Note: Descriptions are shown in the official language in which they were submitted.
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PLANT FATTY ACID DESATURASES AND ALLELES THEREFOR
FIELD OF THE INVENTION
The invention is in the field of plant biology, involving compositions and
methods
related to fatty acid metabolism in plants. Aspects of the invention include
genes and
enzymes involved in fatty acid metabolism in plants, as well as plants and
plant parts
having the genes and expressing the enzymes, and methods for making the plants
and plant
parts using the genes (including recombinant genetic engineering methods and
classical
plant breeding methods using markers of the invention).
BACKGROUND OF THE INVENTION
Fatty acids are acyl lipids that are found in a variety of plant tissues,
including the
triacylglycerols in oil bodies of seeds and fruits, as well as the glycolipids
and
phospholipids in leaves, roots or shoots. Fatty acids include saturated and
unsaturated
monocarboxylic acids with unbranched even-numbered carbon chains, such as the
unsaturated fatty acids: oleic (18:1, i.e. a C18 chain with a double bond in
position 1),
linoleic (18:2) and linolenic (18:3) acid.
Significant efforts have been made to manipulate the fatty acid profile of
plants,
particularly oil-seed varieties such as canola that are used for the large-
scale production of
commercial fats and oils (see for example U.S. Patent Nos. 5,625,130 issued to
Grant et al.
29 April 1997; 5,668,299 issued to DeBonte et al. 16 September 1997; 5,767,338
issued to
Fan 16 June 1998; 5,777,201 issued to Poutre et al. 7 July 1998; 5,840,946
issued to Wong
et al. 24 November 1998; and 5,850,026 issued to DeBonte et al. 15 December
1998).
A reduction in the linolenic acid content of plant oils may be desirable for
some
applications. Low linolenic acid cultivars of B. napus have for example been
developed
from the cultivar Oro (Robbelen and Nitsch, 1975, L. ZPflanzenzCTchtg 75:93),
by
mutagenesis including the low linolenic acid cultivars Stellar (Scarth et al.,
1988, Can J
Plant Sci 68:509) and Apollo (Scarth et al., 1994, Can JPlant Sci 75:203). The
Apollo line
has been used to identify molecular markers associated with low linolenic acid
loci in a
double haploid population derived from a cross between the Apollo line (low
linolenic) and
a high linolenic line (YN90-1016), using random amplification of polymorphic
DNAs and
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bulk segregant analysis (Somers et al., 1998, Theoretical and Applied Genetics
96(6/7):897). The rapeseed fad3 gene, one of 13 markers identified by Somers
et al., supra,
was mapped near the locus controlling 14% of the variation in linolenic acid
content,
confirming a link between the fad3 gene and a low linolenic acid phenotype
(Jourdren et
al., 1996, Theoretical and Applied Genetics 93:512).
The product of the Fad3 gene is a fatty acid desaturase known variously as
delta-15
fatty acid desaturase, linoleic acid desaturase, omega-3 fatty acid
desaturase, Fad3 or 15-
DES (Arondel et al., 1992, Science 258:1353; Yadav et al., 1993, Plant
Physiol. 103:467;
WO 93/11245; and WO 98/56239 published 17 December 1998), hereinafter called
Fad3.
Fad 3 is involved in the enzymatic conversion of linoleic acid to alpha-
linolenic acid. In
WO 98/56239, DeBonte et al. disclose mutant Fad3 genes, and identify regions
of the Fad3
enzyme that are said to contain conserved amino acid motifs which may be
mutated to alter
fatty acid metabolism in a plant (see Tables 5 and 6 therein). The genomic
regions
identified by DeBonte et al. generally coincide with the first two of three
'Histidine Box'
motifs that have been imputed to have a role in the functional activity of the
Fad3 enzyme.
SUMMARY OF THE INVENTION
It has unexpectedly been discovered that plant fatty acid metabolism may be
altered
by mutations in the Fad3 enzyme, particularly by amino acid substitutions in
regions of the
protein outside of the regions taught to be functionally important in WO
98/56239. In one
aspect, the invention accordingly provides new variants of the Fad3 enzyme,
comprising
non-conserved amino acid substitutions, as well as nucleic acid sequences
encoding such
peptides. It is disclosed herein that plants having the Fad3 alleles of the
invention exhibit a
low linolenic acid phenotype. Accordingly, other aspects of the invention
include
transgenic plants and plant parts. As used herein, 'plant parts' includes
plant cells, seeds,
pollen bearing the nucleic acids of the invention or expressing the Fad3
enzymes of the
invention or having the Fad3 coding sequences of the invention. Vectors
capable of
transforming plant cells are provided, comprising the nucleic acids of the
invention,
including Fad3 coding sequences. Corresponding methods are provided for
obtaining the
transgenic plants of the invention. Methods are provided for using the plants
of the
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invention, including selected plants and transgenic plants, to obtain plant
products. As used
herein, "plant products" includes anything derived from a plant of the
invention, including
plant parts such as seeds, meals, fats or oils, including such plant products
having altered
linolenic acid concentrations. Amplification primers for identifying the Fad3
alleles of the
invention are provided, together with methods of obtaining plants using the
Fad3 alleles of
the invention as markers.
Marker assisted plant breeding programs are provided by the invention, wherein
the
Fad3 alleles of the invention, such as Fad3A and Fad3C, may be identified in
plant lines
subjected to selective breeding.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 is a listing of the amino acid sequence of the Fad3A protein from the
Apollo cultivar (SEQ ID NO: 1 ), showing positions of amino acid substitutions
in
accordance with various aspects of the invention, at positions 213, 275 and
347. One of the
prior-art-identified histidine box sequences, HDCGH, is also boxed for
reference.
Figure 2 is a pairwise alignment of the Apollo Fad3A ("ApolloA") and partial
Fad3C ("ApolloC") sequences with the derived Brassica napus omega-3 fatty acid
desaturase amino acid sequence which is GenBank accession number L22962 (SEQ
ID
N0:2), showing: Identities = 369/380 (97%), Positives = 372/380 (97%), Gaps =
3/380,
using the BLASTp program. In the Consensus sequence, two regions identified as
functionally important in WO 98/56239 appear in boxes. A putative 'histidine
box' within
the first of these regions, identified in the prior art relating to Fad3
enzymes, is also boxed
in the ApolloA and L22962 sequences.
Figure 3 a pairwise alignment of the Apollo Fad3A sequence and the derived
Brassica napus omega-3 fatty acid desaturase amino acid sequence which is
GenBank
accession number L01418 (SEQ ID N0:3), showing: Identities = 359/383 (93%),
Positives
= 368/383 (95%), Gaps = 3/383 (0%), using the BLASTp program.
Figure 4 is a pairwise alignment of the Apollo Fad3A sequence and the derived
Arabidopsis thaliana omega-3 fatty acid desaturase amino acid sequence which
is
GenBank accession numbers D 17579 and D26508 (SEQ ID N0:4), showing:
Identities =
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347/386 (89%), Positives = 361/386 (92%), Gaps = 6/386 (1%), using the BLASTp
program. Position 98 in the sequence is also highlighted, to provide a
reference point with
respect to the sequence shown in Figure 5 which begins at residue 98.
Figure 5 is a partial pairwise alignment of the Apollo Fad3A and Fad3C
sequences
and the derived YN90-1016 Fad3 sequence (SEQ ID NO:S).
Figure 6 is a partial pairwise alignment of the Apollo Fad3A sequence and the
derived N89-53 Fad3 sequence (SEQ ID N0:6).
Figure 7 shows an Apollo Fad3A cDNA sequence (SEQ ID N0:7) and a partial
Fad3C cDNA sequence, aligned.
Figure 8 is the Apollo Fad3A genomic DNA sequence (SEQ ID N0:8).
Figure 9 is a multiple protein sequence alignment, carried out using BLASTP
software, comparing the Apollo Fad3A sequence (SEQ ID NO:1) to a variety of
known
plant delta 15 fatty acid desaturase protein sequences (SEQ ID NO: 9 to SEQ ID
N0:42).
Figure 10 is a comparison of the partial genomic pFad3A (Apollo) and partial
genomic pFad3Y (YN90-1016) sequences, discussed in the Examples, with a
consensus
sequence shown between them. The pFad3A sequence is the top sequence, and
begins at
nucleotide 954 of the Apollo Fad3A genomic DNA sequence of Figure 8.
Figure 11 is a sequence alignment performed using the CLUSTALW program,
showing the alignment betweeen a genomic Fad3A sequence (SEQ ID N0:8) and a
partial
genomic Fad3C sequence.
DETAILED DESCRIPTION OF THE INVENTION
In one aspect, the invention provides recombinant nucleic acids encoding a
plant
fatty acid desaturase. By recombinant, it is meant herein that a nucleic acid
is not a
naturally occurring sequence, or it is a sequence that is made by an
artificial combination of
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two otherwise separated segments of nucleic acid sequence. Such combinations
of
sequences may be achieved by a wide variety of genetic engineering techniques,
such as
mutagenesis and site-specific-recombination of one or more nucleotides
(Beetham et al.,
1999, Proc. Natl. Acad. Sci. USA 96:8774; Zhu et al., 1999, Proc. Natl. Acad.
Sci. USA
96:87768). By fatty acid desaturase, it is meant herein that a protein
exhibits activity
manifested as the introduction of a double bond in the biosynthesis of a fatty
acid. For
example, Fad3 enzymes are defined by the activity of introducing the third
double bond in
the biosynthesis of 16:3 or 18:3 fatty acids.
In various aspects of the invention, the nucleic acid sequence of the
invention may
encode an amino acid substitution in the desaturase. By substitution, it is
meant that the
amino acid sequence is other than it would have been but for the recombination
of the
nucleic acid encoding the protein. The amino acid substitution may for example
be at a
position selected from the group consisting of amino acid positions
corresponding to amino
acid positions 213, 217, 224, 275, 281 and 347 of Apollo Fad3A (SEQ ID NO: 1).
By
'corresponding to', in comparison to the Apollo Fad3A (or Fad3C) sequence, it
is meant
that the positions are aligned when the sequences being compared are optimally
aligned,
for example using the BLASTP algorithm, with gaps permitted, and allowing for
conservative substitutions, as discussed further herein.
In alternative embodiments, amino acid substitutions in the desaturase may be
made
in particular motifs. For example, substitutions may be made within motifs,
such as the
motif STTCwszM centered on a position corresponding to position 213 of Apollo
Fad3A;
the motif STTCWSIMLATLVYLSFL COrreSpOridlrig t0 positions 210 to 227 of Apollo
Fad3A;
the motif sYLR~~L centered on a position corresponding to position 275 of
Apollo Fad3A;
the motif SXXXDHYVSD (in which X represents any amino acid) . beglrllllrig at
a
position corresponding to position 347 of Apollo Fad3A; a position in the
motif
STTCWSIMLAT corresponding to positions 210 to 220 of Apollo Fad3A; and, a
position in
the motif SYLRGGLTTIDRD CO1T'eSpOndlng to positions 272 to 284 of Apollo
Fad3A.
It is well known in the art that some modifications and changes can be made in
the
structure of a polypeptide without substantially altering the biological
function of that
peptide, to obtain a biologically equivalent polypeptide. As used herein, the
term
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"conserved amino acid substitutions" refers to the substitution of one amino
acid for
another at a given location in the peptide, where the substitution can be made
without any
appreciable loss or gain of function, to obtain a biologically equivalent
polypeptide. In
making such changes, substitutions of like amino acid residues can be made on
the basis of
relative similarity of side-chain substituents, for example, their size,
charge,
hydrophobicity, hydrophilicity, and the like, and such substitutions may be
assayed for
their effect on the function of the peptide by routine testing. Conversely, as
used herein, the
term "non-conserved amino acid substitutions" refers to the substitution of
one amino acid
for another at a given location in the peptide, where the substitution causes
an appreciable
loss or gain of function of the peptide, to obtain a polypeptide that is not
biologically
equivalent.
In some embodiments, conserved amino acid substitutions may be made where an
amino acid residue is substituted for another having a similar hydrophilicity
value (e.g.,
within a value of plus or minus 2.0), where the following hydrophilicity
values are assigned
to amino acid residues (as detailed in United States Patent No. 4,554,101,
incorporated
herein by reference): Arg (+3.0); Lys (+3.0); Asp (+3.0); Glu (+3.0); Ser
(+0.3); Asn
(+0.2); Gln (+0.2); Gly (0); Pro (-0.5); Thr (-0.4); Ala (-0.5); His (-0.5);
Cys (-1.0); Met (-
1.3); Val (-1.5); Leu (-1.8); Ile (-1.8); Tyr (-2.3); Phe (-2.5); and Trp (-
3.4). Non-conserved
amino acid substitutions may be made were the hydrophilicity value of the
residues is
significantly different, e.g. differing by more than 2Ø For example, on this
basis, the
following amino acid substitutions for the wild type Cys (-1.0) at a position
corresponding
to amino acid 213 in Apollo Fad3A would be non-conserved substitutions: Trp (-
3.4), Arg
(+3.0); Lys (+3.0); Asp (+3.0); Glu (+3.0). Similarly the following amino acid
substitutions for the wild type Arg (+3.0) at a position corresponding to
amino acid 275 in
Apollo Fad3A would be non-conserved substitutions: Ser (+0.3); Asn (+0.2); Gln
(+0.2);
Gly (0); Pro (-0.5); Thr (-0.4); Ala (-0.5); His (-0.5); Cys (-1.0); Met (-
1.3); Val (-1.5); Leu
(-1.8); Ile (-1.8); Tyr (-2.3); Phe (-2.5); and Trp (-3.4). Similarly the
following amino acid
substitutions for the wild type Ser (+0.3) at a position corresponding to
amino acid 347 in
Apollo Fad3A would be non-conserved substitutions: Arg (+3.0); Lys (+3.0); Asp
(+3.0);
Glu (+3.0); Leu (-1.8); Ile (-1.8); Tyr (-2.3); Phe (-2.5); and Trp (-3.4).
