCAA ATGGGCCTTTAAGAA AGAACCAGTCTCCTC


TTTTCAATATGCAAG TGGGCCTTTAAGAAC GAACCAGTCTCCTCA


TTTCAATATGCAAGC GGGCCTTTAAGAACC AACCAGTCTCCTCAC


TTCAATATGCAAGCA GGCCTTTAAGAACCT ACCAGTCTCCTCACT


40TCAATATGCAAGCAG GCCTTTAAGAACCTT CCAGTCTCCTCACTC


CAATATGCAAGCAGC CCTTTAAGAACCTTA CAGTCTCCTCACTCT


AATATGCAAGCAGCT CTTTAAGAACCTTAA AGTCTCCTCACTCTG


ATATGCAAGCAGCTT TTTAAGAACCTTAAT GTCTCCTCACTCTGT


TATGCAAGCAGCTTT TTAAGAACCTTAATG TCTCCTCACTCTGTC


45ATGCAAGCAGCTTTT TAAGAACCTTAATGA CTCCTCACTCTGTCC


TGCAAGCAGCTTTTT AAGAACCTTAATGAC TCCTCACTCTGTCCC


GCAAGCAGCTTTTTA AGAACCTTAATGACA CCTCACTCTGTCCCT


CAAGCAGCTTTTTAT GAACCTTAATGACAA CTCACTCTGTCCCTG


AAGCAGCTTTTTATT AACCTTAATGACAAC TCACTCTGTCCCTGT


50AGCAGCTTTTTATTC ACCTTAATGACAACA CACTCTGTCCCTGTC


GCAGCTTTTTATTCC CCTTAATGACAACAC ACTCTGTCCCTGTCC




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CTCTGTCCCTGTCCT AACGGAAAAATAATT TGAGGAAGTGGCTGT


TCTGTCCCTGTCCTT ACGGAAA.AATAATTG GAGGAAGTGGCTGTC


CTGTCCCTGTCCTTC CGGAAAA.ATAATTGC AGGAAGTGGCTGTCC


TGTCCCTGTCCTTCC GGAAAAATAATTGCC GGAAGTGGCTGTCCC


S GTCCCTGTCCTTCCC GAAAAATAATTGCCA GAAGTGGCTGTCCCT


TCCCTGTCCTTCCCT AAAAATAATTGCCAC AAGTGGCTGTCCCTG


CCCTGTCCTTCCCTG AAAATAATTGCCACA AGTGGCTGTCCCTGT


CCTGTCCTTCCCTGT AAATAATTGCCACAA GTGGCTGTCCCTGTG


CTGTCCTTCCCTGTT AATAATTGCCACAAG TGGCTGTCCCTGTGG


10TGTCCTTCCCTGTTC ATAATTGCCACAAGT GGCTGTCCCTGTGGC


GTCCTTCCCTGTTCT TAATTGCCACAAGTC GCTGTCCCTGTGGCC


TCCTTCCCTGTTCTC AATTGCCACAAGTCC CTGTCCCTGTGGCCC


CCTTCCCTGTTCTCC ATTGCCACAAGTCCA TGTCCCTGTGGCCCC


CTTCCCTGTTCTCCC TTGCCACAAGTCCAG GTCCCTGTGGCCCCA


15TTCCCTGTTCTCCCT TGCCACAAGTCCAGC TCCCTGTGGCCCCAT


TCCCTGTTCTCCCTT GCCACAAGTCCAGCT CCCTGTGGCCCCATC


CCCTGTTCTCCCTTT CCACAAGTCCAGCTG CCTGTGGCCCCATCC


CCTGTTCTCCCTTTC CACAAGTCCAGCTGG CTGTGGCCCCATCCA


CTGTTCTCCCTTTCT ACAAGTCCAGCTGGG TGTGGCCCCATCCAA


20TGTTCTCCCTTTCTC CAAGTCCAGCTGGGA GTGGCCCCATCCAAC


GTTCTCCCTTTCTCT AAGTCCAGCTGGGAA TGGCCCCATCCAACC


TTCTCCCTTTCTCTC AGTCCAGCTGGGAAG GGCCCCATCCAACCA


TCTCCCTTTCTCTCT GTCCAGCTGGGAAGC GCCCCATCCAACCAC


CTCCCTTTCTCTCTC TCCAGCTGGGAAGCC CCCCATCCAACCACT


25TCCCTTTCTCTCTCC CCAGCTGGGAAGCCC CCCATCCAACCACTG


CCCTTTCTCTCTCCT CAGCTGGGAAGCCCT CCATCCAACCACTGT


CCTTTCTCTCTCCTC AGCTGGGAAGCCCTT CATCCAACCACTGTA


CTTTCTCTCTCCTCT GCTGGGAAGCCCTTT ATCCAACCACTGTAC


TTTCTCTCTCCTCTC CTGGGAAGCCCTTTT TCCAACCACTGTACA


30TTCTCTCTCCTCTCT TGGGAAGCCCTTTTT CCAACCACTGTACAC


TCTCTCTCCTCTCTG GGGAAGCCCTTTTTA CAACCACTGTACACA


CTCTCTCCTCTCTGC GGAAGCCCTTTTTAT AACCACTGTACACAC


TCTCTCCTCTCTGCT GAAGCCCTTTTTATC ACCACTGTACACACC


CTCTCCTCTCTGCTT AAGCCCTTTTTATCA CCACTGTACACACCC


35TCTCCTCTCTGCTTC AGCCCTTTTTATCAG CACTGTACACACCCG


CTCCTCTCTGCTTCA GCCCTTTTTATCAGT ACTGTACACACCCGC


TCCTCTCTGCTTCAT CCCTTTTTATCAGTT CTGTACACACCCGCC


CCTCTCTGCTTCATA CCTTTTTATCAGTTT TGTACACACCCGCCT


CTCTCTGCTTCATAA CTTTTTATCAGTTTG GTACACACCCGCCTG


40TCTCTGCTTCATAAC TTTTTATCAGTTTGA TACACACCCGCCTGA


CTCTGCTTCATAACG TTTTATCAGTTTGAG ACACACCCGCCTGAC


TCTGCTTCATAACGG TTTATCAGTTTGAGG CACACCCGCCTGACA


CTGCTTCATAACGGA TTATCAGTTTGAGGA ACACCCGCCTGACAC


TGCTTCATAACGGAA TATCAGTTTGAGGAA CACCCGCCTGACACC


45GCTTCATAACGGAAA ATCAGTTTGAGGAAG ACCCGCCTGACACCG


CTTCATAACGGAAAA TCAGTTTGAGGAAGT CCCGCCTGACACCGT


TTCATAACGGAAAAA CAGTTTGAGGAAGTG CCGCCTGACACCGTG


TCATAACGGAAAA.AT AGTTTGAGGAAGTGG CGCCTGACACCGTGG


CATAACGGAAAAATA GTTTGAGGAAGTGGC GCCTGACACCGTGGG


50ATAACGGAAAAATAA TTTGAGGAAGTGGCT CCTGACACCGTGGGT


TAACGGAAAAATAAT TTGAGGAAGTGGCTG CTGACACCGTGGGTC




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TGACACCGTGGGTCA TTATCTTTCACCTTT CCAAGGCTGTTACCA


GACACCGTGGGTCAT TATCTTTCACCTTTC CAAGGCTGTTACCAT


ACACCGTGGGTCATT ATCTTTCACCTTTCT AAGGCTGTTACCATT


CACCGTGGGTCATTA TCTTTCACCTTTCTA AGGCTGTTACCATTT


ACCGTGGGTCATTAC CTTTCACCTTTCTAG GGCTGTTACCATTTT


CCGTGGGTCATTACA TTTCACCTTTCTAGG GCTGTTACCATTTTA


CGTGGGTCATTACAA TTCACCTTTCTAGGG CTGTTACCATTTTAA


GTGGGTCATTACAAA TCACCTTTCTAGGGA TGTTACCATTTTAAC


TGGGTCATTACAAAA CACCTTTCTAGGGAC GTTACCATTTTAACG


10GGGTCATTACAAAAA ACCTTTCTAGGGACA TTACCATTTTAACGC


GGTCATTACAAAAAA CCTTTCTAGGGACAT TACCATTTTAACGCT


GTCATTACAAAAAAA CTTTCTAGGGACATG ACCATTTTAACGCTG


TCATTACAAAAAAAC TTTCTAGGGACATGA CCATTTTAACGCTGC


CATTACAAAAAAACA TTCTAGGGACATGAA CATTTTAACGCTGCC


15ATTACAAAAAAACAC TCTAGGGACATGAAA ATTTTAACGCTGCCT


TTACAA.AA.AA.ACACG CTAGGGACATGAAAT TTTTAACGCTGCCTA


TACAAAAAAACACGT TAGGGACATGAAATT TTTAACGCTGCCTAA


ACAA.AAAA.ACACGTG AGGGACATGAAATTT TTAACGCTGCCTAAT


CAAAAAAACACGTGG GGGACATGAAATTTA TAACGCTGCCTAATT


20AAAAAAACACGTGGA GGACATGAAATTTAC AACGCTGCCTAATTT


AAAAAACACGTGGAG GACATGAAATTTACA ACGCTGCCTAATTTT


AAAAACACGTGGAGA ACATGAAATTTACAA CGCTGCCTAATTTTG


AAAACACGTGGAGAT CATGAAATTTACAAA GCTGCCTAATTTTGC


AAACACGTGGAGATG ATGAAATTTACAAAG CTGCCTAATTTTGCC


25AACACGTGGAGATGG TGAAATTTACAAAGG TGCCTAATTTTGCCA


ACACGTGGAGATGGA GAAATTTACAAAGGG GCCTAATTTTGCCAA