In alternative embodiments, conserved amino acid substitutions may be made
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where an amino acid residue is substituted for another having a similar
hydropathic index
(e.g., within a value of plus or minus 2.0). In such embodiments, each amino
acid residue
may be assigned a hydropathic index on the basis of its hydrophobicity and
charge
characteristics, as follows: Ile (+4.5); Val (+4.2); Leu (+3.8); Phe (+2.8);
Cys (+2.5); Met
(+1.9); Ala (+1.8); Gly (-0.4); Thr (-0.7); Ser (-0.8); Trp (-0.9); Tyr (-
1.3); Pro (-1.6); His
(-3.2); Glu (-3.5); Gln (-3.5); Asp (-3.5); Asn (-3.5); Lys (-3.9); and Arg (-
4.5). Non-
conserved amino acid substitutions may be made were the hydropathic index of
the
residues is significantly different, e.g. differing by more than 2Ø For
example, on this
basis, the following amino acid substitutions for the wild type Cys (+2.5) at
a position
corresponding to amino acid 213 in Apollo Fad3A would be non-conserved
substitutions:
Ile (+4.5); Gly (-0.4); Thr (-0.7); Ser (-0.8); Trp (-0.9); Tyr (-1.3); Pro (-
1.6); His (-3.2);
Glu (-3.5); Gln (-3.5); Asp (-3.5); Asn (-3.5); Lys (-3.9); and Arg (-4.5).
Similarly the
following amino acid substitutions for the wild type Arg (-4.5) at a position
corresponding
to amino acid 275 in Apollo Fad3A would be non-conserved substitutions: Ile
(+4.5); Val
(+4.2); Leu (+3.8); Phe (+2.8); Cys (+2.5); Met (+1.9); Ala (+1.8); Gly (-
0.4); Thr (-0.7);
Ser (-0.8); Trp (-0.9); Tyr (-1.3); Pro (-1.6). Similarly the following amino
acid
substitutions for the wild type Ser (-0.8) at a position corresponding to
amino acid 347 in
Apollo Fad3A would be non-conserved substitutions: Ile (+4.5); Val (+4.2); Leu
(+3.8);
Phe (+2.8); Cys (+2.5); Met (+1.9); Ala (+1.8); His (-3.2); Glu (-3.5); Gln (-
3.5); Asp (-
3.5); Asn (-3.5); Lys (-3.9); and Arg (-4.5). Similarly, the the following
amino acid
substitutions for the wild type Met (+1.9) at a position corresponding to
amino acid 217 in
Apollo Fad3A would be non-conserved substitutions: Ile (+4.5); Val (+4.2); Gly
(-0.4); Thr
(-0.7); Ser (-0.8); Trp (-0.9); Tyr (-1.3); Pro (-1.6); His (-3.2); Glu (-
3.5); Gln (-3.5); Asp (-
3.5); Asn (-3.5); Lys (-3.9); and Arg (-4.5). Similarly, the the following
amino acid
substitutions for the wild type Leu (+3.8) at a position corresponding to
amino acid 224 in
Apollo Fad3A would be non-conserved substitutions: Ala (+1.8); Gly (-0.4); Thr
(-0.7); Ser
(-0.8); Trp (-0.9); Tyr (-1.3); Pro (-1.6); His (-3.2); Glu (-3.5); Gln (-
3.5); Asp (-3.5); Asn
(-3.5); Lys (-3.9); and Arg (-4.5).
In alternative embodiments, conserved amino acid substitutions may be made
where an amino acid residue is substituted for another in the same class,
where the amino
acids are divided into non-polar, acidic, basic and neutral classes, as
follows: non-polar:
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Ala, Val, Leu, Ile, Phe, Trp, Pro, Met; acidic: Asp, Glu; basic: Lys, Arg,
His; neutral: Gly,
Ser, Thr, Cys, Asn, Gln, Tyr. Non-conserved amino acid substitutions may be
made were
the residues do not fall into the same class, for example substitution of a
basic amino acid
for a neutral or non-polar amino acid.
In alternative aspects of the invention, mutant plant fatty acid desaturases,
such as
Fad3 enzymes, are provided that have amino acid substitutions corresponding to
the
substitutions found in the Apollo Fad3A or Fad3C proteins: Ala substituted in
position 213,
or Cys substituted in position 275, or Arg substituted in position 347, or Val
substituted in
position 217, or Pro substituted in position 224, or Val substituted in
position 281. In
alternative embodiments, amino acid substitutions may be made at these
positions that are
at least as non-conserved as the substitutions found in Apollo Fad3A or Fad3C.
For
example, the substitution of Ala for Cys at position 213 of Apollo Fad3A
constitutes a
change on the foregoing hydrophilicity scale of -1.0 to -0.5, i.e. a
difference of 0.5.
Substitutions of similar magnitude of change would comprise substituting any
one of the
following amino acids for Cys (-1.0): Arg (+3.0); Lys (+3.0); Asp (+3.0); Glu
(+3.0); Ser
(+0.3); Asn (+0.2); Gln (+0.2); Gly (0); Pro (-0.5); Thr (-0.4); Ala (-0.5);
His (-0.5); Val (-
1.5); Leu (-1.8); Ile (-1.8); Tyr (-2.3); Phe (-2.5); and Trp (-3.4).
Similarly, the substitution
of Arg for Ser at position 347 of Apollo Fad3A constitutes a change on the
foregoing
hydrophilicity scale of +3.0 to +0.3, i.e. a difference of 2.7. Substitutions
of similar
magnitude of change would comprise substituting any one of the following amino
acids for
Ser (+0.3): Phe (-2.5); and Trp (-3.4).
In alternative embodiments, using amino acid substitutions based on the
foregoing
hydropathic index scale, the substitution of Ala for Cys at position 213 of
Apollo Fad3A
constitutes a change on the foregoing hydrophilicity scale of +2.5 to +1.8,
i.e. a difference
of 0.7. Substitutions of similar magnitude of change would comprise
substituting any one
of the following amino acids for Cys (+2.5): Gly (-0.4); Thr (-0.7); Ser (-
0.8); Trp (-0.9);
Tyr (-1.3); Pro (-1.6); His (-3.2); Glu (-3.5); Gln (-3.5); Asp (-3.5); Asn (-
3.5); Lys (-3.9);
and Arg (-4.5); Ile (+4.5); Val (+4.2); Leu (+3.8). Similarly, the
substitution of Cys for Arg
at position 275 of Apollo Fad3A constitutes a change on the foregoing
hydropathic index
of -4.5 to +2.5, i.e. a difference of 7Ø Substitutions of similar magnitude
of change would
comprise substituting any one of the following amino acids for Arg (-4.5): Ile
(+4.5); Val
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(+4.2); Leu (+3.8); Phe (+2.8). Similarly, the substitution of Arg for Ser at
position 347 of
Apollo Fad3A constitutes a change on the foregoing hydropathic index of -0.8
to -4.5, i.e. a
difference of 3.7. Substitutions of similar magnitude of change would comprise
substituting
any one of the following amino acids for Ser (-0.8): Ile (+4.5); Val (+4.2);
Leu (+3.8).
One aspect of the invention is the recognition of functionally important
sequence
motifs in plant delta 1 S fatty acid desaturases, particularly the motifs in
the conserved
regions that surround the amino acid substitutions in the Apollo Fad3
proteins: including
the motif sTTCwsiM centered on position 213; the motif sYLR~~L centered on
position 275;
and the motif SXXXDHYVSD beginning at position 347. Non-conservative amino
acid
substitutions within these motifs of plant delta 15 fatty acid desaturases are
an aspect of the
present invention. Plant delta 15 fatty acid desaturases having such non-
conserved
substitutions may be useful in transgenic plants of the invention to alter
fatty acid
metabolism, particularly the fatty acid composition of seed oils.
In various aspects, the invention provides isolated nucleic acid and protein
sequences. By isolated, it is meant that the isolated substance has been
substantially
separated or purified away from other biological components with which it
would other
wise be associated, for example in vivo. The term 'isolated' therefore
includes substances
purified by standard purification methods, as well as substances prepared by
recombinant
expression in a host, as well as chemically synthesized substances.
The invention provides vectors comprising nucleic acids of the invention. A
vector
is a nucleic acid molecule that may be introduced into a host cell, to produce
a transformed
host cell. A vector may include nucleic acid sequences that permit it to
replicate in the host
cell, such as an origin of replication. A vector may also include one or more
selectable
marker genes and other genetic elements known in the art. A transformed cell
is a cell into
which has been introduced a nucleic acid molecule by molecular biology
techniques. As
used herein, the term transformation encompasses all such techniques by which
a nucleic
acid molecule might be introduced into a host cell, including transformation
with
Agrobacterium vectors, transfection with viral vectors, transformation with
plasmid vectors
and introduction of naked DNA by electroporation, lipofection and particle gun
acceleration.
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In one aspect the invention provides amplification primers or probes that may
be
used to identify Fad3 nucleic acid sequences of the invention, such as the
Apollo Fad3A or
Fad3C nucleic acid sequences, from other nucleic acid sequences. As used
herein, the term
"Apollo Fad3 nucleic acid sequences", means the naturally occurring nucleic
acid
sequences, and portions thereof, encoding the Apollo Fad3 enzyme, including
Fad3A and
Fad3C. For example, primers or probes may be synthesized that are
complimentary to
portions of the Apollo microsomal Fad3A or Fad3C alleles that differ from the
sequence of
the Fad3 allele reported by Yadav et al. 1993, Plant Physiology 103:467. An
example of
such a primer is described in Example 1, wherein one of the selected primers
is shown to
be capable of distinguishing plants having high linolenic acid content from
plants having
low linolenic acid content. Such primers or probes may comprise 5 or more
contiguous
residues complimentary to a Fad3 nucleic acid sequence of the invention, such
as Fad3A or
Fad3C. In some embodiments, the isolated nucleic acid probe or primer may be
capable of
hybridizing to a characteristic portion of the recombinant nucleic acid (i.e.
a part of the
recombinant sequence which differs from other sequences, such as wild type
sequences),
under selective hybridization conditions. Selective hybridization of this sort
may be used to
identify a Fad3 nucleic acid sequence of the invention.
In one aspect, the invention provides amplification primers that may be used
to
incorporate a sequence polymorphism into an amplified nucleic acid sequence,
such that a
novel restriction site is produced. For example, primers may be synthesized
that are
substantially complementary to portions of an allele of interest, but differ
from the
sequence by one or more point mutations that introduce a restriction enzyme
cleavage site
(Michaels et al., 1998, The Plant Journal 14(3): 381-385, and Neff et al.,
1998, The Plant
Journal 14(3): 387-392; both of which are incorporated herein by reference).
Primers such
as those described in Example 1, may be adapted to produce by amplification a
nucleic acid
that contains a restriction enzyme site that is unique to an allele. The
restriction site may be
cleaved by a restriction endonuclease to provide sequence information from
allele-specific
polymorphisms.
One aspect of the invention comprises a method of selecting plants, such as
Brassica napus seedlings, having a low linolenic acid content by utilizing PCR
primers to
CA 02386111 2002-03-28
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selectively amplify a desired Fad3 allele. This method may be used, for
example, to ensure
that selected progeny carry a desired allele conferring a low linolenic acid
oil phenotype.
In accordance with an embodiment of the method, seedlings of a first
segregating
backcross population, may be subjected to PCR analysis to detect the mutant
Fad3 nucleic
acid, and the selected plants backcrossed again to a recurrent parental line.
The
backcrossing and PCR analysis of the first seedling population may, for
example, proceed
through at least two more cycles to create a third segregating backcross
seedling
population, which may be self pollinated to create a third seedling
population. The third
seedling population may be subjected to PCR analysis for the mutant Fad3
nucleic acid,
and homozygotes may be selected for further pedigree breeding, such as
breeding of an
elite, low linolenic acid content strain.
In various embodiments, the invention comprises plants expressing the
desaturases
of the invention. In some embodiments, such plants will exhibit altered fatty
acid content in
one or more tissues. These aspects of the invention relate to all higher
plants, including
monocots and dicots, such as species from the genera Fragaria. Lotus,
Medicago,
Onobrychis, Triforium, Trigonelia, Vigna, Citrus, Linum. Geranium, Manihot,
Caucus,
Arabidopsis, Brassica, Raphanus, Sinapis, Atropa, Capsicum, Hyoscyamus,
Lycopersicon,
Nicotiana, Solanum, Petunia, Digitalis, Majorana, Cichorium, Helianthus,
Lactuca,
Bromus, Asparagus, Antirrhinum, Heterocatlis, Nemesia, Pelargonium, Panicum,
Penniserum, Ranunculus, Senecio, Salpiglossis, Cucarnis, Browallia, Glycine,
Lolium,
Zea, Triticum, Sorghum, and Datura. Such plants may include maize, wheat,
rice, barley,
soybean, beans, rapeseed, canola, alfalfa, flax, sunflower, cotton, clover,
lettuce, tomato
cucurbits, potato carrot, radish, pea lentils, cabbage, broccoli, brussel
sprouts, peppers,
apple, pear, peach, apricot, carnations and roses. More specifically, in
alternative
embodiments, plants for which the invention may be used in modifying fatty
acid content
include oil crops of the Cruciferae family: canola, rapeseed (Brassica spp.),
crambe
(Crambe spp.), honesty (Lunaria spp.) lesquerella (Lesquerela spp.), and
others; the
Composirae family: sunflower (Helianthus spp.), safflower (Carthamus spp.),
niger
(Guizotia spp.) and others; the Palmae family: palm (Elaeis spp.), coconut
(Cocos spp.)
and others; the Leguminosae family: peanut (Arachis spp.), soybean (Glycine
spp.) and
others; and plants of other families such as maize (Zea spp.), cotton
(Gossypium sp.),
jojoba (Simonasia sp.), flax (Linum sp.), sesame (Sesamum spp.), castor bean
(Ricinus
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spp.), olive (Olea spp.), poppy (Papaver spp.), spurge (Euphorbia, spp.),
meadowfoam
(Limnanthes spp.), mustard (Sinapis spp.) and cuphea (Cuphea spp.).
In some aspects of the invention, nucleic acids encoding novel Fad3 proteins
may
be introduced into plants by transformation, and expression of such nucleic
acids may be
mediated by promoters to which such coding sequences are operably linked. One
aspect of
the invention comprises plants transformed with nucleic acid sequences
encoding the fatty
acid desaturases of the invention. Transformation may for example be carried
out as
described in WO 94/11516, which is hereby incorporated by reference. In the
context of the
present invention, "promoter" means a sequence sufficient to direct
transcription of a gene
when the promoter is operably linked to the gene. The promoter is accordingly
the portion
of a gene containing DNA sequences that provide for the binding of RNA
polymerase and
initiation of transcription. Promoter sequences are commonly, but not
universally, located
in the 5' non-coding regions of a gene. A promoter and a gene are "operably
linked" when
such sequences are functionally connected so as to permit gene expression
mediated by the
promoter. The term "operably linked" accordingly indicates that DNA segments
are
arranged so that they function in concert for their intended purposes, such as
initiating
transcription in the promoter to proceed through the coding segment of a gene
to a
terminator portion of the gene. Gene expression may occur in some instances
when
appropriate molecules (such as transcriptional activator proteins) are bound
to the
promoter. Expression is the process of conversion of the information of a
coding sequence
of a gene into mRNA by transcription and subsequently into polypeptide
(protein) by
translation, as a result of which the protein is said to be expressed. As the
term is used
herein, a gene or nucleic acid is "expressible" if it is capable of expression
under
appropriate conditions in a particular host cell.
For the present invention, promoters may be used that provide for preferential
gene
expression within a specific organ or tissue, or during a specific period of
development. For
example, promoters may be used that are specific for embryogenesis (U.S.
Patent No.
5,723,765 issued 3 March 1998 to Oliver et al. ). Such promoters may, in some
instances,
be obtained from genomic clones of cDNAs. Depending upon the application of
the present
invention, those skilled in this art may choose a promoter for use in the
invention which
provides a desired expression pattern. Promoters may be identified from genes
which have
a differential pattern of expression in a specific tissue by screening a
tissue of interest, for
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WO 01/25453 PCT/CA00/01140
example, using methods described in United States Patent No. 4,943,674 and
European
Patent Application EP-A 0255378.
Various aspects of the present invention encompass nucleic acid or amino acid
sequences that are homologous to other sequences. As the term is used herein,
an amino
acid or nucleic acid sequence is "homologous" to another sequence if the two
sequences are
substantially identical and the functional activity of the sequences is
conserved (for
example, both sequences function as or encode a Fad3; as used herein, sequence
conservation or identity does not infer evolutionary relatedness). Nucleic
acid sequences
may also be homologous if they encode substantially identical amino acid
sequences, even
if the nucleic acid sequences are not themselves substantially identical, for
example as a
result of the degeneracy of the genetic code.
Two amino acid or nucleic acid sequences are considered substantially
identical if,
when optimally aligned, they share at least about 70% sequence identity. In
alternative
embodiments, sequence identity may for example be at least 75%, at least 90%
or at least
95%. Optimal alignment of sequences for comparisons of identity may be
conducted using
a variety of algorithms, such as the local homology algorithm of Smith and
Waterman,1981, Adv. Appl. Math 2: 482, the homology alignment algorithm of
Needleman
and Wunsch, 1970, J. Mol. Biol. 48:443, the search for similarity method of
Pearson and
Lipman, 1988, Proc. Natl. Acad Sci. USA 85: 2444, and the computerized
implementations
of these algorithms (such as GAP, BESTFIT, FASTA and TFASTA in the Wisconsin
Genetics Software Package, Genetics Computer Group, Madison, WI, U.S.A.).
Sequence
identity may also be determined using the BLAST algorithm, described in
Altschul et al.,
1990, J. Mol. Biol. 215:403-10 (using the published default settings).
Software for
performing BLAST analysis may be available through the National Center for
Biotechnology Information (through the Internet at
http://www.ncbi.nlm.nih.~ovn. The
BLAST algorithm involves first identifying high scoring sequence pairs (HSPs)
by
identifying short words of length W in the query sequence that either match or
satisfy some
positive-valued threshold score T when aligned with a word of the same length
in a
database sequence. T is referred to as the neighbourhood word score threshold.
Initial
neighbourhood word hits act as seeds for initiating searches to find longer
HSPs. The word
hits are extended in both directions along each sequence for as far as the
cumulative
alignment score can be increased. Extension of the word hits in each direction
is halted
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WO 01/25453 PCT/CA00/01140
when the following parameters are met: the cumulative alignment score falls
off by the
quantity X from its maximum achieved value; the cumulative score goes to zero
or below,
due to the accumulation of one or more negative-scoring residue alignments; or
the end of
either sequence is reached. The BLAST algorithm parameters W, T and X
determine the
S sensitivity and speed of the alignment. The BLAST program may use as
defaults a word
length (W) of 11, the BLOSUM62 scoring matrix (Henikoff and Henikoff, 1992,
Proc.
Natl. Acad Sci. USA 89: 1091 S-10919) alignments (B) of 50, expectation (E) of
10 (or 1 or
0.1 or 0.01 or 0.001 or 0.0001 ), M=S, N=4, and a comparison of both strands.
One measure
of the statistical similarity between two sequences using the BLAST algorithm
is the
smallest sum probability (P(N)), which provides an indication of the
probability by which a
match between two nucleotide or amino acid sequences would occur by chance. In
alternative embodiments of the invention, nucleotide or amino acid sequences
are
considered substantially identical if the smallest sum probability in a
comparison of the
test sequences is less than about 1, preferably less than about 0.1, more
preferably less than
about 0.01, and most preferably less than about 0.001.
An alternative indication that two nucleic acid sequences are substantially
identical
is that the two sequences hybridize to each other under moderately stringent,
or preferably
stringent, conditions. Hybridisation to filter-bound sequences under
moderately stringent
conditions may, for example, be performed in 0.5 M NaHP04, 7% sodium dodecyl
sulfate
(SDS), 1 mM EDTA at 65°C, and washing in 0.2 x SSC/0.1% SDS at
42°C (see Ausubel,
et al. (eds), 1989, Current Protocols in Molecular Biology, Vol. 1, Green
Publishing
Associates, Inc., and John Wiley & Sons, Inc., New York, at p. 2.10.3).
Alternatively,
hybridization to filter-bound sequences under stringent conditions may, for
example, be
performed in 0.5 M NaHP04, 7% SDS, 1 mM EDTA at 65 °C, and washing in
0.1 x
SSC/0.1% SDS at 68°C (see Ausubel, et al. (eds), 1989, supra).
Hybridization conditions
may be modified in accordance with known methods depending on the sequence of
interest
(see Tijssen, 1993, Laboratory Techniques in Biochemistry and Molecular
Biology --
Hybridization with Nucleic Acid Probes, Part I, Chapter 2 "Overview of
principles of
hybridization and the strategy of nucleic acid probe assays", Elsevier, New
York).
Generally, stringent conditions are selected to be about 5 °C lower
than the thermal melting
point for the specific sequence at a defined ionic strength and pH.
An alternative indication that two amino acid sequences are substantially
identical
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WO 01/25453 PCT/CA00/01140
is that one peptide is specifically immunologically reactive with antibodies
that are also
specifically immunoreactive against the other peptide. Antibodies are
specifically
immunoreactive to a peptide if the antibodies bind preferentially to the
peptide and do not
bind in a significant amount to other proteins present in the sample, so that
the preferential
binding of the antibody to the peptide is detectable in an immunoassay and
distinguishable
from non-specific binding to other peptides. Specific immunoreactivity of
antibodies to
peptides may be assessed using a variety of immunoassay formats, such as solid-
phase
ELISA immunoassays for selecting monoclonal antibodies specifically
immunoreactive
with a protein (see Harlow and Lane (1988) Antibodies, A Laboratory Manual,
Cold Spring
Harbor Publications, New York).
As used herein to describe nucleic acid or amino acid sequences the term
"heterologous" refers to molecules or portions of molecules, such as DNA
sequences, that
are artificially introduced into a particular host cell. Heterologous DNA
sequences may for
example be introduced into a host cell by transformation. Such heterologous
molecules
may include sequences derived from the host cell. Heterologous DNA sequences
may
become integrated into the host cell genome, either as a result of the
original
transformation of the host cells, or as the result of subsequent recombination
events.
In accordance with various aspects of the invention, plant cells may be
transformed
with heterologous nucleic acids. In this context, "heterologous" denotes any
nucleic acid
that is introduced by transformation. Transformation techniques that may be
employed
include plant cell membrane disruption by electroporation, microinjection and
polyethylene
glycol based transformation (such as are disclosed in Paszkowski et al. EMBO
J. 3:2717
(1984); Fromm et al., Proc. Natl. Acad. Sci. USA 82:5824 (1985); Rogers et
al., Methods
Enzymol. 118:627 (1986); and in U.S. Patent Nos. 4,684,611; 4,801,540;
4,743,548 and
5,231,019), biolistic transformation such as DNA particle bombardment (for
example as
disclosed in Klein, et al., Nature 327: 70 (1987); Gordon-Kamm, et al. "The
Plant Cell"
2:603 (1990); and in U.S. Patent Nos. 4,945,050; 5,015,580; 5,149,655 and
5,466,587);
Agrobacterium-mediated transformation methods (such as those disclosed in
Horsch et al.
Science 233: 496 (1984); Fraley et al., Proc. Nat'l Acad. Sci. USA 80:4803
(1983); and
U.S. Patent Nos. 4,940,838 and 5,464,763).
Transformed plant cells may be cultured to regenerate whole plants having the
CA 02386111 2002-03-28
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transformed genotype and displaying a desired phenotype, as for example
modified by the
expression of a heterologous Fad3 during growth or development. A variety of
plant
culture techniques may be used to regenerate whole plants, such as are
described in
Gamborg and Phillips, "Plant Cell, Tissue and Organ Culture, Fundamental
Methods",
Springer Berlin, 1995); Evans et al. "Protoplasts Isolation and Culture",
Handbook of
Plant Cell Culture, Macmillian Publishing Company, New York, 1983; or Binding,
"Regeneration of Plants, Plant Protoplasts", CRC Press, Boca Raton, 1985; or
in Klee et
al., Ann. Rev. ofPlantPhys. 38:467 (1987).
Standard techniques may be used for plant transformation, such as
transformation
of Arabidopsis. For example, wild type (WT) A. thaliana seeds of ecotype
"Columbia"
may be planted in 4" pots containing soil and plants grown in a controlled
growth chamber
or greenhouse. The vacuum infiltration method of in planta transformation
(Bechtold et al.,
1993) may be used to transform A. thaliana plants with overnight culture of A.
tumefaciens
strain GV3101 bearing both the helper nopoline plasmid and the binary
construct
containing the described chimeric gene. pMP90 is a disarmed Ti plasmid with
intact vir
region acting in traps, gentamycin and kanamycin selection markers as
described in Koncz
and Schell (1986). Following infiltration, plants may be grown to maturity and
seeds (T1)
collected from each pod individually. Seeds may be surface-sterilized and
screened on
selective medium containing 50 mg/L kanamycin with or without 200-300 mg/L
timentin.
After about four weeks on selection medium, the non-transformed seedlings will
generally
die. The transformed seedlings may be transferred to soil in pots. Leaf DNA
may be
isolated (Edwards et al., 1991 ) and analyzed by PCR for the presence of the
DNA insertion.
Genomic DNA may also be isolated and used in Southern hybridization (Southern,
1975)
to determine the copy number of the inserted sequence in a given transformant.
To
determine the segregation, T2 seeds may be collected from T1 plants.
Alternative embodiments of the invention may make use of techniques for
transformation of Brassica. Such as transformation of B. napus cv. Westar and
B.
carinata cv. Dodolla by co-cultivation of cotyledonary petioles or hypocotyl
explants with
A. tumefaciens bearing the plasmids described herein. Transformation of B.
napus plants
may, for example, be performed according to the method of Moloney et al.,
1989, Plant
Cell Rep 8: 238. Modifications of that method may include the introduction of
a 7-day
explant-recovery period following co-cultivation, on MS medium with the
hormone
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benzyladenine (BA), and the antibiotic timentin for the elimination of
Agrobacterium.
Transformation of B. carinata plants may be performed according to the method
by Babic
et al., 1998, Plant Cell Rep 17: 183. Cotyledonary petiole explants may be
dipped in
suspension of Agrobacterium bearing the desired constructs and placed on 7-cm
filter paper
(Whatman no. 1 ) on top of the regeneration medium for 2 days. After co-
cultivation,
explants may be transferred onto the selection medium containing 50 mg/L
kanamycin.
Regenerated green shoots may first be transferred to a medium to allow
elongation and
then to a rooting medium all containing 50 mg/L kanamycin. Putative
transformants with
roots (TO) may be transferred to soil. Genomic DNA may be isolated from
developing
leaves for PCR and Southern analyses. Seeds (T1) from transgenic plants may
then be
harvested. Transgenic plants may be observed and characterized for alteration
of traits,
particularly fatty acid content, and more particularly fatty acid content of
seed oils.
Example 1: Isolation of Apollo Fad3 Sequences
Cloning and sequence analysis of the Fad3A gene is described below, the Fad3C
gene from Apollo was cloned and characterized in a similar manner.
PCR primers described in a publication by Jourdren et al. (1996) were used to
amplify the microsomal delta-15 fatty acid desaturase coding sequences (Fad3)
from the
following B. napus accessions: low linolenic acid variety Apollo (Scarth et
al. 1994) and
normal linolenic acid breeding lines YN90-1016 and N89-53 (Agriculture and
Agri-Food
Canada). The PCR reaction conditions used are described in Somers et al.,
1998, Theor.
Appl. Genet. 96: 897. The primer sequences were degenerate and named FAD3L and
FAD3R (see Table 1). An amplified DNA fragment was cloned from each accession
into
pGEM (Promega Corp, Madison WI, USA) and each of the clones (pFad3A, from
Apollo;
pFAD3Y, from YN90-1016; and pFad3N89 from N89-53) was sequenced using the di-
deoxy terminator cycle sequencing technique. The initial clones containing the
Fad3
coding sequence were lacking the 3' and 5' coding sequences. The 3' end of the
genomic
sequence from Apollo was PCR amplified using a primer (A047F, Table 1)
designed from
the pFad3A clone and a primer (A047R, Table 1) derived from the terminus of
the
genebank sequence L01418, a B. napus microsomal Fad3 gene. The 5' end of the
genomic
sequence from Apollo was PCR amplified using a primer (A046F, Table 1)
designed from
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the pFad3A clone and a primer (A046R, Table 1 ) derived from the terminus of
the
genebank sequence L01418. The Fad3 genomic DNA sequences were then aligned
with
genebank sequence L01418 (cDNA) and based on this alignment, the Apollo, YN90-
1016
and N89-53 Fad3 coding and non-coding sequences were distinguished, and the
coding
frame determined.