CACGTGGAGATGGAA AAATTTACAAAGGGC CCTAATTTTGCCAAA


ACGTGGAGATGGAAA AATTTACAAAGGGCC CTAATTTTGCCAAAA


CGTGGAGATGGAAAT ATTTACAAAGGGCCA TAATTTTGCCAAAAT


30GTGGAGATGGAAATT TTTACAAAGGGCCAT AATTTTGCCAAAATC


TGGAGATGGAAATTT TTACAAAGGGCCATC ATTTTGCCAAAATCC


GGAGATGGAAATTTT TACAAAGGGCCATCG TTTTGCCAAAATCCT


GAGATGGAAATTTTT ACAAAGGGCCATCGT TTTGCCAAAATCCTG


AGATGGAAATTTTTA CAAAGGGCCATCGTT TTGCCAAAATCCTGA


35GATGGAAATTTTTAC AAAGGGCCATCGTTC TGCCAAAATCCTGAA


ATGGAAATTTTTACC AAGGGCCATCGTTCA GCCAAAATCCTGAAC


TGGAAATTTTTACCT AGGGCCATCGTTCAT CCAAAATCCTGAACT


GGAAATTTTTACCTT GGGCCATCGTTCATC CAAAATCCTGAACTT


GAAATTTTTACCTTT GGCCATCGTTCATCC AAAATCCTGAACTTT


40AAATTTTTACCTTTA GCCATCGTTCATCCA AAATCCTGAACTTTC


AATTTTTACCTTTAT CCATCGTTCATCCAA AATCCTGAACTTTCT


ATTTTTACCTTTATC CATCGTTCATCCAAG ATCCTGAACTTTCTC


TTTTTACCTTTATCT ATCGTTCATCCAAGG TCCTGAACTTTCTCC


TTTTACCTTTATCTT TCGTTCATCCAAGGC CCTGAACTTTCTCCC


45TTTACCTTTATCTTT CGTTCATCCAAGGCT CTGAACTTTCTCCCT


TTACCTTTATCTTTC GTTCATCCAAGGCTG TGAACTTTCTCCCTC


TACCTTTATCTTTCA TTCATCCAAGGCTGT GAACTTTCTCCCTCA


ACCTTTATCTTTCAC TCATCCAAGGCTGTT AACTTTCTCCCTCAT


CCTTTATCTTTCACC CATCCAAGGCTGTTA ACTTTCTCCCTCATC


50CTTTATCTTTCACCT ATCCAAGGCTGTTAC CTTTCTCCCTCATCG


TTTATCTTTCACCTT TCCAAGGCTGTTACC TTTCTCCCTCATCGG




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TTCTCCCTCATCGGC GCATGGCAGCTGGTT CCATCCGACTGCCCC


TCTCCCTCATCGGCC CATGGCAGCTGGTTG CATCCGACTGCCCCT


CTCCCTCATCGGCCC ATGGCAGCTGGTTGC ATCCGACTGCCCCTG


TCCCTCATCGGCCCG TGGCAGCTGGTTGCT TCCGACTGCCCCTGC


CCCTCATCGGCCCGG GGCAGCTGGTTGCTC CCGACTGCCCCTGCT


CCTCATCGGCCCGGC GCAGCTGGTTGCTCC CGACTGCCCCTGCTG


CTCATCGGCCCGGCG CAGCTGGTTGCTCCA GACTGCCCCTGCTGT


TCATCGGCCCGGCGC AGCTGGTTGCTCCAT ACTGCCCCTGCTGTG


CATCGGCCCGGCGCT GCTGGTTGCTCCATT CTGCCCCTGCTGTGC


10ATCGGCCCGGCGCTG CTGGTTGCTCCATTT TGCCCCTGCTGTGCT


TCGGCCCGGCGCTGA TGGTTGCTCCATTTG GCCCCTGCTGTGCTG


CGGCCCGGCGCTGAT GGTTGCTCCATTTGA CCCCTGCTGTGCTGC


GGCCCGGCGCTGATT GTTGCTCCATTTGAG CCCTGCTGTGCTGCT


GCCCGGCGCTGATTC TTGCTCCATTTGAGA CCTGCTGTGCTGCTC


15CCCGGCGCTGATTCC TGCTCCATTTGAGAG CTGCTGTGCTGCTCA


CCGGCGCTGATTCCT GCTCCATTTGAGAGA TGCTGTGCTGCTCAA


CGGCGCTGATTCCTC CTCCATTTGAGAGAC GCTGTGCTGCTCAAG


GGCGCTGATTCCTCG TCCATTTGAGAGACA CTGTGCTGCTCAAGG


GCGCTGATTCCTCGT CCATTTGAGAGACAC TGTGCTGCTCAAGGC


20CGCTGATTCCTCGTG CATTTGAGAGACACG GTGCTGCTCAAGGCC


GCTGATTCCTCGTGT ATTTGAGAGACACGC TGCTGCTCAAGGCCA


CTGATTCCTCGTGTC TTTGAGAGACACGCT GCTGCTCAAGGCCAC


TGATTCCTCGTGTCC TTGAGAGACACGCTG CTGCTCAAGGCCACA


GATTCCTCGTGTCCG TGAGAGACACGCTGG TGCTCAAGGCCACAG


25ATTCCTCGTGTCCGG GAGAGACACGCTGGC GCTCAAGGCCACAGG


TTCCTCGTGTCCGGA AGAGACACGCTGGCG CTCAAGGCCACAGGC


TCCTCGTGTCCGGAG GAGACACGCTGGCGA TCAAGGCCACAGGCA


CCTCGTGTCCGGAGG AGACACGCTGGCGAC CAAGGCCACAGGCAC


CTCGTGTCCGGAGGC GACACGCTGGCGACA AAGGCCACAGGCACA


30TCGTGTCCGGAGGCA ACACGCTGGCGACAC AGGCCACAGGCACAC


CGTGTCCGGAGGCAT CACGCTGGCGACACA GGCCACAGGCACACA


GTGTCCGGAGGCATG ACGCTGGCGACACAC GCCACAGGCACACAG


TGTCCGGAGGCATGG CGCTGGCGACACACT CCACAGGCACACAGG


GTCCGGAGGCATGGG GCTGGCGACACACTC CACAGGCACACAGGT


35TCCGGAGGCATGGGT CTGGCGACACACTCC ACAGGCACACAGGTC


CCGGAGGCATGGGTG TGGCGACACACTCCG CAGGCACACAGGTCT


CGGAGGCATGGGTGA GGCGACACACTCCGT AGGCACACAGGTCTC


GGAGGCATGGGTGAG GCGACACACTCCGTC GGCACACAGGTCTCA


GAGGCATGGGTGAGC CGACACACTCCGTCC GCACACAGGTCTCAT


40AGGCATGGGTGAGCA GACACACTCCGTCCA CACACAGGTCTCATT


GGCATGGGTGAGCAT ACACACTCCGTCCAT ACACAGGTCTCATTG


GCATGGGTGAGCATG CACACTCCGTCCATC CACAGGTCTCATTGC


CATGGGTGAGCATGG ACACTCCGTCCATCC ACAGGTCTCATTGCT


ATGGGTGAGCATGGC CACTCCGTCCATCCG CAGGTCTCATTGCTT


45TGGGTGAGCATGGCA ACTCCGTCCATCCGA AGGTCTCATTGCTTC


GGGTGAGCATGGCAG CTCCGTCCATCCGAC GGTCTCATTGCTTCT


GGTGAGCATGGCAGC TCCGTCCATCCGACT GTCTCATTGCTTCTG


GTGAGCATGGCAGCT CCGTCCATCCGACTG TCTCATTGCTTCTGA


TGAGCATGGCAGCTG CGTCCATCCGACTGC CTCATTGCTTCTGAC


50GAGCATGGCAGCTGG GTCCATCCGACTGCC TCATTGCTTCTGACT


AGCATGGCAGCTGGT TCCATCCGACTGCCC CATTGCTTCTGACTA




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ATTGCTTCTGACTAG CTCTCAGTGAAGGTG


TTGCTTCTGACTAGA TCTCAGTGAAGGTGG


TGCTTCTGACTAGAT CTCAGTGAAGGTGGG


GCTTCTGACTAGATT TCAGTGAAGGTGGGG


CTTCTGACTAGATTA CAGTGAAGGTGGGGA


TTCTGACTAGATTAT AGTGAAGGTGGGGAG


TCTGACTAGATTATT GTGAAGGTGGGGAGA


CTGACTAGATTATTA TGAAGGTGGGGAGAA


TGACTAGATTATTAT GAAGGTGGGGAGAAG


10GACTAGATTATTATT AAGGTGGGGAGAAGC


ACTAGATTATTATTT AGGTGGGGAGAAGCT


CTAGATTATTATTTG GGTGGGGAGAAGCTG


TAGATTATTATTTGG GTGGGGAGAAGCTGA


AGATTATTATTTGGG TGGGGAGAAGCTGAA


15GATTATTATTTGGGG GGGGAGAAGCTGAAC


ATTATTATTTGGGGG GGGAGAAGCTGAACC


TTATTATTTGGGGGA GGAGAAGCTGAACCG


TATTATTTGGGGGAA GAGAAGCTGAACCGG


ATTATTTGGGGGAAC AGAAGCTGAACCGGC


20TTATTTGGGGGAACT


TATTTGGGGGAACTG


ATTTGGGGGAACTGG


TTTGGGGGAACTGGA


TTGGGGGAACTGGAC


25TGGGGGAACTGGACA


GGGGGAACTGGACAC


GGGGAACTGGACACA


GGGAACTGGACACAA .


GGAACTGGACACAAT


30GAACTGGACACAATA


AACTGGACACAATAG


ACTGGACACAATAGG


CTGGACACAATAGGT


TGGACACAATAGGTC


35GGACACAATAGGTCT


GACACAATAGGTCTT


ACACAATAGGTCTTT


CACAATAGGTCTTTC


ACAATAGGTCTTTCT


40CAATAGGTCTTTCTC


AATAGGTCTTTCTCT


ATAGGTCTTTCTCTC


TAGGTCTTTCTCTCA


AGGTCTTTCTCTCAG


45GGTCTTTCTCTCAGT


GTCTTTCTCTCAGTG


TCTTTCTCTCAGTGA


CTTTCTCTCAGTGAA


TTTCTCTCAGTGAAG


50TTCTCTCAGTGAAGG


TCTCTCAGTGAAGGT




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EXAMPLE 9
Sub-confluent HaCaT cells were treated as described above with
phosphorothioate
oligonucleotides IGFR.AS (antisense: 5'-ATCTCTCCGCTTCCTTTC-3'; ( < 400 > 10);
ref
S 13) and IGFR.S (sense control: 5'-GAAAGGAAGCGGAGAGAT-3'; ( < 400 > 11); ref
13)
IGF-I binding to the cell monolayers was then measured as 'zsI_IGF-I.
EXAMPLE 10
The results of this experiment are shown in Figures 7 and 8.
HaCaT cells were initially plated in DMEM with 10 % v/v serum, then AS oligo
experiments
were performed in complete "Keratinocyte-SFM" (Gibco) to exclude the influence
of
exogenous IGFBPs. Oligos were synthesised as phosphorothioate (nuclease-
resistant)
derivatives (Bresatec, South Australia) and were as follows: antisense: AS2,
5'-
GCGCCCGCTGCATGACGCCTGCAAC-3' (IGFBP-3 start codon); controls: AS2NS, 5'-
CGGAGATGCCGCATGCCAGCGCAGG-3'; AS4,
5'-AGGCGGCTGACGGCACTA-3'; AS4NS, 5'-GACAGCGTCGGAGCGATC-3';
IGFRAS, 5'-ATCTCTCCGCTTCCTTTC-3';
IGFRS, 5'-GAAAGGAAGCGGAGAGAT-3'. Oligos to IGFBP-3 were based on the
published sequence of Spratt et al [12]. AS oligos were added to HaCaT
monolayers in O.SmI
medium in 24-well plates at the concentrations and addition frequencies
indicated. IGFBP-3
measured in cell-conditioned medium using a dot-blot assay, adapted from the
Western ligand
blot method of Hossenlopp et al [11], in which 100.1 of conditioned medium was
applied to
nitrocellulose filters with a vacuum dot-blot apparatus. After drying the
membranes at 37°C,
relative amounts of IGFBP are determined by 'zsI-IGF-I-binding,
autoradiography and
computerised imaging densitometry. Triplicate wells (except in Figure 7, where
duplicate
wells were measured as shown) were analysed and corrected for changes in cell
number per
well. Relative cell number per well was determined using an amido black dye
method,
developed specifically for cultured monolayers of HaCaT cells [14]. Cell
numbers differed
by less than 10% after treatment. For oligos to the IGF receptor, receptor
quantitation in


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intact HaCaT monolayers was by overnight incubation with 'ZSI-IGF-I
(30,OOOcpm/well) at
4°C.
EXAMPLE 11
Experiments involving ribozymes are generally conducted as described in
Internaitonal Patent
Application No. WO 89/05852 and in Haselhoff and Gerlach [8]. Ribozymes are
constructed
with a hybridising region which is complementary in nucleotide sequence to at
least part of
a target RNA which, in this case, encodes IGFBP-2. Activity of ribozymes is
measurable on,
for example, Northern blots or using animal models such as in the nude mouse
model (15; 16)
or the "flaky skin" mouse model (17; 18).
EXAMPLE 12
The methods described in Example 11 are used for the screening of ribozymes
which inhibit
IGFBP-3 production. The activity of the ribozymes is determined as in Example
11.
EXAMPLE 13
The methods described in Example 11 are used for the screening of ribozymes
which inhibit
IGF-1 production. The activity of the ribozymes is determined as in Example
11.
EXAMPLE 14
The methods described in Example 11 are used for the screening of ribozymes
which inhibit
IGF-1 production. The activity of the ribozymes is determined as in Example
11.
EXAMPLE 15
Twenty-one antisense oligonucleotides targeted to mRNA sequences enducing the
IGF-1
receptor, and four random oligonucleotides were synthesized. The antisense
oligonucleotides
are CS-propynyl-dU, dC l5mer phosphorothioate oligodeoxyribonucleotides. In
these
oligonucleotides, a phosphorothioate backbone replaces the phosphodiester
backbone of
naturally occurring DNA. The positions of the 21 sequence specific antisense
oligonucleotides relative to the IGF-1 receptor mRNA structure are shown in
Figure 9.


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EXAMPLE 16
Experiments were performed to determine the uptake of the antisense
oligonucleotides of
Example 15 into keratinocytes. Cells of the differentiated human keratinocyte
cell line,
HaCaT, were incubated for 24 hours in Dulbecco's Modified Eagle Medium (DMEM)
supplemented with 10 % (w/v) fetal calf serum (FCS) containing fluorescently
labelled
oligonucleotide (R451, a randomized sequence oligonucleotide, 30nM) and
cytofectin GSV
(2,ug/ml, Glen Research, 44901 Falcon Place, Sterling, VA 20166, Cat. No. 70-
3815-78).
Cells were then transferred to oligonucleotide-free medium and fluorescence
microcopy and
phase contrast images of the cells were obtained. Figure 10 shows fluorescence
microscopy
(Panel A) and phase contrast (Panel B) images of uptake of fluorescently
labelled
oligonucleotide in the majority of cells in a HaCaT monolayer. The degree of
uptake obtained
with the cationic lipid cytofectin was far greater than the uptake obtained
with the next best
lipid tried, Tfx-50.
A further experiment was performed to assess the uptake and toxicity
associated with the use
of cytofectin GSV over five days. Confluent HaCaT keratinocytes were incubated
in DMEM
containing fluorescently labelled oligonucleotide 8451 (30nM or 100 nM) plus
cytofectin
GSV (2,ug/ml or S,ug/ml) over 120 hours, viewed by fluorescence microscopy,
tryptan blue
stained, and counted. The graphs in Figure 11 depict uptake (Panel A) and
toxicity (Panel
B). The proportion of cells containing oligonucleotide remained high over the
120 hour
period. The combination of 30 nM oligonucleotide and 2,ug/ml GSV provided
optimal uptake
and minimal toxicity.
EXAMPLE 17
The twenty-one oligonucleotides of Example 15 were then screened for their
ability to inhibit
IGF-I receptor mRNA levels in HaCaT cells, in accordance with the teachings
herein. HaCaT
cells were grown to 90 % confluence in DMEM supplemented with 10 % (v/v) FCS.
Antisense oligonucleotides (30nM) were completed with cytofectin GSV (2,ug/ml)
and added
tot he cells in the presence of serum. HaCaT keratinocytes were treated with
the
oligonucleotide/GSV complexes or randomized sequence oligonucleotides (R451,
R766),


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liposome alone (GSV), or were left untreated (UT). Duplicate treatments were
performed.
Repeat additions of the oligonucleotides/GSV complex were performed at 24, 48
and 76 hours
following the first addition. Total RNA was isolated as per the RNAzoIB
protocol (Biotecx
Laboratories, Inc. 6023 South Loop East, Houston, TX 77033) 96 hours following
the first
S addition.
IGF-I receptor mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA
levels were simultaneously determined by a ribonuclease (RNase) protection
assay. The
RNase Protection Assay kit, in vitro transcription kit, and IGF-I receptor and
GAPDH DNA
templates were obtained from Ambion, Inc. (2130 Woodward St., Houston, TX
78744). The
amount of IGF-I receptor mRNA in any given sample was expressed as the amount
of IGF-I
receptor mRNA relative to the amount of GAPDH mRNA. Each oligonucleotide was
tested
in at least two separate experiments.
Figure 12 depicts representative results of the screening process. Panel A
shows an
electrophoretic analysis of IGF-I receptor and GAPDH mRNA fragments after
RNase
protection. Molecular weight markers are shown on the right hand side. The
full-length
probe is shown on the left hand side; G-probe indicates the IGF-I receptor
probe. GAPDH
protected fragments (G) are seen at 316 bases and IGF-I protected fragments
(I) are seen at
276 bases. Exhibit E, Panel B provides a graph indicating the relative level
of IGF-I receptor
mRNA following each treatment.
The results obtaining from the above screening assays are summarized in Figure
13. The
graph depicts the relative level of IGF-I receptor mRNA after treatment with
oligonucleotides
complementary to the human IGF-I receptor mRNA (26-86), four randomized
sequence
oligonucleotides (R1, R4, R7, R9), liposome alone (GSV), or no treatment (UT).
Asterisks
indicate a significant different in relative IGF-I receptor mRNA as compared
to GSV treated
cells (n=4-10, p < 0.05).