The three B. napus Fad3 coding sequences were converted to amino acid
sequences
using Lasergene, DNA STAR software and the protein sequences were aligned with
the
protein sequence derived from L01418. Differences at the protein sequence
level between
pFad3A and L01418, pFad3Y, pFad3N89 correlated to differences in the DNA
coding
sequence.
An alignment of the genomic DNA sequences in pFad3A, pFad3Y and pFad3N89
revealed several sequence differences within intron regions. PCR primers were
derived
from the pFad3A intron sequences and included the observed sequence
polymorphisms
(Table 1 ). DNA was extracted from many other oilseed accessions and these are
described
in Table 2.
Table 1. PCR primer sequences derived from the sequence of pFad3A
Primer name Sequence (5'-3') SEQ ID NO's
A006R AAG AGT GGC CAA CAT GAT CG 43
A007F ATT CTT AGC ATC TGC CTC G 44
A027F CCC CTT CTG AAT ACT GCG GT 45
A028F TTC CGG TAA TCC CCC TCT CA 46
A029R ACT GTA GTC ATC CCC AAA CAA AT 47
A036F GCA TCA AAA TCT TTA GCA TCG AA 48
A037F GGT GCA TGT TAG CAA ACA GTA AT 49
A046F CAT TTC ACT CAG AGC CCA CAC 50
A046R GAC CAA CGC CAG TAT TCA GA 51
A047F ATT ACG GGA TCT TCA ACA ACC A 52
A047R TAA AAA CAA CCAGAA ATA AGTAAA 53
A048 CTA TCA ATA GTTGTT AAT CCTCCA CA
54
A050 TTG GAC GAC CACTTG TCA GATT 55
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FAD3L GTG GAC ATG GGA GTT TYT CNG A 56
FAD3R TGG CAT CGA CCA ART GRT ART G 57
The pFad3A genomic DNA sequence is 3007 by (Fig. 8) and includes the entire
coding sequence. The pFad3A and pFad3Y (1864 bp) sequences were aligned and
there
were several sequence polymorphisms observed throughout the sequences (Figure
10). A
number of polymorphisms are further exemplified herein, centered at
nucleotides 191, 270,
693 and 1267 of pFad3A as shown in Fig. 10.
PCR primers that included sequence polymorphisms observed in the Apollo Fad3
coding sequences were designed from the pFad3A sequence (primers A028F, A029R,
A036F, A037F shown in Table 1). These primers were paired with different
conserved
PCR primers (designated A006R, A007F and A027F in Table 1) to demonstrate the
ability
to selectively amplify the Apollo Fad3 allele over other alleles, particularly
wild-type
alleles such as the YN90-1016 Fad3 allele. A DNA fragment of the predicted
size was
amplified from the Apollo DNA template in each case and was not amplified from
the
YN90-1016 DNA template. Therefore, the sequence polymorphisms observed in the
Apollo Fad3 gene may be used to selectively amplify and detect the mutant Fad3
allele
from Apollo. Similar sequence alignments of the Apollo Fad3 allele to other
crucifer
oilseed Fad3 alleles may be routinely used to identify sequence polymorphisms
that may
be used as a basis for the selective amplification of the Apollo Fad3 allele.
The alignment of pFad3A, pFad3Y and pFad3N89 with the Fad3 Genebank
sequence L01418 showed the position of introns and exons within pFad3A, pFad3Y
and
pFad3N89. The intron sequences were edited out to identify the coding sequence
of
pFad3A (852 by in length) to be aligned with the coding sequence of pFad3Y
(657 by in
length), showing a number of nucleotide polymorphisms (Fig. 10).
Both the pFad3A and pFad3Y coding sequences were converted to amino acid
sequences and aligned (Fig. 5). A non-conserved change (mutation) in the amino
acid
sequence between these protein sequences was identified at amino acid 275 of
the Apollo
Fad3 sequence (Apollo, cysteine; YN90-1016, arginine). Figure 9 shows the
extent to
which this mutation distinguishes the Apollo Fad3 enzyme from a very wide
variety of
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other known delta-15 fatty acid desaturases. Similarly, Figure 9 shows a
number of other
amino acid substitutions in the Apollo Fad3 sequence compared to other delta-
15 fatty acid
desaturases.
Identifying DNA sequence differences and primers.
The mutation at amino acid 275 (cysteine) is due to a single base pair
mutation,
shown boxed in Figure 7 (cDNA) at nucleotide 823, boxed in Figure 8 at
nucleotide 2685
and at corresponding nucleotide 1734 of the pFad3A DNA sequence of Figure 10
(the
pFad3A sequence of Figure 10 begins at nucleotide 954 of Figure 8). The wild
type
L01418, YN90-1016 and N89-53 Fad3 alleles all included a CGT (arginine) codon
and the
mutant Apollo Fad3 allele includes a TGT (cysteine) codon (Fig. 9).
A PCR primer (A048, Table 1) was designed to include the DNA sequence
polymorphism at a nucleotide corresponding to nucleotide 1734 of pFad3A (Fig.
10) where
the final nucleotide in the 3' end of the primer included an 'A' (Adenine)
nucleotide to
selectively PCR amplify the mutant Apollo Fad3 allele over corresponding
wildtype Fad3
alleles.
Specificity of selective amplification of Apollo microsomal Fad3A allele.
The mutant microsomal Fad3 alleles of Apollo are thought to be derived from a
low
linolenic acid mutant line from Germany, designated 'M11' (Robbelen G, Nitsch
A, 1975,
L. ZPflanzenzl7chtg 75:93). Amplification products indicative of the Apollo
Fad3A allele
were obtained using primers A048 and A050 (Table 1). A collection of genotypes
were
tested, as listed in table 2, for the presence of the C to T nucleotide
polymorphism of the
Apollo Fad3A allele. PCR amplification from an Apollo DNA template was also
assayed
as a control. Apart from Apollo, the only other genotypes showing the presence
of the
amplification product from the Apollo Fad3A gene included T097-3414, S86-69
and
Stellar. Stellar is the first spring canola quality B. napus variety developed
carrying low
linolenic acid and was derived from crosses with M11 (low linolenic acid)
(Scarth et al.
1988). Accession S86-69 is a low linolenic acid B. napus line selected from
the variety
Apollo. T097-3414 is a (BC3F4) B. juncea accession derived from interspecific
crosses of
B. juncea with S86-69 and selection for low linolenic acid. Therefore, all of
the accessions
showing amplification of the mutant Apollo Fad3A allele are related to Apollo,
in the sense
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that they are all descended from B. napus line M11 (by "descended from" it is
meant that a
plant is derived from another by methods of classical plant breeding,
including crossing
parent plant lines or self crossing of parent plants, but this does not
include methods of
genetic engineering in which nucleic acid sequences are recombined to produce
new
strains). Such PCR tests may be highly specific, and may be used in one aspect
of the
invention as a selective amplification assay for the presence of the Apollo
Fad3A or Fad3C
alleles in a wide variety of genetic backgrounds.
Table 2. Crucifer oilseed species/accessions tested for the presence of the
Fad3A allele using primers A048 and A050.
Species ' Type Accession 'Linolenic acid content
B. juncea Spring/breedingJ90-2741 High
B. juncea Spring/breedingJ90-4253 High
B. juncea Spring/breedingJ90-223 High
B. juncea Spring/breedingT097-3422-1 High
B. juncea Spring/breedingT097-3421-1 High
B. juncea Spring/breedingT097-3414 Low
B. juncea Spring/breedingT097-3400 High
B. napus Spring/breedingDH13830 High
B. napus Spring/breedingDH13619 High
B. napus Spring/breeding9592 High
B. napus Spring/canola Range High
B. napus Spring/canola Dunkeld High
B. napus Spring/breedingN89-17 High
B. napus Spring/breedingYN90-1016 High
B. napus Spring/breeding264-663 High
B. napus Spring/breeding1269 High
B. napus Spring/breeding1526 High
B. napus Spring/breedingS86-69 Low
B. rapa Spring/canola Horizon High
B. rapa Spring/canola Mavrick High
B. rapa Spring/canola Reward High
B. rapa Spring/canola Tobin High
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B. rapa Spring/rape Bronowski High
B. rapa Spring/rape Cresor High
B. rapa Spring/rape Midas High
B. rapa Spring/rape Oro High
B. napus Spring/canola AC Elect High
B. napus Spring/canola AC Excel High
B. napus Spring/canola AC H102 High
B. napus Spring/canola Alto High
B. napus Spring/canola Cyclone High
B. napus Spring/canola Delta High
B. napus Spring/canola Garrison High
B. napus Spring/canola Global High
B. napus Spring/canola Hyola 417 High
B. napus Spring/canola Karat High
B. napus Spring/canola Legacy High
B. napus Spring/canola Legend High
B. napus Spring/canola Polo High
B. napus Spring/canola Profit High
B. napus Spring/canola Regent High
B. napus Spring/canola Shiralee High
B. napus Spring/canola Stellar Low
B. napus Spring/canola Topas High
B. napus Spring/canola Tower High
B. napus Spring/canola Tribute High
B. napus Spring/canola Westar High
B. napus Winter/canola Cascade High
B. napus Winter/canola Ceres High
B. napus Winter/canola Glacier High
B. napus Winter/canola Mar High
B. napus Winter/canola Rubin High
B. napus Winter/canola Samourai High
B. napus Winter/canola Tandem High
B. napus Winter/canola Tapidor High
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B. napus Winter/rapeMarcus High
B. napus Winter/rapeJet Neuf High
B. juncea oriental AC Vulcan High
B. juncea oriental Forge High
B. juncea Brown Scimitar High
S. alba Spring/canolaWD96-2-3 High
S. alba Mustard Emergo High
B. rapa Spring/breeding7001 High
B. rapa Spring/breeding6909 High
B. rapa Spring/breeding6810 High
B. rapa Spring/breeding6794 High
I Winter and Spring
represent the
growth habit;
canola indicates
low in erucic
acid
and low in glucosinolate
content, rape
indicates high
erucic acid content,
breeding indicates
unregistered lines.
2Low = <4% >8% C 18:3.
C 18:3, High
=
Example 2
Figure 9 shows a protein sequence alignment between the Apollo Fad3A protein
and a wide variety of other Fad3 sequences, identified by database accession
number, and
more particularly described below. The alignment was produced using the BLASTP
software available from the National Centre for Biotechnology Information
(NCBI,
Bethesda, Maryland, U.S.A.) through the Internet at
http://www.cnbi.nlm.nih.gov/BLAST/.
A description of how to use this software, including how to optimally align
sequences is
available on the Internet at http://www.cnbi.nlm.nih.gov/BLAST/blast
help.html. In
summary form, the database sequences are as follows, with the 'Expect' value
of the match
with the Apollo Fad3A sequence, as calculated by the BLAST algorithm:
Table 3: Fad3 Sequences Compared2 to Apollo Fad3
Accession
3O spIP463111FD31BRANA OMEGA-3FATTYACIDDESATURASE,ENDOPLA.....Ø0
spIP486241FD32 BRANA OMEGA-3FATTYACIDDESATURASE,ENDOPLA.Ø0
spIP486231FD3E ARATH OMEGA-3FATTYACIDDESATURASE,ENDOPLA. 0.0
gi13133289 (AF020204) omega-3desaturase [Pelargonium .e-171
x hor.
spIP322911FD3E PHAAU OMEGA-3FATTYACIDDESATURASE,ENDOPLA..e-168
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gi14091113 (AF047172) omega-3 fatty acid desaturasee-168
[Vernic...
spIP486221FD3D ARATH TEMPERATURE-SENSITIVE OMEGA-3e-167
FATTY AC...
gbIAAD157441 (AF047039) omega-3 fatty acid desaturasee-166
[Peri...
spIP486191FD3C_RICCO OMEGA-3 FATTY ACID DESATURASE,e-165
CHLOROP...
S gi11754795 (U59477) omega-3 fatty acid desaturase e-164
[Perilla ...
spIP986201FD3C SESIN OMEGA-3 FATTY ACID DESATURASE,e-164
CHLOROP...
spIP963101FD3C ARATH OMEGA-3 FATTY ACID DESATURASE,e-164
CHLOROP...
dbjIBAAl14751 (D79979) omega-3 fatty acid desaturasee-163
[Nicot...
spIP48626~FD3E_TOBAC OMEGA-3 FATTY ACID DESATURASE,e-163
ENDOPLA...
gi14240385 (AF061027) omega-3 fatty acid desaturasee-162
precurs...
gi11786066 (U75745) omega-3 fatty acid desaturase e-162
[Petrosel...
spIP486251FD3E SOYBN OMEGA-3 FATTY ACID DESATURASE,e-162
ENDOPLA...
spIP486181FD3C BRANA OMEGA-3 FATTY ACID DESATURASE,e-162
CHLOROP...
dbjIBAA224401 (D63953) fatty acid desaturase [Zea e-162
mays] >gi...
IS sp~P486211FD3C SOYBN OMEGA-3 FATTY ACID DESATURASE,e-161
CHLOROP...
dbjIBAA224411 (D63954) fatty acid desaturase [Zea e-160
mays]
emb~CAA076381 (AJ007739) w-3 desaturase [Solanum e-160
tuberosum]
gi1699390 (U17063) delta-15 lineoyl desaturase e-155
[Limnanthes ...
dbjIBAA07785.11 (D43688) plastid omega-3 fatty e-154
acid desatur...
dbjIBAA283581 (D84678) omega-3 fatty acid desaturasee-154
[Triti...
dbjIBAA113971 (D78506) w-3 fatty acid desaturase e-147
[Oryza sat...
gi1408490 (L22963) omega-3 fatty acid desaturase e-145
[Brassica ...
dbjIBAA224391 (D63952) fatty acid desaturase [Zea e-113
mays]
dbjIBAAl13961 (D78505) w-3 fatty acid desaturase e-110
[Oryza sat...