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As demonstrated in Figure 13, treatment with eighteen of the twenty-one
oligonucleotides
resulted in a significant different in levels of IGF-I receptor mRNA relative
to GSV treated
cells. Three of the antisense oligonucleotides tested in the screening assay
reduce IGF-I
receptor mRNA to less than 35 % of GSV-treated cells. These antisense
oligonucleotides have
the following sequences, presented in the 5' to 3' direction:
#27 UCCGGAGCCAGACUU
#64 CACAGUUGCUGCAAG
#78 UCUCCGCUUCCUUUC
As further demonstrated in Figure 13, six of the antisense oligonucleotides
tested in the
screening assay reduce IGF-I receptor mRNA to between 35 and 50% of GSV-
treated cells.
These antisense oligonucleotides have the following sequences, presented in
the 5' to 3'
direction:
#28 AGCCCCCACAGCGAG
#32 GCCUUGGAGAUGAGC
#40 UAACAGAGGUCAGCA
#42 GGAUCAGGGACCAGU
#46 CGGCAAGCUACACAG
#50 GGCAGGCAGGCACAC
EXAMPLE 19
Another experiment was performed demonstrating that antisense oligonucleotides
targeted to
genetic sequences encoding the IGFOI receptor and that reduce IGF-I receptor
mRNA levels
also inhibit the IGF-I receptor level on the surface of the treated cultured
keratinocytes.
HaCaT cells were grown to confluence in 24-well plates in DMEM containing 10 %
(v/v)
FCS. Oligodeoxynucleotide and cytofectin GSV were mixed together in serum-free
DMEM,
and incubated at room temperature for 10 minutes before being diluted ten-fold
in medium
and placed on the cells. Cells were incubated for 72 hours with 30nM random
sequence or


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antisense oligonucleotide and 2,um/ml GSV, or with GSV alone in DMEM
containing 10%
(v/v) FCS with solutions replaced every 24 hours. This was followed by
incubation with
oligonucleotide/GSV in serum-free DMEM for 48 hours. All incubations were
performed at
37°C. Cells were washed twice with lml cold PBS. Serum-free DMEM
containing 10-
'°M'ZSI-IGF-I was added with or without the IGF-I analogue, des (1-3)
IGF-I, at 10-"M to 10-
'M. Cells were incubated at 4°C for 17 hours with gentle shaking, then
washed three times
with lml cold PBS and lysed in 250~c1 O.SM NaOH/0.1 % (v/v) Triton X-100 at
room
temperature for 4 hours. Specific binding of the solubilised cell extract was
measured using
a gamma counter. As shown in Figure 14, treatment of HaCaT keratinocytes with
oligonucleotide reduced cell surface IGF-I receptor levels to 30 % of levels
in untreated
keratinocytes or in keratinocytes treated with liposome alone or a random
oligonucleotide,
8766. As shown in Figure 15, treatment with oligonucleotide #27 also
significantly reduced
cell surface IGF-I receptor levels relative to untreated keratinocytes or
treatment with
liposome alone or random nucleotide 8451. As demonstrated in Example 17,
oligonucleotides #64 and #27 reduce IGF-I receptor mRNA levels in cultured
keratinocytes
to less than 35% of GSV-treated cells. Accordingly, the ability of an
oligonucleotide to
reduce IGF-I receptor mRNA levels in correlated with its ability to reduce
cell surface IGF-I
receptor levels.
The forgoing Examples demonstrate that antisense oligonucleotides targeted to
the IGF-I
receptor can be delivered to human keratinocytes in vitro, can inhibit IGF-I
receptor mRNA
levels in human keratinocytes in vitro, and that inhibition of mRNA levels is
correlated with
reduction of cell surface IGF-I receptor levels.
EXAMPLE 19
Further experiments demonstrated the efficacy of antisense oligonucleotides
targeted tot he
IGF-I receptor in an in vivo model of psoriasis. An animal model of psoriasis
is the human
psoriatic skin xenograft model. The skin used in this model contains the true
disease state.
In this model, reduction in epidermal thickness of psoriatic grafts in
response to treatment is
positively correlated with efficacy of treatment. Both normal and psoriatic
human skin were


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grated into a thymic (nude) mice in accordance with a thymic (nude) mice in
accordance with
the methods of Baker et al (1992) Brit. J. Dermatol. 126:105 and Nanney et al
(1992) J.
Invest. Dermatol, 92:296. Successful grafting was achieved, as demonstrated in
Figure 16,
which shows hemotoxylin and eosin (H&E) stained sections of a 49-day old
psoriatic human
skin graft (Panel B) compared to the histology of the skin graft prior to
grafting (Panel A).
The histological features of psoriasis present in the pregraft section (e.g.,
parakeratosis,
acanthosis and pronounced rete ridges) are present in the grafts more than
seven weeks post
grafting.
Using the model, oligonucleotide uptake was measured in epidermal
keratinocytes in vivo
after intradermal injection. Fluorescently labelled oligonucleotide (R451,
SO,uI, lO,uM
injection) was intradermally injected into psoriatic and normal skin grafts on
a thymic mice.
Live confocal microscopy and fluorescence microscopy of fixed sections was
then employed.
Using both techniques, oligonucleotide was found to localize in the nucleus of
over 90 % of
basal keratinocytes. Figure 17 shows the nuclear localization of
oligonucleotide in psoriatic
skin cells using conventional fluorescence microscopy of a graft that was
removed and
sectioned after 24 hours.
After establishing oligonucleotide uptake in the in vivo model, a small number
of pilots
experiments were performed to determine a schedule for treatment of grated
mice with
antisense oligonucleotides targeted to genetic sequences encoding the IGF-I
receptor. The
treatment schedule was finalized as follows:


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Graft Number Treatment Volume ODN Duration
of Concentrationof
Injection Treatment


1-3 Vehicle (PBS) SO,uI - 20 days


4-6 RandomODN#R451 SO,uI lO,uM 20 days


S 7-9 ODN#27 SO,uI 10~M 20 days


10-12 ODN#74 501 lO,uM 20 days


13-15 ODN#50 SO,uI 10~M 20 days


As determined above, oligonucleotide #27 (ODN #27) reduced IGF-I receptor mRNA
in
vitro to less than 35 % of GSV-treated cells. Oligonucleotide #5O (ODN#50)
reduced IGF-
I receptor mRNA in vitro to between 35 and 50% of GSV-treated cells.
Oligonucleotide
#74 (ODN #74) was not inhibitory to IGF-I receptor mRNA in vitro. In the in
vivo
model, each mouse received two grafts. Random oligonucleotide or vehicle was
injected
intradermally in one graft and acted as a control. The second graft was
injected with the
targeted oligonucleotide. Each graft received an injection every second day
for the
duration of the treatment.
Histology of representative grafts from each treatment type are shown in
Figures 18(a)-(d)
and 19(a) - (d). Each sheet shows three images of H&E stained sections: the
pregraft
histology, the control treated graft, and the targeted oligonucleotide treated
graft. Figures
18(a)-(d) are shown at 100x magnification; figures 19(a)-(d) are shown at 400x
magnification. The total cross sectional area of epidermis of each graft was
assessed using
MCID analysis software. The pooled results from all of the treated grafts are
shown in
Figure 20.
As shown in Figures 18(a)-(d) and 19(a)-(d), the vehicle-treated (control)
grafts were
marginally thinner than thepregraft sections. The degree of regression in
these
experiments (ie., less than 10 % ) is not significant. A similar amount of
marginal thinning


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of epidermis compared to pregraft also occurred in pilot experiments in which
psoriatic
grafts were not injected, and thsu it is unlikely that the vehicle itself has
any effect.
Histological features of psoriasis present in skin samples prior to grafting
(clubbing of rete
ridges, parakeratosis, acanthosis) were present in these grafts.
The random oliognucleotide treated grafts varied in epidermal thickness after
20 days of
treatment. Grafts were either a similar thickness to the pregraft histology,
or marginally
thinner. Random oligonucleotide treated grafts were in each case significantly
thicker than
their targeted oligonucleotide treated pairs.
As shown in Figure 20, the targeted oligonucleotide treated grafts were
significantly
thinner than the pregraft sections and showed less parakeratosis and clubbing
of rete
ridges. Antisense oligonucleotides which were effective at reducing IGF-I
receptor
mRNA levels in vitro (#27 and #50) produced greatere epidermal thinning than
an
oligonucleotide which was not inhibitory to IGF-I receptor mRNA in vitro
(#74).
Accordingly, there is a direct correlation between the ability of an
oligonucleotide targeted
to the IGF-I receptor to inhibit IGF-I receptor mRNA levels in vitro and the
efficacy of the
oligonucleotide as an anti-psoriasis agent in an in vivo model.
EXAMPLE 20
Another experiment demonstrated that treatment of psoriatic grafts with an
oligonucleotide
targeted to a genetic sequence encoding the IGF-I receptor results in
inhibition of
proliferation. Pregrafts from psoriatic patients, control grafts treated with
84541, and
grafts treated with oligonucleotide #27 were obtained as described in Example
19. An
antibody to the cell cycle-specific nuclear antigen Ki67 was used to
immunohistochemically detect actively dividing cells and tereby assess
proliferation. The
aKi67 antibody (DAKO, Glostrup, Denmark) recognizes the Ki67 antigen
transiently
expressed in nuclei of proliferating cells during late G,, S, M and GZ phases
of the cycle
and thsu provides a marker for proliferation. Pregraft and graft sections were
immunohistochemically processed by standard methods using aKi67 (according to
the


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manufacturer's instructions), peroxidase-conjugated anti-rabbit second stage
antibody, and
a chromogenic peroxidase substrate.
The results of this experiment are presented in Figure 21 as
immunohistochemical sections
at 100x magnification. The top panel of Figure 21 depicts a pregraft section
obtained from
a psoriatic patient. The epidermis is thicker than normal and nucleic are
evident in the
stratum corneum. Ki67 positive cells, appearing as brown dots, are evidence in
the basal
and suprabasal layers, and indicate actively proliferating cells. The control
(R450-treated)
graft in the bottom panel of Figure 21 also exhibits evidence of
proliferation, including
parakeratosis and Ki67-positive cells appearing as brown-staining nuclei. The
center panel
of Figure 21 exhibits the oligonucleotide #27-treated graft. This graft
exhibits
significantly reduced proliferation as evidenced by normal (thin) epidermis,
lack of
invaginations, and substantial loss of Ki67-positive cells.
These results indicate that treatment of human psoriatic grafts with an
oligonucleotide
targeted to mRNA encoding the IGF-I receptor results in inhibition of
epidermal
proliferation.
EXAMPLE 21
Topical formulations of complexes of oligonucleotides with cytofectin GSV in
aqueous or
methylcellulose gel formulations were prepared and assessed foruptake of the
oligonucleotide by keratinocytes in vivo. The topical formulations contained
oligonucleotides complexed with cytofectin GSV in an aqueous solution or
methylcellulose
carrier, as taught herein. With both aqueous and methylcellulose gel
formulations,
locatlization of oligonucleotide 8451 to nuclei and cytoplasm of keratinocytes
in normal
human skin grafts on nuce mice was observed. Figure 22 shows an image from
confocal
microscopy demonstrating oligonucleotide locatlization in the nuclei and
cytoplasm of
keratinocytes in normal human skin grafts after topical application of
fluroescently labeled
oligonucleotide (lO~cM 8451) complexed with cytofectin GSV (l0,ug/ml). Figure
23
shows an image from confocal microscopy demonstrating that topical application
of the


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same oligonucleotide/GSV concentrations in a 3 % (w/v) methylcellulose gel
produced
similar uptake in the target keratinocyte population. Using an aqueous
formulation of
oligonucleotide/GSV complexes, penetration of oligonucleotide into the viable
epidermis
was observed, whereas application of formulations of oliognucleotide complexed
with
other cationic lipids resulted in localization of oligonucleotide in the
stratum corneum.
EXAMPLE 22
Thirteen antisense oligonucleotides targeted to IGFBP-3 were synthesized. The
antisense
oligonucleotides are CS-propynyl-dU, DclS mer phosphorothioate
oligodeoxyribonucleotides. Figure 24 attached hereto is a schematic diagram
indicating
the position of the thirteen oligonucleotides relative to the IGFBP-3 mRNA
structure.
These oligonucleotides were screened for their ability to inhibit IGFBP-3 mRNA
levels of
HaCaT cells in accordance with the teachings herein. HaCaT cells were grown to
90
confluence in DMEM supplemented with 10% (v/v) FCS, then placed in complete
keratinocyte serum free medium (KSFM, Gibco), which has a defined amount of
EGF, for
24 hours. Oligonucleotides (30nM or 100nM) were complexed with GSV cytofectin
(2,ug/ml) and added to cells in complete KSFM to allow oligonucleotides to
enter the
nucleus before removal of EGF. Repeat additions were performed at three hours
(in
serum free DMEM, which releases the EGF inhibition of IGFBP-3 mRNA) and again
after
another 24 hours. HaCaT cells were also treated with randomized sequence
oligonucleotides (R121, 8451, 8766 and R961), liposome alone (GSV) or were
left
untreated (UT). Total RNA was isolated as described in Example 17, 24 hours
after the
last treatment. Total RNA (l5,ug) was analyzed by Northern analysis and
phosphoroimager quantitation for IGFBP-3 and GADPH mRNA. IGFBP-3 mRNA is
expressed as the amount of IGFBP-3 mRNA relative to the amount of GAPDH mRNA.
Figures 25(a)-(d) provide graphs which depict results in this screening
process. In these
graphs, R1 and R12 refer to 8121; R4, R4(0) and R45 rfer to 8451; R7, R7(0)
and R76
refer to 8766; and R9 and R96 refer to 8961. The values were standardized to
GSV-


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treated cells, and data was pooled and statistically analyzed by ANOVA
followed by
Domet's test to compare each treatment to GSV-treated cells. The pooled data
are
presented as a bar graph in Figure 26. As demonstrated, at a concentration of
30nM,
treatment of HaCaT cells with 8 of the 12 targeted oligonucleotides tested
resulted in a
statistically significant reduction in levels of IGFBP-3 mRNA relative to GSV-
treated
cells. At a concentration of 100nM, treatment with 9 fo the 13 targeted
oligonucleotides
tested resulted in a statistically significant reduction in levels of IGFBP-3
mRNA relative
to GSV-treated cells.
These experiments demonstrate that antisense oligonucleotides targeted to
genetic
sequences encoding IGFBP-3 can inhibit IGFBP-3 mRNA levels in human
keratinocytes in
vitro.
EXAMPLE 23
IGF-I receptor is a potent mitotic signalling molecule for keratinocytes and
the human
receptor elicits separate intracellular signals that prevent apoptosis (19).
It is proposed in
accordance with the present invention that inactivation of IGF-I receptors in
epidermal
keratinocytes will achieve three important outcomes in subsequent UV treatment
of
lesions:
(i) Acute epidermal hyperplasia following UV has been suggested to increase
the risk
of keratinocyte carcinogenic transformation (22). By reducing IGF-I receptor
expression in the epidermis, the incidence of epidermal hyperplasia following
UV
exposure is likely to be reduced leading to an overall acceleration in
normalization
of the lesion and reduced carcinogenic risk.
(ii) Inhibition of anti-apoptotic action of IGF-I receptor will enhance the
reversal of
epidermal thickening and accelerate normalization of differentiation. Topical
or
injected IGF-I receptor antisense as adjunctive treatment will increase
apoptosis in


CA 02376284 2001-12-05
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the epidermal layer thereby enhancing the reduction in acanthosis observed in
UV
treatments.
(iii) Survival of keratinocytes, ie. those which evade apoptosis is likely to
occur when
cells have damaged DNA. Such mutations may be in the tumor suppressor region.
Consequently, the use of antisense therapy will result in less frequent
selection of
mutated keratinocytes and therefore reduced incidence of basal cell carcinomas
and
squamous.
Accordingly, antisense therapy, especially against IGF-I-receptor is useful in
combination
with UV therapy in the treatment of epidermal hyperplasia.
EXAMPLE 24
HaCaT cells were treated with antisense oligonucleotides directed to IGF-I
receptor
mRNA. Levels of IGF-I receptor mRNA were then monitored. In essence, confluent
HaCaT cells were treated every 24 hours for four days with 2 ,ug/ml GSV lipid
alone
(GSV) or complexed with 30 nM IGF-I receptor specific oligonucleotides (#26 to
#86) or
random sequence oligonucleotides (R121, 8451 and R76c~. Figure 27(a) is a
photographic
representation showing representative RNase protection assay gel showing IGF-I
receptor
(IGFR) and GAPDH mRNA in untreated or treated HaCaT cells. Figure 27(b) is a
densitometric quantification of IGF-I receptor mRNA in a HaCaT cells following
treatment with IGF-I receptor specific oligonucleotides (solid black) random
sequence
oligonucleotides (horizontal striped bar) or GSV alone (shaded bar) compared
to untreated
cells (UT, vertical striped bar).
EXAMPLE 25
In this example, reduction in total cellular IGF-I receptor protein was
monitored following
antisense oligonucleotide treatment. Confluence HaCaT cells were treated with
24 hours
for 4 days with 2 ,ug/ml GSV lipid alone (GSV) or complexed with 30 nM IGF-I
receptor
specific AONS (#27, #50 and #64) or the random sequence oligonucleotide, 8451.
Total


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cellular protein was isolated and analysed for IGF-I receptor by SDS PAGE
followed by
western blotting with antibody specific for the human IGF-I receptor. Figure
28(a) shows
duplicate treated cellular extracts following the IGF-I receptor at the
predicted size of 110
kD. Figure 28(b) is a densitometric quantification of IGF-I receptor protein.
EXAMPLE 26
The reduction in IGF-I receptor numbers was determined on the keratinocyte
cell surface
after antisense oligonucleotide treatment. HaCaT cells were tranfected with
IGF-I receptor
specific AONs #27, #50, #64, a random sequence oligonucleotides (R451) or
following
treatment with GSV a lipid alone every 24 hours for 4 days. Competition
binding assays
using 'ZSI-IGF-I and the receptor-specific analogue, des(1-3)IGF-I were
performed.
Results are shown in Figure 29.
EXAMPLE 27
In this example, the apoptotic protecting effects of IGF-I receptor on
keratinocyte cells
was tested by following the reduction in keratino cell numbers following
antisense
oligonucleotide treatment. HaCaT cells, initially at 40 % confluence, were
transfected with
the IGF-I receptor specific AON #64, control sequences 8451 and 6414 or
treated with
GSV a lipid alone every 24 hours for 2 days. The cell number was measured in
culture
wells using a dye binding assay. The results are presented in Figure 30. The
results clearly
confirm that the IGF-I receptor exhibits an anti-apoptotic effect. By reducing
IGF-I
receptor levels using antisense oligonucleotide treatment, the anti-apoptotic
effect is
interrupted and apoptosis results in the reduction in keratinocyte cell
number. Results are
shown in Figure 30.
EXAMPLE 28
This example shows a reversal of epidermal hyperplasia in psoriatic human skin
grafts on
nude mice following intradermal injection with antisense oligonucleotides.
Grafted
psoriasis lesions were injected with IGF-I receptor specific AONs, a random
sequence
oligonucleotide in PBS, or with PBS alone, every 2 days for 20 days, then
analysed