2S gi12197199 (U36389) omega-3 desaturase [Synechococcuse-102
PCC7002]
gbIAAD41582.11AF056572 1 (AF056572) unknown [Brassicae-102
rapa]...
pirIIS52650 desaturase delta 15 - Synechocystis 6e-96
sp. (strain...
gbIAAD41581.11AF056571 1 (AF056571) unknown [Brassica6e-80
olera...
gbIAAD41580.1~AF0565701 (AF056570) unknown [Brassica2e-79
napus]
' Some "E" values shown as exponents, e.g. 'e-171 = 1x10-"'
The database used a basis for the BLASTP search was Non-redundant GenBank
CDS (translations+PDB+SwissProt+SPupdate+PIR), Posted date: Sep 14, 1999 3:12
PM
(number of letters in database: 126,047,814; number of sequences in database:
411,698),
using the following parameters:
3S Lambda K H
0.324 0.140 0.461
Gapped
Lambda K H
0.270 0.0470 0.230
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Matrix: BLOSUM62
Gap Penalties: Existence: 1 l, Extension: 1
Number of Hits to DB: 106686529
Number of Sequences: 411698
Number of extensions: 4746913
Number of successful extensions: 13626
Number of sequences better than 10.0: 129
Number of HSP's better than 10.0 without gapping: 102
Number of HSP's successfully gapped in prelim test: 27
Number of HSP's that attempted gapping in prelim test: 13347
Number of HSP's gapped (non-prelim): 139
length of query: 380
length of database: 126,047,814
effective HSP length: 48
effective length of query: 332
effective length of database: 106286310
effective search space: 35287054920
effective search space used: 35287054920
T: 11
A: 40
X1: 15 ( 7.0 bits)
X2: 3 8 ( 14. 8 bits)
X3: 64 (24.9 bits)
S 1: 40 (21.5 bits)
S2: 71 (32.1 bits)
Further particulars of the non-Apollo Fad3 sequences included in Figure 9 are
as
follows:
P46311 (Brassica nanus)
LOCUS FD31 BRANA 377 as PLN O1-FEB-1996
DEFINITION OMEGA-3 FATTY ACID DESATURASE, ENDOPLASMIC RETICULUM
VERSION 1).
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ACCESSION P46311
PID g1169600
VERSION P46311 GI:1169600
DBSOURCE swissprot: locus FD31 BRANA, accession P46311;
S class: standard.
created: Nov 1, 1995.
sequence updated: Nov 1, 1995.
annotation updated: Feb 1, 1996.
xrefs: gi: 408491, gi: 408492
1O KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; ENDOPLASMIC
RETICULUM;
TRANSMEMBRANE.
SOURCE rape.
ORGANISM Brassica napus
1S Eukaryotae; Viridiplantae; Charophyta/Embryophyta
group;
Embryophyta; Tracheophyta; seed plants; Magnoliophyta;
eudicotyledons; Rosidae; Capparales; Brassicaceae;
Brassica.
REFERENCE 1 (residues 1 to 377)
AUTHORS YADAV,N.S., WIERZBICKI,A., AEGERTER,M., CASTER,C.S.,
PEREZ-
2O GRAU,L.,KINNEY,A.J., HITZ,W.D., BOOTH,J.R. JR., SCHWEIGER,B.,
STECCA,K.L., ALLEN,S.M., BLACKWELL,M.,
REITER,R.S.,CARLSON,T.J., RUSSELL,S.H., FELDMANN,K.A.,
PIERCE, J. and BROWSE, J.
TITLE Cloning of higher plant omega-3 fatty acid desaturases
2S JOURNAL Plant Physiol. 103 (2), 467-476 (1993)
MEDLINE 94302147
REMARK SEQUENCE FROM N.A.
TISSUE=SEED
COMMENT [FUNCTION] ER (MICROSOMAL) OMEGA-3 FATTY ACID DESATURASE
3O INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS
OF 18:3
FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT MEMBRANES. IT IS
THOUGHT TO USE CYTOCHROME B5 AS AN ELECTRON DONOR AND TO ACT
ON FATTY ACIDS ESTERIFIED TO PHOSPHATIDYLCHOLINE AND,
POSSIBLY, OTHER PHOSPHOLIPIDS.
3S [PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
[SUBCELLULAR LOCATION] ENDOPLASMIC RETICULUM. [DOMAIN] THE
HISTIDINE BOX DOMAINS MAY CONTAIN THE ACTIVE SITE AND/OR BE
INVOLVED IN METAL ION BINDING.
[SIMILARITY] TO OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
4O FEATURES Location/Qualifiers
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source 1..377
/organism="Brassica napus"
/db xref="taxon:3708"
1..377
$ Protein 1..377
/product="OMEGA-3 FATTY ACID DESATURASE,
ENDOPLASMIC
RETICULUM"
/EC number="1.14.99.-"
Region 54..73
1~ /region name="Transmembrane region"
Region 92..96
/note="HISTIDINE BOX 1."
/region name="Domain"
Region 128..132
IS /note="HISTIDINE BOX 2."
/region name="Domain"
Region 203..226
/region name="Transmembrane region"
Region 233..251
2~ /region name="Transmembrane region"
Region 295..299
/note="HISTIDINE BOX 3."
/region name="Domain"
ORIGIN (SEQID N0: 9)
2$ mvvamdqrsn angderfdps aqppfkigdi raaipkhcwv ardifavval
ksplrsmsyv
avaavyfdsw ffwplywaaq gtlfwaifvl ghdcghgsfs hilhsfilvp
dipllntavg
yhgwrishrt hhqnhghven deswvplpek lyknlshstr layplylwyr
mlrytvplpm
spgkegshyn pysslfapse rkliatsttc wsimlatlvy lkvygvpyii
lsflvgpvtv
fvmwldavty lhhhghddkl pwyrgkewsy lrgglttidr digthvihhl
dygifnnihh
~ fpqiphyhlv datksakhvl gryyrepkts gaipihlves vsdtgdivfy
lvasikkdhy
etdpdlyvya sdkskin
P48624 (Brassica
napus)
LOCUS FD32 BRAVA 383 as PLN Ol-FEB-1996
3S DEFINITION OMEGA-3 FATTY ACID DESATURASE, ENDOPLASMICRETICULUM
VERSION 2).
ACCESSION P48624
PID 81345967
VERSION P48624 GI:1345967
~ DBSOURCE swissprot: locus FD32 BRAVA, accession
P48624;
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class: standard.
created: Feb 1, 1996.
sequence updated: Feb 1, 1996.
annotation updated: Feb 1, 1996.
xrefs: gi: 167147, gi: 167148
KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; ENDOPLASMIC
RETICULUM;
TRANSMEMBRANE.
SOURCE rape.
1~ ORGANISM Brassica napus
Eukaryotae; Viridiplantae; Charophyta/Embryophyta
group;
Embryophyta; Tracheophyta; seed plants; Magnoliophyta;
eudicotyledons; Rosidae; Capparales; Brassicaceae;
Brassica.
REFERENCE 1 (residues 1 to 383)
IS AUTHORS Arondel,V., Lemieux,B., Hwang,I., Gibson,S., Goodman,H.M.
and
Somerville,C.R.
TITLE Map-based cloning of a gene controlling omega-3 fatty
acid
desaturation in Arabidopsis
JOURNAL Science 258 (5086), 1353-1355 (1992)
2~ MEDLINE 93088059
REMARK SEQUENCE FROM N.A.
COMMENT [FUNCTION] ER (MICROSOMAL) OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS
OF 18:3
FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT MEMBRANES.
IT IS
2S THOUGHT TO USE CYTOCHROME B5 AS AN ELECTRON DONOR
AND TO ACT
ON FATTY ACIDS ESTERIFIED TO PHOSPHATIDYLCHOLINE
AND,
POSSIBLY, OTHER PHOSPHOLIPIDS.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
[SUBCELLULAR LOCATION] ENDOPLASMIC RETICULUM. [DOMAIN]
THE
3O HISTIDINE BOX DOMAINS MAY CONTAIN THE ACTIVE SITE
AND/OR BE INVOLVED IN METAL ION BINDING.
[SIMILARITY] TO OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
FEATURES Location/Qualifiers
source 1..383
3$ /organism="Brassica napus"
/db xref="taxon:3708"
1..383
Protein 1..383
/product="OMEGA-3 FATTY ACID DESATURASE, ENDOPLASMIC
4O RETICULUM"
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/EC number="1.14.99.-"
Region 53..73
/region name="Transmembrane "
region
Region 98..102
/note="HISTIDINE "
BOX l.
/region name="Domain"
Region 134..138
/note="HISTIDINE "
BOX 2.
/region name="Domain"
1~ Region 210..230
/region name="Transmembrane region"
Region 234..254
/region name="Transmembrane region"
Region 301..305
I$ /note="HISTIDINE "
BOX 3.
/region name="Domain"
ORIGIN (SEQ ID NO: )
mvvamdqrsn vngdsgarkeegfdpsaqpp fkigdiraaipkhcwvksplrsmsyvtrdi
favaalamaa vyfdswflwplywvaqgtlf waifvlghdcghgsfsdipllnsvvghilh
2~ sfilvpyhgw rishrthhqnhghvendesw vplpeklyknlphstrmlrytvplpmlayp
iylwyrspgk egshfnpysslfapserkli atsttcwsimlatlvylsflodpvtvlkvy
gvpyiifvmw ldavtylhhhghdeklpwyr gkewsylrgglttidrdygifnnihhdigt
hvihhlfpqi phyhlvdatraakhvlgryy repktsgaipihlveslvasikkdhyvsdt
gdivfyetdp dlyvyasdkskin
P48623 (thale
cress,
Arabidopsis
thaliana)
Score = bits (1922), Expect = 0.0
753
Identities 348/386 90%), Positives = 362/386(93s),Gaps = 6/386(10)
=
LOCUS FD3E ARATH 386 as PLN O1-OCT-1996
3O DEFINITION OMEGA-3 FATTY ACID DESATURASE, ENDOPLASMIC RETICULUM.
ACCESSION P48623
PID 81345973
VERSION P48623 GI:1345973
DBSOURCE swissprot: locus FD3E ARATH, accession P48623;
class: standard.
created: Feb 1, 1996.
sequence updated: Feb 1, 1996.
annotation updated: Oct l, 1996.
xrefs: 8i: 408482, 8i: 408483, 8i: 1030693, 8i:
471091, 8i:
511907, 8i: 1197795
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KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; ENDOPLASMIC
RETICULUM;
TRANSMEMBRANE.
SOURCE thale cress.
S ORGANISM Arabidopsis thaliana
Eukaryotae; Viridiplantae; Charophyta/Embryophyta
group;
Embryophyta; Tracheophyta; seed plants; Magnoliophyta;
eudicotyledons; Rosidae; Capparales; Brassicaceae;
Arabidopsis.
REFERENCE 1 (residues 1 to 386)
AUTHORS YADAV,N.S., WIERZBICKI,A., AEGERTER,M., CASTER,C.S.,
PEREZ-
GRAU,L., KINNEY,A.J., HITZ,W.D., BOOTH,J.R. JR.,
SCHWEIGER,B., STECCA,K.L., ALLEN,S.M., BLACKWELL,M.,
REITER,R.S., CARLSON,T.J., RUSSELL,S.H., FELDMANN,K.A.,
1S PIERCE, J. and BROWSE, J.
TITLE Cloning of higher plant omega-3 fatty acid desaturases
JOURNAL Plant Physiol. 103 (2), 467-476 (1993)
MEDLINE 94302147
REMARK SEQUENCE FROM N.A.
2O STRAIN=CV. COLUMBIA; TISSUE=SEEDLING
REFERENCE 2 (residues 1 to 386)
AUTHORS WATAHIKI,M.C. and YAMAMOTO,K.T.
TITLE Direct Submission
JOURNAL Submitted (??-SEP-1993) TO EMBL/GENBANK/DDBJ DATA
BANKS
ZS REMARK SEQUENCE FROM N.A.
STRAIN=CV. COLUMBIA; TISSUE=HYPOCOTYL
REFERENCE 3 (residues 1 to 386)
AUTHORS Nishiuchi,T., Nishimura,M., Arondel,V. and Iba,K.
TITLE Genomic nucleotide sequence of a gene encoding a
microsomal
30 omega-3 fatty acid desaturase from Arabidopsis thaliana
JOURNAL Plant Physiol. 105 (2), 767-768 (1994)
MEDLINE 94345020
REMARK SEQUENCE FROM N.A.
STRAIN=CV. COLUMBIA
3S COMMENT [FUNCTION] MICROSOMAL (ER) OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS
OF 18:3
FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT MEMBRANES.
IT IS
THOUGHT TO USE CYTOCHROME B5 AS AN ELECTRON DONOR
AND TO ACT
ON FATTY ACIDS ESTERIFIED TO PHOSPHATIDYLCHOLINE
AND,
4O POSSIBLY, OTHER PHOSPHOLIPIDS.
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[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
[SUBCELLULAR LOCATION] ENDOPLASMIC RETICULUM.
[TISSUE SPECIFICITY] ABUNDANT IN LEAVES AND SEEDLINGS.
BARELY
DETECTABLE IN ROOT TISSUE. [DOMAIN] THE HISTIDINE
BOX DOMAINS
S MAY CONTAIN THE ACTIVE SITE AND/OR BE INVOLVED IN
METAL ION
BINDING.
[SIMILARITY] TO OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
FEATURES Location/Qualifiers
source 1..386
/organism="Arabidopsis thaliana"
/db xref="taxon:3702"
1..386
Protei n 1..386
/product="OMEGA-3 FATTY ACID DESATURASE, ENDOPLASMIC
IS RETICULUM"
/EC number="1.14.99.-"
Region 63..83
/region name="Transmembrane region"
Region 101..105
/note="HISTIDINE BOX 1."
/region name="Domain"
Region 137..141
/note="HISTIDINE BOX 2."
/region name="Domain"
2S Region 220..240
/region name="Transmembrane region"
Region 242..262
/region name="Transmembrane region"
Region 304..308
/note="HISTIDINE BOX 3."