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histologically. The results are shown in Figure 31. In Figure 31(a), donor A
graft treated
with AON #50 showing epidermal thinning compared with the pregraft and control
(PBS)
treated graft and donor graft treated with AON #27 showing epidermal thinning
compared
with pregraft and control (R451) treated graft. In Figure 31(b), the mean
epidermal cross
sectional area over the full width of grafts is shown as determined by digital
image
analysis. The results show that epidermal hyperplasia is reversed following
the intradermal
injection of antisense oligonucleotides.
EXAMPLE 29
Figure 32 shows the reversal of epidermal hyperplasia correlating with reduced
IGF-I
receptor mRNA in grafted psoriasis lesions treated with antisense
oligonucleotides. Figure
32(a) shows a psoriasis lesion prior to grafting and after grafting and
treatment with IGF-I
receptor specific oligonucleotide #27 (AON #27) or random sequence (R451)
immunostained with antibodies to Ki67 to identify proliferating cells.
Proliferating cells
1 S are indicated by a dark brown nucleus (arrows). Figure 32(b) shows the
same lesion prior
to grafting and after oligonucleotide treatment as in Figure 32(a) but
subjected to in situ
hybridisation with 35S-labelled cRNA probe complementary to the human IGF-I
receptor
mRNA. The presence of IGF-I receptor mRNA is indicated by silver grains which
are
almost eliminated in the epidermis of the lesion treated with IGF-I receptor
specific
oligonucleotide # 27 (AON #27). This experiment shows that reversal of
epidermal
hyperplasia correlates with reduced IGF-I receptor mRNA in grafted psoriasis
lesions
treated with antisense oligonucleotides.
EXAMPLE 30
Figure 33 treatment with oligonucleotides. HaCaT cell monolayers were grown to
90%
confluence in DMEM containing 10 % fetal calf serum treated every 24 hours for
two days
with 2 ,ug/ml GSV lipid alone (GSV) or complexed with 30 nM oligonucleotide.
Total
RNA was isolated and analysed for IGF-I receptor and GAPDH mRNA using a
commercially available ribonuclease protection assay kit. The results show a
reduction in
IGF-I receptor mRNA in the HaCaT keratinocyte cells.


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EXAMPLE 31
Figure 34 treatment with oligonucleotides. HaCaT cell monolayers were grown to
90%
confluence in DMEM containing 10 % fetal calf serum treated every 24 hours for
4 days
with 2 ,ug/ml GSV lipid alone (GSV) or complexed with 30 nM oligonucleotide.
Cells
were lysed in a buffer containing 50 mM HEPES, 150 mM NaCI, 10 % v/v glycerol,
1 v/v
Trison X-100 and 100 ,ug/ml aprotinin on ice for 30 minutes, then 30 ,ug of
lysate was
loaded onto a denaturing 7 % w/v polyacrylamide gel followed by transfer onto
an
Immobilon-P membrane. Membranes were then incubated with anti-IGF-I receptor
antibodies C20 (available from Santa Cruz Biotechnology Inc., Santa Cruz,
California) for
1 hour at room temperature and developed using the Vistra ECF western blotting
kit
(Amersham). The results shown in Figure 34 confirm that IGF-I receptor protein
is
reduced in HaCaT keratinocytes following treatment with oligonucleotides.
EXAMPLE 32
This example shows a reduction in HaCaT keratinocyte cell number following
treatment
with oligonucleotides. The results are shown in Figure 35. HaCaT cell
monolayers were
grown at 40 % confluence in DMEM containing 10 % fetal calf serum treated
every 24
hours for 3 days with 2 ~g/ml GSV lipid alone (GSV) or complexed with 15 nM
oligonucleotide. Cell numbers were then measured every 24 hours using the
amido black
dye binding assay [32]. Results show that HaCaT keratino cells decrease in
number
following treatment with oligonucleotides due to a reduction in the anti-
apoptotic effect of
the IGF-I receptor.
Those skilled in the art will appreciate that the invention described herein
is susceptible to
variations and modifications other than those specifically described. It is to
be understood
that the invention includes all such variations and modifications. The
invention also
includes all of the steps, features, compositions and compounds referred to or
indicated in
this specification, individually or collectively, and any and all combinations
of any two or
more of said steps or features.


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REFERENCES:
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2. Rechler MM and Brown AL Growth Regulation 2:55-68, 1992.
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7. Ts'O POP, Aurelian L, Chang E and Miller PS. Nonionic oligonucleotide
analogs
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"Antisense
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and Denhardt DT, eds.), 1993.
8. Haseloff J and Gerlach L Nature 334:586-591, 1988.
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11. Hossenlopp P, Seurin D, Segovia-Quinson B, Hardouin S, Binoux M. Anal
Biochem
154:138-143, 1986.


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12. Spratt SK, Tatsuno GP, Yamanaka MK, Ark BC, Detmer J, Mascarenhas D, Flynn
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Talkington-Verser C, Spencer EM. Growth Factors 3:63-72, 1990.
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15. Baker BS, Brent L, Valdimarsson H, Powles AV, Al-Imam L, Walker M and Fry
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16. Nanney LB et al J. Invest. Bermatol 98:296-301, 1992.
17. Sundberg JP et al Immunol. Investigations 22:389-401, 1993.
18. Sundberg JP et al J. Invest. Dermatol 102:781-788, 1994.
19. O'Connor et al Mol Cell Biol 17:427-435, 1997.
20. Kuhn et al Int J Cancer 80:431-438, 1999.
21. Resnicoff et al Cancer Res 55:3739-3741, 1995.
22. Ouhtit et al Am JPathol 156:201-207, 2000.
23. Froehler et al Tetrahedrin Lett 34:1003-1006, 1992.
24. Gennaro (Ed) Remington's Pharmaceutical Sciences 18th Edition Mack
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25. Flanagan et al Nat Biotechnol 14:1139-1145, 1996.
26. Flanagan et al Nucleic Acids Res 24:2936-2941, 1996.
27. Flanagan et al Mol Cell Biochem 172:213-225, 1997.
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32. Schultz et al Jlmmunol Meth 167:1-13, 1994.


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SEQUENCE LISTING
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CA 02376284 2001-12-05
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ccttattttg cacaaaagac tgccaaggac atgaccagca gctggctaca gcctcgattt 1080
atatttctgt ttgtggtgaa ctgatttttt ttaaaccaaa gtttagaaag aggtttttga 1140
aatgcctatg gtttctttga atggtaaact tgagcatctt ttcactttcc agtagtcagc 1200
aaagagcagt ttgaattttc ttgtcgcttc ctatcaaaat attcagagac tcgagcacag 1260
cacccagact tcatgcgccc gtggaatgct caccacatgt tggtcgaagc ggccgaccac 1320
tgactttgtg acttaggcgg ctgtgttgcc tatgtagaga acacgcttca cccccactcc 1380
ccgtacagtg cgcacaggct ttatcgagaa taggaaaacc tttaaacccc ggtcatccgg 1440


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-3-
acatcccaac gcatgctcct ggagctcaca gccttctgtg gtgtcatttc tgaaacaagg 1500
gcgtggatcc ctcaaccaag aagaatgttt atgtcttcaa gtgacctgta ctgcttgggg 1560
actattggag aaaataaggt ggagtcctac ttgtttaaaa aatatgtatc taagaatgtt 1620
ctagggcact ctgggaacct ataaaggcag gtatttcggg ccctcctctt caggaatctt 1680
cctgaagaca tggcccagtc gaaggcccag gatggctttt gctgcggccc cgtggggtag 1740
gagggacaga gagacgggag agtcagcctc cacattcaga ggcatcacaa gtaatggcac 1800
aattcttcgg atgactgcag aaaatagtgt tttgtagttc aacaactcaa gacgaagctt 1860
atttctgagg ataagctctt taaaggcaaa gctttatttt catctctcat cttttgtcct 1920
ccttagcaca atgtaaaaaa gaatagtaat atcagaacag gaaggaggaa tggcttgctg 1980
gggagcccat ccaggacact gggagcacat agagattcac ccatgtttgt tgaacttaga 2040
gtcattctca tgcttttctt tataattcac acatatatgc agagaagata tgttcttgtt 2100
aacattgtat acaacatagc cccaaatata gtaagatcta tactagataa tcctagatga 2160
aatgttagag atgctatatg atacaactgt ggccatgact gaggaaagga gctcacgccc 2220
agagactggg ctgctctccc ggaggccaaa cccaagaagg tctggcaaag tcaggctcag 2280
ggagactctg ccctgctgca gacctcggtg tggacacacg ctgcatagag ctctccttga 2340
aaacagaggg gtctcaagac attctgccta cctattagct tttctttatt tttttaactt 2400
tttgggggga aaagtatttt tgagaagttt gtcttgcaat gtatttataa atagtaaata 2460
aagtttttac catt 2474
<210> 3
<211> 4989
<212> DNA
<213> synthetic construct
<400> 3
tttttttttt ttttgagaaa gggaatttca tcccaaataa aaggaatgaa gtctggctcc 60
ggaggagggt ccccgacctc gctgtggggg ctcctgtttc tctccgccgc gctctcgctc 120
tggccgacga gtggagaaat ctgcgggcca ggcatcgaca tccgcaacga ctatcagcag 180
ctgaagcgcc tggagaactg cacggtgatc gagggctacc tccacatcct gctcatctcc 240
aaggccgagg actaccgcag ctaccgcttc cccaagctca cggtcattac cgagtacttg 300
ctgctgttcc gagtggctgg cctcgagagc ctcggagacc tcttccccaa cctcacggtc 360
atccgcggct ggaaactctt ctacaactac gccctggtca tcttcgagat gaccaatctc 420
aaggatattg ggctttacaa cctgaggaac attactcggg gggccatcag gattgagaaa 480
aatgctgacc tctgttacct ctccactgtg gactggtccc tgatcctgga tgcggtgtcc 540
aataactaca ttgtggggaa taagccccca aaggaatgtg gggacctgtg tccagggacc 600
atggaggaga agccgatgtg tgagaagacc accatcaaca atgagtacaa ctaccgctgc 660
tggaccacaa accgctgcca gaaaatgtgc ccaagcacgt gtgggaagcg ggcgtgcacc 720
gagaacaatg agtgctgcca ccccgagtgc ctgggcagct gcagcgcgcc tgacaacgac 780
acggcctgtg tagcttgccg ccactactac tatgccggtg tctgtgtgcc tgcctgcccg 840
cccaacacct acaggtttga gggctggcgc tgtgtggacc gtgacttctg cgccaacatc 900