/region name="Domain"
ORIGIN (SEQID NO: 11)
mvvamdqrtn vngdpgagdr kkeerfdpsa qppfkigdir aaipkhcwvk splrsmsyvv
rdiiavaala iaavyvdswf lwplywaaqg tlfwaifvlg hdcghgsfsd ipllnsvvgh
3S ilhsfilvpy hgwrishrth hqnhghvend eswvplperv ykklphstrm lrytvplpml
ayplylcyrs pgkegshfnp ysslfapser kliatsttcw simfvslial sfvfgplavl
kvygvpyiif vmwldavtyl hhhghdeklp wyrgkewsyl rgglttidrd ygifnnihhd
igthvihhlf pqiphyhlvd atkaakhvlg ryyrepktsg aipihlvesl vasikkdhyv
sdtgdivfye tdpdlyvyas dkskin
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3133289
(Pelar~onium
x hortorum)
LOCUS AAC16443 407 as PLN 15-MAY-1998
DEFINITION omega-3 desaturase.
ACCESSION AAC16443
S PID 83133289
VERSION AAC16443.1 GI:3133289
DBSOURCE accession AF020204.1
KEYWORDS
SOURCE Pelargonium x hortorum.
1~ ORGANISM Pelargonium x hortorum
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Rosidae; Geraniales;
Geraniaceae; Pelargonium.
1$ REFERENCE 1 (residues 1 to 407)
AUTHORS Schultz,D.J., Mumma,R.O., Cox-Foster,D., Craig,R.
and
Medford,J.I.
TITLE Geranium omega-3 desaturase
JOURNAL Unpublished
2~ REFERENCE 2 (residues 1 to 407)
AUTHORS Schultz,D.J., Mumma,R.O., Cox-Foster,D., Craig,R.
and
Medford,J.I.
TITLE Direct Submission
JOURNAL' Submitted (19-AUG-1997) Botany, MSU, 166 Plant Biology
25 Building, East Lansing, MI 48824, USA
COMMENT Method: conceptual translation supplied by author.
FEATURES Location/Qualifiers
source 1..407
/organism="Pelargonium x hortorum"
3~ /db xref="taxon:4031"
Protei n <1..407
/product="omega-3 desaturase"
CDS 1..407
/gene="pxh-15"
35 /coded by="AF020204.1:<1..1226"
ORIGIN (SEQID N0: 12)
sdfdp sapppfrlge iraaipqhcw vkspwrsmsy vvrdivvvfa lavaafrlds
wlvwpiywav
qgtmfwaifv lghdcghgsf sdshilnsvm ghilhssilv pyhgwrishk thhsnhghve
ndeswvplte ktyksldvst rllrftipfp vfaypfylww rspgkkgshf npysdlfaps
~ errdvltsti swsimvalla glscvfglvp mlklyggpyw ifvmwldtvt ylhhhghddh
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klpwyrgkew sylrgglttv drdyglfnni hhdigthvih hlfpqiphyh lveatraakp
vlgkyyrepk rsgpfpyhli dnlvksiked hyvsdtgdiv fyetdpeqfk sdpkkl
P32291 (mun~
bean, Vilna
radiata)
S Score = bits (1507), Expect = e-168
591
Identities = 259/359 (72%), Positives = 303/359 (84%)
LOCUS FD3E PHAAU 380 as PLN O1-FEB-1996
DEFINITION OMEGA-3 FATTY ACID DESATURASE, ENDOPLASMIC RETICULUM
IO (INDOLE-3-ACETIC ACID INDUCED PROTEIN ARG1).
ACCESSION P32291
PID 8416638
VERSION P32291 GI:416638
DBSOURCE swissprot: locus FD3E-PHAAU, accession P32291;
IS class: standard.
created: Oct l, 1993.
sequence updated: Oct 1, 1993.
annotation updated: Feb 1, 1996.
xrefs: 8i: 287561, 8i: 287562
ZO KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; ENDOPLASMIC
RETICULUM; TRANSMEMBRANE.
SOURCE mung bean.
ORGANISM Vigna radiata
Eukaryotae; Viridiplantae; Charophyta/Embryophyta
group;
2S Embryophyta; Tracheophyta; seed plants; Magnoliophyta;
eudicotyledons; Rosidae; Fabales; Fabaceae; Papilionoideae;
Vigna.
REFERENCE 1 (residues 1 to 380)
AUTHORS YAMAMOTO,K.T., MORI,H. and IMASEKI,H.
3O JOURNAL PLANT CELL PHYSIOL. 33, 13-20 (1992)
REMARK SEQUENCE FROM N.A.
TISSUE=HYPOCOTYL
COMMENT [FUNCTION] MICROSOMAL (ER) OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS
OF 18:3
3S FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT MEMBRANES.
IT IS
THOUGHT TO USE CYTOCHROME B5 AS AN ELECTRON DONOR
AND TO ACT
ON FATTY ACIDS ESTERIFIED TO PHOSPHATIDYLCHOLINE
AND,
POSSIBLY, OTHER PHOSPHOLIPIDS.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
4O [SUBCELLULAR LOCATION] ENDOPLASMIC RETICULUM. INDUCTION]
BY
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AUXIN, ETHYLENE AND WOUNDING. [DOMAIN] THE HISTIDINE BOX
DOMAINS MAY CONTAIN THE ACTIVE SITE AND/OR BE INVOLVED IN
METAL ION BINDING. [SIMILARITY] TO OTHER PLANT OMEGA-3 FATTY
ACID DESATURASES.
FEATURES Location/Qualifiers
source 1..380
/organism="Vigna radiata"
/db xref="taxon:3916"
1..380
Protein 1..380
/product="OMEGA-3 FATTY ACID DESATURASE, ENDOPLASMIC
RETICULUM"
/EC number="1.14.99.-"
Region 59..78
/region name="Transmembrane region"
Region 97..101
/note="HISTIDINE BOX 1."
/region name="Domain"
Region 133..137
/note="HISTIDINE BOX 2."
/region name="Domain"
Region 208..231
/region name="Transmembrane region"
Region 238..256
/region name="Transmembrane region"
Region 300..304
/note="HISTIDINE BOX 3."
/region name="Domain"
ORIGIN (SEQ ID N0: 13)
fdpgapppf kiadiraaip khcwekstlr slsyvlrdvl vvtalaasai sfnswffwpl
ywpaqgtmfw alfvlghdcg hgsfsnsskl nsfvghilhs lilvpyngwr ishrthhqnh
ghvekdeswv pltekvyknl ddmtrmlrys fpfpifaypf ylwnrspgke gshfnpysnl
fspgerkgvv tstlcwgivl svllylslti gpifmlklyg vpylifvmwl dfvtylhhhg
ythklpwyrg qewsylrggl ttvdrdygwi nnvhhdigth vihhlfpqip hyhlveatks
3$ aksvlgkyyr epqksgplpf hllkyllqsi sqdhfvsdtg divyyqtdpk lhqdswtksk
4091113 (Yernicia fordiil
Score = 590 bits (1504), Expect = e-168
Identities = 265/377 (70~), Positives = 305/377 (80~), Gaps = 7/377 (1s)
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LOCUS AAC98967 387 as PLN O1-JAN-1999
DEFINITION omega-3 fatty acid desaturase.
ACCESSION AAC98967
PID g4091113
VERSION AAC98967.1 GI:4091113
DBSOURCE locus AF047172 accession AF047172.1
KEYWORDS
SOURCE Vernicia fordii.
ORGANISM Vernicia fordii
1~ Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Rosidae; eurosids
I;
Malpighiales; Euphorbiaceae; Vernicia.
REFERENCE 1 (residues 1 to 387)
1$ AUTHORS Tang,F., Dyer,J.M., Lax,A.R., Shih,D.S., Chapital,D.C.
and
Pepperman,A.B.
TITLE Nucleotide sequence of a cDNA clone for endoplasmic
reticular
Fatty acid desaturase from Aleurites fordii seeds
JOURNAL Unpublished
2~ REFERENCE 2 (residues 1 to 387)
AUTHORS Tang, F.
TITLE Direct Submission
JOURNAL Submitted (06-FEB-1998) Southern Regional Research
Center,
USDA-ARS, 1100 Robert E. Lee Blvd., New Orleans,
LA 70179,
25 USA
COMMENT Method: conceptual translation supplied by author.
FEATURES Location/Qualifiers
source 1..387
/organism="Vernicia fordii"
3~ /variety="L-2"
/db xref="taxon:73154"
/dev stage="seed"
Protein 1..387
/product="omega-3 fatty acid desaturase"
35 CDS 1..387
/gene="Fad3"
/coded by="AF047172.1:39..1202"
ORIGIN (SEQ ID N0: 14)
ngvngfha keeeeeedfd lsnpppfnig qiraaipkhc wvknpwrslt yvfrdvvvvf
~ alaaaafyfn swlfwplywf aqgtmfwaif vlghdcghgs fsnnsslnnv vghllhssil
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vpyhgwrish rthhqnhgnv ekdeswvplp ekiykemdls trilrysvpl pmfalpfylw
wrspgkegsh fnpnsdffap herkavltsn fcfsimalll lyscfvfgpv qvlkfygipy
lvfvmwldfv tymhhhghee klpwyrgkew sylrgglqtv drdygwinni hhdigthvih
hlfpqiphyh lieatkaakp vlgkyyrepk ksgpfpfhlf snlvrsmsed hyvsdigdiv
S fyqtdpdiyk vdkskln
P48622
(Arabidopsis
thaliana)
LOCUS FD3D ARATH 435 as PLN O1-FEB-1996
DEFINITIONTEMPERATURE-SENSITIVE OMEGA-3 FATTY ACID DESATURASE,
IO CHLOROPLAST PRECURSOR.
ACCESSION P48622
PID g1345972
VERSION P48622 GI:1345972
DBSOURCE swissprot: locus FD3D ARATH, accession P48622;
1S class: standard.
created: Feb 1, 1996.
sequence updated: Feb 1, 1996.
annotation updated: Feb 1, 1996.
xrefs: gi: 516044, gi: 516045, gi: 497218, gi: 497219,
gi:
2O 1030694, gi: 471093
KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; CHLOROPLAST;
MEMBRANE; TRANSIT PEPTIDE.
SOURCE thale cress.
ORGANISM Arabidopsis thaliana Eukaryotae; Viridiplantae;
2S Charophyta/Embryophyta group; Embryophyta; Tracheophyta;
seed
plants; Magnoliophyta; eudicotyledons; Rosidae; Capparales;
Brassicaceae; Arabidopsis.
REFERENCE 1 (residues 1 to 435)
AUTHORS Gibson,S., Arondel,V., Iba,K. and Somerville,C.
3O TITLE Cloning of a temperature-regulated gene encoding
a
chloroplast omega-3 desaturase from Arabidopsis thaliana
JOURNAL Plant Physiol. 106 (4), 1615-1621 (1994)
MEDLINE 95148742
REMARK SEQUENCE FROM N.A.
3S STRAIN=CV. COLUMBIA; TISSUE=AERIAL PARTS
REFERENCE 2 (residues 1 to 435)
AUTHORS WATAHIKI,M.C. and YAMAMOTO,K.T.
TITLE Direct Submission
JOURNAL Submitted (??-SEP-1993) TO EMBL/GENBANK/DDBJ DATA
BANKS
4O REMARK SEQUENCE FROM N.A.
36
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STRAIN=CV. COLUMBIA; TISSUE=HYPOCOTYL
COMMENT [FUNCTION] CHLOROPLAST OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS OF 16:3
AND 18:3 FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT
S MEMBRANES. IT IS THOUGHT TO USE FERREDOXIN AS AN ELECTRON
DONOR AND TO ACT ON FATTY ACIDS ESTERIFIED TO GALACTOLIPIDS,
SULFOLIPIDS AND PHOSPHATIDYLGLYCEROL.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
[SUBCELLULAR LOCATION] CHLOROPLAST, MEMBRANE-BOUND
IO (PROBABLE). [INDUCTION] BY LOW TEMPERATURES. [DOMAIN] THE
HISTIDINE BOX DOMAINS MAY CONTAIN THE ACTIVE SITE
AND/OR BE INVOLVED IN METAL ION BINDING. [SIMILARITY] TO
OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
FEATURES Location/Qualifiers
IS source 1..435
/organism="Arabidopsis thaliana"
/db xref="taxon:3702"
1..435
Protein /product="TEMPERATURE-1..435
ZO SENSITIVE OMEGA-3 FATTY ACID
DESATURASE, CHLOROPLAST PRECURSOR"
/EC number="1.14.99.-"
Region 1..(2.435)
/region name="Transit peptide"
ZS /note="CHLOROPLAST."
Region (1.434)..435
/region name="Mature chain"
/note="TEMPERATURE-SENSITIVE OMEGA-3 FATTY ACID
DESATURASE, CHLOROPLAST."
30 Region 156..160
/region name="Domain"
/note="HISTIDINE BOX 1."
Region 192..196
/region name="Domain"
3S /note="HISTIDINE BOX 2."
Region 359..363
/region name="Domain"
/note="HISTIDINE BOX 3."
ORIGIN (SEQ ID NO: 15)
40 r fdpgapppfn ladiraaipk hcwvknpwms msyvvrdvai vfglaavaay fnnwllwply
37
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wfaqgtmfwa lfvlghdcgh gsfsndprln svaghllhss ilvpyhgwri shrthhqnhg
hvendeswhp lpesiyknle kttqmfrftl pfpmlaypfy lwnrspgkqg shyhpdsdlf
lpkekkdvlt stacwtamaa llvclnfvmg piqmlklygi pywifvmwld fvtylhhhgh
edklpwyrgk ewsylrgglt tldrdygwin nihhdigthv ihhlfpqiph yhlveateaa
kpvlgkyyre pknsgplplh llgsliksmk qdhfvsdtgd vvyyeadpkl
AAD15744 (Perilla frutescens)
LOCUS AAD15744 391 as PLN 03-MAR-1999
DEFINITION omega-3 fatty acid desaturase.
l~ ACCESSION AAD15744
PID 84321399
VERSION AAD15744.1 GI:4321399
DBSOURCE locus AF047039 accession AF047039.1
KEYWORDS
IS SOURCE Perilla frutescens.
ORGANISM Perilla frutescens
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Asteridae; euasterids
I;
Lamiales; Lamiaceae; Perilla.
REFERENCE 1 (residues 1 to 391)
AUTHORS Chung,C.-H., Kim,J.-L., Lee,Y.-C. and Choi,Y.-L.
TITLE Molecular cloning and characterization of a omega-3
cDNA from
perilla seed
25 JOURNAL Unpublished
REFERENCE 2 (residues 1 to 391)
AUTHORS Chung,C.-H., Kim,J.-L., Lee,Y.-C. and Choi,Y.-L.
TITLE Direct Submission
JOURNAL Submitted (07-FEB-1998) Biotechnology, Dong-A University,
30 840, Ha-Dan-Dong, Sa-Ha-Gu, Pusan 604-714, South
Korea
COMMENT Method: conceptual translation.
FEATURES Location/Qualifiers
source 1..391
/organism="Perilla frutescens"
35 /cultivar="Suwon-8"
/db xref="taxon:48386"
/dev stage="seed"
Protein 1..391
/product="omega-3 fatty acid desaturase"
40 CDS 1..391
38
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/gene="FAD3"
/coded by="AF047039.1:156..1331"
ORIGIN (SEQ ID N0:16)
$ gk raadkfdpaa pppfkiadir aaipahcwvk npwrslsyvv wdvaavfall aaavyinswa
fwpvywiaqg tmfwalfvlg hdcghgsfsd nttlnnvvgh vlhssilvpy hgwrishrth
hqnhghvekd eswvplpenl ykkldfstkf lrykipfpmf ayplylwyrs pgktgshfnp
ysdlfkpner glivtstmcw aamgvfllya stivgpnmmf klygvpylif vmwldtvtyl
hhhgydkklp wyrskewsyl rgglttvdqd ygffnkihhd igthvihhlf pqiphyhlve
1~ atreakrvlg nyyreprksg pvplhlipal lkslgrdhyv sdngdivyyq tddelf I
P48619 (Ricinus communis)
LOCUS FD3C RICCO 460 as PLN 15-DEC-1998
DEFINITION OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST PRECURSOR.
1$ ACCESSION P48619
PID 81345969
VERSION P48619 GI:1345969
DBSOURCE swissprot: locus FD3C RICCO, accession P48619;
class: standard.
20 created: Feb 1, 1996.
sequence updated: Feb 1, 1996.
annotation updated: Dec 15, 1998.
xrefs: 8i: 414731, 8i: 414732
xrefs (non-sequence databases): PFAM PF00487
ZS KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; CHLOROPLAST;
MEMBRANE; TRANSIT PEPTIDE.
SOURCE castor bean.
ORGANISM Ricinus communis
Eukaryota; Viridiplantae; Charophyta/Embryophyta group;
3~ Embryophyta; Tracheophyta; euphyllophytes; Spermatophyta;
Magnoliophyta; eudicotyledons; Rosidae; Euphorbiales;
Euphorbiaceae; Ricinus.
REFERENCE 1 (residues 1 to 460)
AUTHORS van de Loo,F.J. and Somerville,C.
35 TITLE Plasmid omega-3 fatty acid desaturase cDNA from Ricinus
communis
JOURNAL Plant Physiol. 105 (1), 443-444 (1994)
MEDLINE 94302177
REMARK SEQUENCE FROM N.A.
4O STRAIN=CV. BAKER 296; TISSUE=SEED
39
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[FUNCTION] CHLOROPLAST OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS OF 16:3
AND 18:3 FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT
MEMBRANES. IT IS THOUGHT TO USE FERREDOXIN AS AN ELECTRON
S DONOR AND TO ACT ON FATTY ACIDS ESTERIFIED TO GALACTOLIPIDS,
SULFOLIPIDS AND PHOSPHATIDYLGLYCEROL.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
[SUBCELLULAR LOCATION] CHLOROPLAST, MEMBRANE-BOUND
(PROBABLE). [DOMAIN] THE HISTIDINE BOX DOMAINS MAY CONTAIN
IO THE ACTIVE SITE AND/OR BE INVOLVED IN METAL ION BINDING.
[SIMILARITY] TO OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
FEATURES Location/Qualifiers
source 1..460
/organism="Ricinus communis"
IS /db xref="taxon:3988"
1..460
Protein 1..460
/product="OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST
PRECURSOR"
20 /EC number="1.14.99.-"
Region 1..(2.460)
/note="CHLOROPLAST."
/region name="Transit peptide"
Region (1.459)..460
ZS /note="OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST."
/region name="Mature chain"
Region 177..181
/note="HISTIDINE BOX 1."
/region name="Domain"
30 Region 213..217
/note="HISTIDINE BOX 2."
/region name="Domain"
Region 380..384
/note="HISTIDINE BOX 3."
3S /region name="Domain"
ORIGIN (SEQ ID N0:17)
ereefng ivnvdegkge ffdagapppf tladiraaip khcwvknpwr smsyvlrdvv vvfglaavaa
yfnnwvawpl ywfcqgtmfw alfvlghdcg hgsfsnnpkl nsvvghllhs silvpyhgwr
ishrthhqnh ghvendeswh plsekifksl dnvtktlrfs lpfpmlaypf ylwsrspgkk
40 gshfhpdsgl fvpkerkdii tstacwtama allvylnfsm gpvqmlklyg ipywifvmwl
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dfvtylhhhg hedklpwyrg kawsylrggl ttldrdygwi nnihhdigth vihhlfpqip
hyhlveatea akpvmgkyyr epkksgplpl hllgslvrsm kedhyvsdtg dvvyyqkdpk
lsgiggekte
1754795 (Perilla f'rutescens)
LOCUS AAB39387 438 as PLN 28-DEC-1996
DEFINITION omega-3 fatty acid desaturase.
ACCESSION AAB39387
PID 81754795
VERSION AAB39387.1 GI:1754795
DBSOURCE locus PFU59977 accession U59477.1
KEY4dORDS
SOURCE Perilla frutescens.
ORGANISM Perilla frutescens
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Asteridae; euasterids I;
Lamiales; Lamiaceae; Perilla.
REFERENCE 1 (residues 1 to 438)
AUTHORS Lee,S.-K., Kim,K.-H., Kim,Y.-M. and Hwang,Y.-S.
TITLE Cloning of plant omega-3 fatty acid desaturase gene from
Perilla frutescens
JOURNAL Unpublished
REFERENCE 2 (residues 1 to 438)
ZS AUTHORS Lee,S.-K.
TITLE Direct Submission
JOURNAL Submitted (30-MAY-1996) Biochemistry, National Agricultural
Science and Technology Institute, 249 Seodundong, Suwon 441-
707, Republic of Korea
FEATURES Location/Qualifiers
source 1..438
/organism="Perilla frutescens"
/strain="Okdong"
/db xref="taxon:48386"
/clone="Pfrfad7"
/dev stage="seedling"
Protein 1..438
/product="omega-3 fatty acid desaturase"
CDS 1..438
/coded by="U59477.1:222..1538"
41
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ORIGIN (SEQ ID N0: 18)
eergsv ivngvdefdp gapppfklsd iraaipkhcw vkdpwrsmsy vvrdvvvvfg laaaaayfnn
wavwpiywfa qstmfwalfv lghdcghgsf sndpklnsva ghllhssilv pyhgwrishr
thhqnhghve ndeswhpipe kiyrtldfat kklrftlpfp mlaypfylwg rspgkkgshf
hpdsdlfvpn erkdvitstv cwtamvaila glsfvmgpvq llklygipyi gfvawldlvt
ylhhhghdek lpwyrgkews ylrgglttld rdygwinnih hdigthvihh lfpqiphyhl
ieataaakpv lgkyykepkk sgpfpfyllg vlqksmkkdh yvsdtgdivy yqtdpe
P48620 (sesame, Sesamum indicum)
LOCUS FD3C SESIN 447 as PLN 15-DEC-1998
DEFINITION OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST PRECURSOR.
ACCESSION P48620
PID 81345970
VERSION P48620 GI:1345970
DBSOURCE swissprot: locus FD3C SESIN, accession P48620;
class: standard.
created: Feb 1, 1996.
sequence updated: Feb 1, 1996.
annotation updated: Dec 15, 1998.
xrefs: 8i: 870783, 8i: 870784
xrefs (non-sequence databases): PFAM PF00487
KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; CHLOROPLAST;
MEMBRANE; TRANSIT PEPTIDE.
SOURCE sesame.
ORGANISM Sesamum indicum
Eukaryota; Viridiplantae; Charophyta/Embryophyta
group;
Embryophyta; Tracheophyta; euphyllophytes; Spermatophyta;
Magnoliophyta; eudicotyledons; Asteridae; Gentiananae;
Lamiales; Pedaliaceae; Sesamum.
~ REFERENCE 1 (residues 1 to 447)
AUTHORS SHOJI, K.
TITLE Direct Submission
JOURNAL Submitted (??-APR-1995) TO EMBL/GENBANK/DDBJ DATA
BANKS
REMARK SEQUENCE FROM N.A.
3S STRAIN=CV. 4294; TISSUE=COTYLEDON
[FUNCTION] CHLOROPLAST OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS
OF 16:3
AND 18:3 FATTY ACIDS, IMPORTANT CONSTITUENTS OF
PLANT
MEMBRANES. IT IS THOUGHT TO USE FERREDOXIN AS AN
ELECTRON
4O DONOR AND TO ACT ON FATTY ACIDS ESTERIFIED TO GALACTOLIPIDS,
42
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SULFOLIPIDS AND PHOSPHATIDYLGLYCEROL.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
[SUBCELLULAR LOCATION] CHLOROPLAST, MEMBRANE-BOUND
(PROBABLE). [DOMAIN] THE HISTIDINE BOX DOMAINS MAY CONTAIN
S THE ACTIVE SITE AND/OR BE INVOLVED IN METAL ION BINDING.
[SIMILARITY] TO OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
FEATURES Location/Qualifiers
source 1..447
/organism="Sesamum indicum"
/db xref="taxon:4182"
1..447
Protein 1..447
/product="OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST
PRECURSOR"
IS /EC number="1.14.99.-"
Region 1..(2.447)
/note="CHLOROPLAST."
/region name="Transit peptide"
Region (1.446)..447
ZO /note="OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST."
/region name="Mature chain"
Region 167..171
/note="HISTIDINE BOX 1."
/region name="Domain"
25 Region 203..207
/note="HISTIDINE BOX 2."
/region name="Domain"
Region 370..374
/note="HISTIDINE BOX 3."
30 /region name="Domain"
ORIGIN (SEQ ID NO: 19)
a efdpgapppf klsdireaip khcwvkdpwr smgyvvrdva vvfglaavaa yfnnwvvwpl
ywfaqstmfw alfvlghdcg hgsfsndpkl nsvvghilhs silvpyhgwr ishrthhqnh
ghvendeswh plsekiyknl dtatkklrft lpfpllaypi ylwsrspgkq gshfhpdsdl
3$ fvpnekkdvi tstvcwtaml allvglsfvi gpvqllklyg ipylgnvmwl dlvtylhhhg
hedklpwyrg kewsylrggl ttldrdygwi nnihhdigth vihhlfpqip hyhlieatea
akpvlgkyyr epkksaplpf hllgdltrsl krdhyvsdvg dvvyyqtdpq 1
P46310 (Arabidopsis thaliana)
40 LOCUS FD3C ARATH 446 as PLN Ol-FEB-1996
43
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DEFINITION OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST PRECURSOR.
ACCESSION P46310
PID 81169599
VERSION P46310 GI:1169599
$ DBSOURCE swissprot: locus FD3C ARATH, accession P46310;
class: standard.
created: Nov 1, 1995.
sequence updated: Nov l, 1995.
annotation updated: Feb l, 1996.
1~ xrefs: 8i: 408480, 8i: 408481, 8i: 461160, 8i: 541653,
8i:
809491, 8i: 468434
KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; CHLOROPLAST;
MEMBRANE; TRANSIT PEPTIDE.
SOURCE thale cress.
1$ ORGANISM Chloroplast Arabidopsis thaliana
Eukaryotae; Viridiplantae; Charophyta/Embryophyta
group;
Embryophyta; Tracheophyta; seed plants; Magnoliophyta;
eudicotyledons; Rosidae; Capparales; Brassicaceae;
Arabidopsis.
20 REFERENCE 1 (residues 1 to 446)
AUTHORS YADAV,N.S., WIERZBICKI,A., AEGERTER,M., CASTER,C.S.,
PEREZ-
GRAU,L., KINNEY,A.J., HITZ,W.D., BOOTH,J.R. JR.,
SCHWEIGER,B., STECCA,K.L., ALLEN,S.M., BLACKWELL,M.,
REITER,R.S., CARLSON,T.J., RUSSELL,S.H., FELDMANN,K.A.,
25 PIERCE, J. and BROWSE, J.
TITLE Cloning of higher plant omega-3 fatty acid desaturases
JOURNAL Plant Physiol. 103 (2), 467-476 (1993)
MEDLINE 94302147
REMARK SEQUENCE FROM N.A.
3O STRAIN=CV. COLUMBIA; TISSUE=HYPOCOTYL
REFERENCE 2 (residues 1 to 446)
AUTHORS Iba,K., Gibson,S., Nishiuchi,T., Fuse, T., Nishimura,M.,
Arondel,V., Hugly,S. and Somerville,C.
TITLE A gene encoding a chloroplast omega-3 fatty acid
desaturase
35 complements alterations in fatty acid desaturation
and
chloroplast copy number of the fad? mutant of Arabidopsis
thaliana
JOURNAL J. Biol. Chem. 268 (32), 24099-24105 (1993)
MEDLINE 94043239
4O REMARK SEQUENCE FROM N.A.
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STRAIN=CV. COLUMBIA; TISSUE=AERIAL PARTS
REFERENCE 3 (residues 1 to 446)
AUTHORS WATAHIKI,M. and YAMAMOTO,K.
TITLE Direct Submission
S JOURNAL Submitted (??-NOV-1993) TO EMBL/GENBANK/DDBJ DATA BANKS
REMARK SEQUENCE FROM N.A.
STRAIN=CV. COLUMBIA; TISSUE=HYPOCOTYL
COMMENT [FUNCTION] CHLOROPLAST OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS OF 16:3
IO AND 18:3 FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT
MEMBRANES. IT IS THOUGHT TO USE FERREDOXIN AS AN ELECTRON
DONOR AND TO ACT ON FATTY ACIDS ESTERIFIED TO GALACTOLIPIDS,
SULFOLIPIDS AND PHOSPHATIDYLGLYCEROL.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
IS [SUBCELLULAR LOCATION] CHLOROPLAST, MEMBRANE-BOUND
(PROBABLE). [TISSUE SPECIFICITY] MOST ABUNDANT IN LEAVES AND
SEEDLINGS. [DOMAIN] THE HISTIDINE BOX DOMAINS MAY CONTAIN THE
ACTIVE SITE AND/OR BE INVOLVED IN METAL ION BINDING.