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-4-
ctcagcgccg agagcagcga ctccgagggg tttgtgatcc acgacggcga gtgcatgcag 960
gagtgcccct cgggcttcat ccgcaacggc agccagagca tgtactgcat cccttgtgaa 1020
ggtccttgcc cgaaggtctg tgaggaagaa aagaaaacaa agaccattga ttctgttact 1080
tctgctcaga tgctccaagg atgcaccatc ttcaagggca atttgctcat taacatccga 1140
cgggggaata acattgcttc agagctggag aacttcatgg ggctcatcga ggtggtgacg 1200
ggctacgtga agatccgcca ttctcatgcc ttggtctcct tgtccttcct aaaaaacctt 1260
cgcctcatcc taggagagga gcagctagaa gggaattact ccttctacgt cctcgacaac 1320
cagaacttgc agcaactgtg ggactgggac caccgcaacc tgaccatcaa agcagggaaa 1380
atgtactttg ctttcaatcc caaattatgt gtttccgaaa tttaccgcat ggaggaagtg 1440
acggggacta aagggcgcca aagcaaaggg gacataaaca ccaggaacaa cggggagaga 1500
gcctcctgtg aaagtgacgt cctgcatttc acctccacca ccacgtcgaa gaatcgcatc 1560
atcataacct ggcaccggta ccggccccct gactacaggg atctcatcag cttcaccgtt 1620
tactacaagg aagcaccctt taagaatgtc acagagtatg atgggcagga tgcctgcggc 1680
tccaacagct ggaacatggt ggacgtggac ctcccgccca acaaggacgt ggagcccggc 1740
atcttactac atgggctgaa gccctggact cagtacgccg tttacgtcaa ggctgtgacc 1800
ctcaccatgg tggagaacga ccatatccgt ggggccaaga gtgagatctt gtacattcgc 1860
accaatgctt cagttccttc cattcccttg gacgttcttt cagcatcgaa ctcctcttct 1920
cagttaatcg tgaagtggaa ccctccctct ctgcccaacg gcaacctgag ttactacatt 1980
gtgcgctggc agcggcagcc tcaggacggc tacctttacc ggcacaatta ctgctccaaa 2040
gacaaaatcc ccatcaggaa gtatgccgac ggcaccatcg acattgagga ggtcacagag 2100
aaccccaaga ctgaggtgtg tggtggggag aaagggcctt gctgcgcctg ccccaaaact 2160
gaagccgaga agcaggccga gaaggaggag gctgaatacc gcaaagtctt tgagaatttc 2220
ctgcacaact ccatcttcgt gcccagacct gaaaggaagc ggagagatgt catgcaagtg 2280
gccaacacca ccatgtccag ccgaagcagg aacaccacgg ccgcagacac ctacaacatc 2340
accgacccgg aagagctgga gacagagtac cctttctttg agagcagagt ggataacaag 2400
gagagaactg tcatttctaa ccttcggcct ttcacattgt accgcatcga tatccacagc 2460
tgcaaccacg aggctgagaa gctgggctgc agcgcctcca acttcgtctt tgcaaggact 2520
atgcccgcag aaggagcaga tgacattcct gggccagtga cctgggagcc aaggcctgaa 2580
aactccatct ttttaaagtg gccggaacct gagaatccca atggattgat tctaatgtat 2640
gaaataaaat acggatcaca agttgaggat cagcgagaat gtgtgtccag acaggaatac 2700
aggaagtatg gaggggccaa gctaaaccgg ctaaacccgg ggaactacac agcccggatt 2760
caggccacat ctctctctgg gaatgggtcg tggacagatc ctgtgttctt ctatgtccag 2820
gccaaaacag gatatgaaaa cttcatccat ctgatcatcg ctctgcccgt cgctgtcctg 2880
ttgatcgtgg gagggttggt gattatgctg tacgtcttcc atagaaagag aaataacagc 2940
aggctgggga atggagtgct gtatgcctct gtgaacccgg agtacttcag cgctgctgat 3000
gtgtacgttc ctgatgagtg ggaggtggct cgggagaaga tcaccatgag ccgggaactt 3060
gggcaggggt cgtttgggat ggtctatgaa ggagttgcca agggtgtggt gaaagatgaa 3120
cctgaaacca gagtggccat taaaacagtg aacgaggccg caagcatgcg tgagaggatt 3180
gagtttctca acgaagcttc tgtgatgaag gagttcaatt gtcaccatgt ggtgcgattg 3240
ctgggtgtgg tgtcccaagg ccagccaaca ctggtcatca tggaactgat gacacggggc 3300
gatctcaaaa gttatctccg gtctctgagg ccagaaatgg agaataatcc agtcctagca 3360


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-5-
cctccaagcc tgagcaagat gattcagatg gccggagaga ttgcagacgg catggcatac 3420
ctcaacgcca ataagttcgt ccacagagac cttgctgccc ggaattgcat ggtagccgaa 3480
gatttcacag tcaaaatcgg agattttggt atgacgcgag atatctatga gacagactat 3540
taccggaaag gaggcaaagg gctgctgccc gtgcgctgga tgtctcctga gtccctcaag 3600
gatggagtct tcaccactta ctcggacgtc tggtccttcg gggtcgtcct ctgggagatc 3660
gccacactgg ccgagcagcc ctaccagggc ttgtccaacg agcaagtcct tcgcttcgtc 3720
atggagggcg gccttctgga caagccagac aactgtcctg acatgctgtt tgaactgatg 3780
cgcatgtgct ggcagtataa ccccaagatg aggccttcct tcctggagat catcagcagc 3840
atcaaagagg agatggagcc tggcttccgg gaggtctcct tctactacag cgaggagaac 3900
aagctgcccg agccggagga gctggacctg gagccagaga acatggagag cgtccccctg 3960
gacccctcgg cctcctcgtc ctccctgcca ctgcccgaca gacactcagg acacaaggcc 4020
gagaacggcc ccggccctgg ggtgctggtc ctccgcgcca gcttcgacga gagacagcct 4080
tacgcccaca tgaacggggg ccgcaagaac gagcgggcct tgccgctgcc ccagtcttcg 4140
acctgctgat ccttggatcc tgaatctgtg caaacagtaa cgtgtgcgca cgcgcagcgg 4200
ggtggggggg gagagagagt tttaacaatc cattcacaag cctcctgtac ctcagtggat 4260
cttcagttct gcccttgctg cccgcgggag acagcttctc tgcagtaaaa cacatttggg 4320
atgttccttt tttcaatatg caagcagctt tttattccct gcccaaaccc ttaactgaca 4380
tgggccttta agaaccttaa tgacaacact taatagcaac agagcacttg agaaccagtc 4440
tcctcactct gtccctgtcc ttccctgttc tccctttctc tctcctctct gcttcataac 4500
ggaaaaataa ttgccacaag tccagctggg aagccctttt tatcagtttg aggaagtggc 4560
tgtccctgtg gccccatcca accactgtac acacccgcct gacaccgtgg gtcattacaa 4620
aaaaacacgt ggagatggaa atttttacct ttatctttca cctttctagg gacatgaaat 4680
ttacaaaggg ccatcgttca tccaaggctg ttaccatttt aacgctgcct aattttgcca 4740
aaatcctgaa ctttctccct catcggcccg gcgctgattc ctcgtgtccg gaggcatggg 4800
tgagcatggc agctggttgc tccatttgag agacacgctg gcgacacact ccgtccatcc 4860
gactgcccct gctgtgctgc tcaaggccac aggcacacag gtctcattgc ttctgactag 4920
attattattt gggggaactg gacacaatag gtctttctct cagtgaaggt ggggagaagc 4980
tgaaccggc 4989
<210> 4
<211> 25
<212> DNA
<213> synthetic construct
<400> 4
gcgcccgctg catgacgcct gcaac 25
<210> 5
<211> 24


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-6-
<212> DNA
<213> synthetic construct
<400> 5
cgggcggctc acctggagct ggcg 24
<210> 6
<211> 18
<212> DNA
<213> synthetic construct
<400> 6
aggcggctga cggcacta 18
<210> 7
<211> 19
<212> DNA
<213> synthetic construct
<400> 7
caggcgtcat gcagcgggc 19
<210> 8
<211> 25
<212> DNA
<213> synthetic construct
<400> 8
cggagatgcc gcatgccagc gcagg 25
<210> 9
<211> 18
<212> DNA
<213> synthetic construct
<400> 9
gacagcgtcg gagcgatc 18


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
<210> 10
<211> 18
<212> DNA
<213> synthetic construct
<400> 10
atctctccgc ttcctttc 18
<210> 11
<211> 18
<212> DNA
<213> synthetic construct
<400> 11
gaaaggaagc ggagagat 18
<210> 12
<211> 12
<212> DNA
<213> synthetic construct
<400> 12
ccggagccag ac 12
<210> 13
<211> 12
<212> DNA
<213> synthetic construct
<400> 13
cacaggcgca ag 12
<210> 14
<211> 8
<212> DNA
<213> synthetic construct


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
_g_
<400> 14
cccgcccc 8
<210> 15
<211> 15
<212> DNA
<213> synthetic construct
<400> 15
agcccccaca gcgag 15
<210> 16
<211> 12
<212> DNA
<213> synthetic construct
<400> 16
gccggagaga gc 12
<210> 17
<211> 13
<212> DNA
<213> synthetic construct
<400> 17
aacagaggca gca 13
<210> 18
<211> 13
<212> DNA
<213> synthetic construct
<400> 18
ggacagggac cag 13
<210> 19


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-9-
<211> 14
<212> DNA
<213> synthetic construct
<400> 19
cggcaagcac acag 14
<210> 20
<211> 15
<212> DNA
<213> synthetic construct
<400> 20
ggcaggcagg cacac 15
<210> 21
<211> 328
<212> PRT
<213> human
<400> 21
Met Leu Pro Arg Val Gly Cys Pro Ala Leu Pro Leu Pro Pro Pro Pro
1 5 10 15
Leu Leu Pro Leu Leu Pro Leu Leu Leu Leu Leu Leu Gly Ala Ser Gly
20 25 30
Gly Gly Gly Gly Ala Arg Ala Glu Val Leu Phe Arg Cys Pro Pro Cys
35 40 45
Thr Pro Glu Arg Leu Ala Ala Cys Gly Pro Pro Pro Val Ala Pro Pro
50 55 60
Ala Ala Val Ala Ala Val Ala Gly Gly Ala Arg Met Pro Cys Ala Glu
65 70 75 80
Leu Val Arg Glu Pro Gly Cys Gly Cys Cys Ser Val Cys Ala Arg Leu
85 90 95


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-10-
Glu Gly Glu Ala Cys Gly Val Tyr Thr Pro Arg Cys Gly Gln Gly Leu
100 105 110
Arg Cys Tyr Pro His Pro Gly Ser Glu Leu Pro Leu Gln Ala Leu Val
115 120 125
Met Gly Glu Gly Thr Cys Glu Lys Arg Arg Asp Ala Glu Tyr Gly Ala
130 135 140
Ser Pro Glu Gln Val Ala Asp Asn Gly Asp Asp His Ser Glu Gly Gly
145 150 155 160
Leu Val Glu Asn His Val Asp Ser Thr Met Asn Met Leu Gly Gly Gly
165 170 175
Gly Ser Ala Gly Arg Lys Pro Leu Lys Ser Gly Met Lys Glu Leu Ala
180 185 190
Val Phe Arg Glu Lys Val Thr Glu Gln His Arg Gln Met Gly Lys Gly
195 200 205
Gly Lys His His Leu Gly Leu Glu Glu Pro Lys Lys Leu Arg Pro Pro
210 215 220
Pro Ala Arg Thr Pro Cys Gln Gln Glu Leu Asp Gln Val Leu Glu Arg
225 230 235 240
Ile Ser Thr Met Arg Leu Pro Asp Glu Arg Gly Pro Leu Glu His Leu
245 250 255
Tyr Ser Leu His Ile Pro Asn Cys Asp Lys His Gly Leu Tyr Asn Leu
260 265 270
Lys Gln Cys Lys Met Ser Leu Asn Gly Gln Arg Gly Glu Cys Trp Cys
275 280 285
Val Asn Pro Asn Thr Gly Lys Leu Ile Gln Gly Ala Pro Thr Ile Arg
290 295 300
Gly Asp Pro Glu Cys His Leu Phe Tyr Asn Glu Gln Gln Glu Ala Cys
305 310 315 320


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-11-
Gly Val His Thr Gln Arg Met Gln
325
<210> 22
<211> 39
<212> PRT
<213> human
<400> 22
Met Leu Pro Arg Val Gly Cys Pro Ala Leu Pro Leu Pro Pro Pro Pro
1 5 10 15
Leu Leu Pro Leu Leu Pro Leu Leu Leu Leu Leu Leu Gly Ala Ser Gly
20 25 30
Gly Gly Gly Gly Ala Arg Ala
<210> 23
<211> 289
<212> PRT
<213> human
<400> 23
Glu Val Leu Phe Arg Cys Pro Pro Cys Thr Pro Glu Arg Leu Ala Ala
1 5 10 15
Cys Gly Pro Pro Pro Val Ala Pro Pro Ala Ala Val Ala Ala Val Ala
20 25 30
Gly Gly Ala Arg Met Pro Cys Ala Glu Leu Val Arg Glu Pro Gly Cys
35 40 45
Gly Cys Cys Ser Val Cys Ala Arg Leu Glu Gly Glu Ala Cys Gly Val
50 55 60
Tyr Thr Pro Arg Cys Gly Gln Gly Leu Arg Cys Tyr Pro His Pro Gly
65 70 75 80


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-12-
Ser Glu Leu Pro Leu Gln Ala Leu Val Met Gly Glu Gly Thr Cys Glu
85 90 95
Lys Arg Arg Asp Ala Glu Tyr Gly Ala Ser Pro Glu Gln Val Ala Asp
100 105 110
Asn Gly Asp Asp His Ser Glu Gly Gly Leu Val Glu Asn His Val Asp
115 120 125
Ser Thr Met Asn Met Leu Gly Gly Gly Gly Ser Ala Gly Arg Lys Pro
130 135 140
Leu Lys Ser Gly Met Lys Glu Leu Ala Val Phe Arg Glu Lys Val Thr
145 150 155 160
Glu Gln His Arg Gln Met Gly Lys Gly Gly Lys His His Leu Gly Leu
165 170 175
Glu Glu Pro Lys Lys Leu Arg Pro Pro Pro Ala Arg Thr Pro Cys Gln
180 185 190
Gln Glu Leu Asp Gln Val Leu Glu Arg Ile Ser Thr Met Arg Leu Pro
195 200 205
Asp Glu Arg Gly Pro Leu Glu His Leu Tyr Ser Leu His Ile Pro Asn
210 215 220
Cys Asp Lys His Gly Leu Tyr Asn Leu Lys Gln Cys Lys Met Ser Leu
225 230 235 240
Asn Gly Gln Arg Gly Glu Cys Trp Cys Val Asn Pro Asn Thr Gly Lys
245 250 255
Leu Ile Gln Gly Ala Pro Thr Ile Arg Gly Asp Pro Glu Cys His Leu
260 265 270
Phe Tyr Asn Glu Gln Gln Glu Ala Cys Gly Val His Thr Gln Arg Met
275 280 285
Gln


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
-13-
<210> 24
<211> 291
<212> PRT
<213> human
<400> 24
Met Gln Arg Ala Arg Pro Thr Leu Trp Ala Ala Ala Leu Thr Leu Leu
1 5 10 15
Val Leu Leu Arg Gly Pro Pro Val Ala Arg Ala Gly Ala Ser Ser Gly
20 25 30
Gly Leu Gly Pro Val Val Arg Cys Glu Pro Cys Asp Ala Arg Ala Leu
35 40 45
Ala Gln Cys Ala Pro Pro Pro Ala Val Cys Ala Glu Leu Val Arg Glu
50 55 60
Pro Gly Cys Gly Cys Cys Leu Thr Cys Ala Leu Ser Glu Gly Gln Pro
65 70 75 80
Cys Gly Ile Tyr Thr Glu Arg Cys Gly Ser Gly Leu Arg Cys Gln Pro
85 90 95
Ser Pro Asp Glu Ala Arg Pro Leu Gln Ala Leu Leu Asp Gly Arg Gly
100 105 110
Leu Cys Val Asn Ala Ser Ala Val Ser Arg Leu Arg Ala Tyr Leu Leu
115 120 125
Pro Ala Pro Pro Ala Pro Gly Asn Ala Ser Glu Ser Glu Glu Asp Arg
130 135 140
Ser Ala Gly Ser Val Glu Ser Pro Ser Val Ser Ser Thr His Arg Val
145 150 155 160
Ser Asp Pro Lys Phe His Pro Leu His Ser Lys Ile Ile Ile Ile Lys
165 170 175
Lys Gly His Ala Lys Asp Ser Gln Arg Tyr Lys Val Asp Tyr Glu Ser


CA 02376284 2001-12-05
WO 00/78341 PCT/AU00/00693
- 14-
180 185 190
Gln Ser Thr Asp Thr Gln Asn Phe Ser Ser Glu Ser Lys Arg Glu Thr
195 200 205
Glu Tyr Gly Pro Cys Arg Arg Glu Met Glu Asp Thr Leu Asn His Leu
210 215 220
Lys Phe Leu Asn Val Leu Ser Pro Arg Gly Val His Ile Pro Asn Cys
225 230 235 240
Asp Lys Lys Gly Phe Tyr Lys Lys Lys Gln Cys Arg Pro Ser Lys Gly
245 250 255
Arg Lys Arg Gly Phe Cys Trp Cys Val Asp Lys Tyr Gly Gln Pro Leu
260 265 270
Pro Gly Tyr Thr Thr Lys Gly Lys Glu Asp Val His Cys Tyr Ser Met
275 280 285
Gln Ser Lys
290