[SIMILARITY] TO OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
20 FEATURES Location/Qualifiers
source 1..446
/organism="Arabidopsis thaliana"
/chloroplast
/db xref="taxon:3702"
2S 1..446
Protein 1..446
/product="OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST
PRECURSOR"
/EC number="1.14.99.-"
Region 1..(2.446)
/note="CHLOROPLAST."
/region name="Transit peptide"
Region (1.445)..446
/note="OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST."
3S /region name="Mature chain"
Region 163..167
/note="HISTIDINE BOX 1."
/region name="Domain"
Region 199..203
4~ /note="HISTIDINE BOX 2."
4S
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/region name="Domain"
Region 366..370
/note="HISTIDINE BOX 3."
/region name="Domain"
S ORIGIN (SEQ ID N0:20)
espl eednkqrfdp gapppfnlad iraaipkhcw vknpwkslsy vvrdvaivfa laagaaylnn
wivwplywla qgtmfwalfv lghdcghgsf sndpklnsvv ghllhssilv pyhgwrishr
thhqnhghve ndeswhpmse kiyntldkpt rffrftlplv mlaypfylwa rspgkkgshy
hpdsdlflpk erkdvltsta cwtamaallv clnftigpiq mlklygipyw invmwldfvt
1~ ylhhhghedk lpwyrgkews ylrgglttld rdyglinnih hdigthvihh lfpqiphyhl
veateaakpv lgkyyrepdk sgplplhlle ilaksikedh yvsdegevvy ykadpnly
BAA11475 (Nicotiana tabacum)
LOCUS BAA11475 441 as PLN 05-FEB-1999
1S DEFINITION omega-3 fatty acid desaturase.
ACCESSION BAA11475
PID g1694625
VERSION BAA11475.1 GI:1694625
DBSOURCE locus D79979 accession D79979.1
ZO KEYWORDS
SOURCE common tobacco.
ORGANISM Nicotiana tabacum
Eukaryota; Viridip~antae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
2S eudicotyledons; Asteridae; Solananae; Solanales;
Solanaceae;
Nicotiana.
REFERENCE 1 (residues 1 to 441)
AUTHORS Hamada,T.
TITLE Direct Submission
JOURNAL Submitted (12-DEC-1995) to the DDBJ/EMBL/GenBank
databases. Tatsurou Hamada, Faculty of Science, Kyushu
University, Department of Biology; 6-10-1 Hakozaki,
Higashi-
ku, Fukuoka, Fukuoka 812, Japan
(Te1:092-641-1101(ex.4414), Fax:092-632-2741)
3S REFERENCE 2 (residues 1 to 441)
AUTHORS Hamada,T.
JOURNAL Unpublished (1995)
REFERENCE 3 (residues 1 to 441)
AUTHORS Hamada,T., Nishiuchi,T., Kodama,H., Nishimura,M.
and Iba,K.
4~ TITLE cDNA cloning of a wounding-inducible gene encoding
a plastid
46
CA 02386111 2002-03-28
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omega-3 fatty acid desaturase from tobacco
JOURNAL Plant Cell Physiol. 37 (5), 606-611 (1996)
MEDLINE 96416425
FEATURES Location/Qualifiers
S source 1..441
/organism="Nicotiana tabacum"
/db xref="taxon:4097"
/clone="lambda H 1"
/clone lib="lambda gtll"
I~ Protei n 1..441
/product="omega-3 fatty acid desaturase"
CDS 1..441
/gene="NtFAD7"
/coded by="D79979.1:28..1353"
IS ORIGIN ID NO: 21)
(SEQ
eeesertn ggeffdpg apppfklsdi kaaipkhcwv knpwksmsyv vrdvaivfgl
ns
aaaaayfnnwvvwplywfaq stmfwalfvl ghdcghgsfs nnhklnsvvg hilhssilvp
yhgwrishrthhqnhghven deswhpipek iynsldlatk klrftlpfpl laypfylwsr
spgkkgshfdpnsdlfvpse kkdvmtstlc wtamaallvg lsfvmgpfqv lklygipywg
20 fvmwldlvtylhhhghddkl pwyrgeewsy lrgglttldr dygwinnihh digthvihhl
fpqiphyhlveateaakpvl gkyykepkks gplpfyllgv liksmkqdhy vsdtgdivyy
rtdpqlsgfqk
P48626
(Nicotiana
tabacum)
ZS LOCUS FD3E TOBAC 379 as PLN O1-OCT-1996
DEFINITIONOMEGA-3 FATTY ACID DESATURASE, ENDOPLASMIC RETICULUM.
ACCESSION P48626
PID 81345975
VERSION P48626 GI:1345975
~ DBSOURCE swissprot: locus FD3E-TOBAC, accession P48626;
class: standard.
created: Feb 1, 1996.
sequence updated: Feb l, 1996.
annotation updated: Oct 1, 1996.
3S xrefs: 8i: 1311480, 8i: 599592
KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESISE ENDOPLASMIC
RETICULUM;
TRANSMEMBRANE.
SOURCE common tobacco.
4~ ORGANISM Nicotiana tabacum
47
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Eukaryotae; Viridiplantae; Charophyta/Embryophyta group;
Embryophyta; Tracheophyta; seed plants; Magnoliophyta;
eudicotyledons; Asteridae; Solananae; Solanales; Solanaceae;
Nicotiana.
S REFERENCE 1 (residues 1 to 379)
AUTHORS Hamada,T., Kodama,H., Nishimura,M. and Iba,K.
TITLE Cloning of a cDNA encoding tobacco omega-3 fatty
acid
desaturase
JOURNAL Gene 147 (2), 293-294 (1994)
MEDLINE 95011632
REMARK SEQUENCE FROM N.A.
STRAIN=CV. SR1; TISSUE=LEAF
COMMENT [FUNCTION] ER (MICROSOMAL) OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS
OF 18:3
IS FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT MEMBRANES.
IT IS
THOUGHT TO USE CYTOCHROME B5 AS AN ELECTRON DONOR
AND TO ACT
ON FATTY ACIDS ESTERIFIED TO PHOSPHATIDYLCHOLINE
AND,
POSSIBLY, OTHER PHOSPHOLIPIDS.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
2O [SUBCELLULAR LOCATION] ENDOPLASMIC RETICULUM. [DOMAIN]
THE
HISTIDINE BOX DOMAINS MAY CONTAIN THE ACTIVE SITE
AND/OR BE INVOLVED IN METAL ION BINDING. [SIMILARITY]
TO
OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
FEATURES Location/Qualifiers
2S source 1..379
/organism="Nicotiana tabacum"
/db xref="taxon:4097"
1..379
Protei n 1..379
3O /product="OMEGA-3 FATTY ACID DESATURASE, ENDOPLASMIC
RETICULUM"
/EC number="1.14.99.-"
Region 52..72
/region name="Transmembrane region"
3S Region 97..101
/note="HISTIDINE BOX 1."
/region name="Domain"
Region 133..137
/note="HISTIDINE BOX 2."
40 /region name="Domain"
48
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Region 213..233
/region name="Transmembrane region"
Region 236..256
/region name="Transmembrane region"
Region 300..304
/note="HISTIDINE BOX 3."
/region name="Domain"
ORIGIN (SEQ ID N0; 22)
fdpsapppf rlaeirnvip khcwvkdplr slsyvvrdvi fvatligiai hldswlfypl
1~ ywaiqgtmfw aifvlghdcg hgsfsdsqll nnvvghilhs ailvpyhgwr ishkthhqnh
gnvetdeswv pmpeklynkv gystkflryk ipfpllaypm ylmkrspgks gshfnpysdl
fqpherkyvv tstlcwtvma alllylctaf gslqmfkiyg apylifvmwl dfvtylhhhg
yekklpwyrg kewsylrggl ttvdrdyglf nnihhdigth vihhlfpqip hyhlreatka
akpvlgkyyr epkksgpipf hlvkdltrsm kqdhyvsdsg eivfyqtdph if
AAD13527
(Vernicia
fordit~
LOCUS AAD13527 437 as PLN 08-FEB-1999
DEFINITION omega-3 fatty acid desaturase precursor.
ACCESSION AAD13527
~ PID g4240385
VERSION AAD13527.1 GI:4240385
DBSOURCE locus AF061027 accession AF061027.1
KEYWORDS
SOURCE Vernicia fordii.
ORGANISM Vernicia fordii
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Rosidae; eurosids
I;
Malpighiales; Euphorbiaceae; Vernicia.
~ REFERENCE 1 (residues 1 to 437)
AUTHORS Tang,F., Dyer,J.M., Lax,A.R., Shih,D.S., Chapital,D.C.
and
Pepperman,A.B.
TITLE Nucleotide sequence of a cDNA clone for omega-3
fatty acid
desaturase (Accession No. AF061027) from Aleurites
fordii
seeds (PGR99-009)
JOURNAL Plant Physiol. 119, 364 (1999)
REFERENCE 2 (residues 1 to 437)
AUTHORS Tang, F., Dyer,J.M., Lax,A.R., Shih,D.S. and Pepperman,A.B.
TITLE Direct Submission
4~ JOURNAL Submitted (21-APR-1998) Southern Regional Research
Center,
49
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USDA-ARS, 1100 Robert E. Lee Blvd., New Orleans, LA 70124,
USA
COMMENT Method: conceptual translation.
FEATURES Location/Qualifiers
source 1..437
/organism="Vernicia fordii"
/db xref="taxon:73154"
/tissue type="seeds"
Protei n <1..437
1~ /product="omega-3 fatty acid desaturase precursor"
CDS 1..437
/coded by="AF061027.1:<1..1316"
ORIGIN (SEQID N0: 23)
ereegin giegeet efdpgapppf klsdireaip khcwvkdpwr smsyvvrdva
gvi
vvfglaaaaa ylnnwivwpl ywaaqgtmfw alfvlghdcg hgsfshnpkl nsvvghllhs
silvpyhgwr ishrthhqnh ghvendeswq plsekifrsl dymtrtlrft vpspmlaypf
ylwnrspgkt gshfhpdsdl fgpnerkdvi tstvcwtama allvglslvm gpiqllklyg
mpywifvmwl dfvtylhhhg heeklpwyrg newsylrggl ttlgrdygwi nnihhdigth
vihhffpqip hyhlidatea skpvlgkyyr epdksgplsf hligylirsl kkdhyvsdtg
2~ dvvyyqtdpq 1
AAB72241
(Petroselinum
crispum)
LOCUS AAB72241 438 as PLN 08-OCT-1997
DEFINITION omega-3 fatty acid desaturase.
ACCESSION AAB72241
PID 81786066
VERSION AAB72241.1 GI:1~786066
DBSOURCE locus PCU75745 accession U75745.1
KEYWORDS
~ SOURCE parsley.
ORGANISM Petroselinum crispum
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Asteridae; euasterids
II;
Apiales; Apiaceae; Petroselinum.
REFERENCE 1 (residues 1 to 438)
AUTHORS Kirsch,C., Takamiya-Wik,M., Reinold,S., Hahlbrock,K.
and
Somssich,I.E.
TITLE Rapid, transient, and highly localized induction
of
4~ plastidial omega-3 fatty acid desaturase mRNA at
fungal
CA 02386111 2002-03-28
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infection sites in Petroselinum crispum
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 94 (5), 2079-2084
(1997)
MEDLINE 97203190
REFERENCE 2 (residues 1 to 438)
AUTHORS Somssich,I.E. and Kirsch, C.
TITLE Direct Submission
JOURNAL Submitted (23-OCT-1996) Biochemistry, Max-Planck-Institut
f.
Zuchtungsforschung, Carl-von-Linne-Weg 10, Koln,
NRW 50829,
Germany
COMMENT Method: conceptual translation supplied by author.
FEATURES Location/Qualifiers
source 1..438
/organism="Petroselinum crispum"
/db xref="taxon:4043"
/cell type="cultured parsley cells"
/clone="15-1 and 25-2"
/note="derived from two overlapping partial cDNAs"
Protein 1..438
/product="omega-3 fatty acid desaturase"
CDS 1..438
/coded by="U75745.1:96..1412"
/note="complements the Arabidopsis fad7/8 fatty
acid
double mutant"
ORIGIN (SEQ ID N0:24)
a enefdpgaap
pfklsdvraa
ipkhcwvkdp
vrsmsyvlrd
vlivfglava
asfvnnwavw
plywiaqgtmfwalfvlghd cghgsfsndaklnsvvghilhssilvpyhgwrishrthhq
nhghvendeswhplseklfn slddltrkfrftlpfpmlaypfylwgrspgkkgshydpss
dlfvpnerkdvitstvcwta maallvglnfvmgpvkmlmlygipywifvmwldfvtylhh
hghddklpwyrgkewsylrg glttldrdygwinnihhdigthvvhhlfpqiphyhlieat
eaakpvfgkyyrepkksgpv pfhllatlwksfkkdhfvsdtgdvvyyqahpe
P48625
(Glycine
max)
LOCUS FD3E SOYBN 380 as PLN O1-OCT-1996
DEFINITIONOMEGA-3 FATTY ACID SATURASE,ENDOPLASMICRETICULUM.
DE
ACCESSION P48625
3$ PID g1345974
VERSION P48625 GI:1395974
DBSOURCE swissprot: locus SOYBN,
FD3E accession
P48625;
class: standard.
created: Feb 1,
1996.
sequence updated: 1, 1996.
Feb
51
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annotation updated: Oct 1, 1996.
xrefs: gi: 408793, gi: 408794, gi: 541946
KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; ENDOPLASMIC
RETICULUM; TRANSMEMBRANE.
$ SOURCE soybean.
ORGANISM Glycine max
Eukaryotae; Viridiplantae; Charophyta/Embryophyta
group;
Embryophyta; Tracheophyta; seed plants; Magnoliophyta;
eudicotyledons; Rosidae; Fabales; Fabaceae; Papilionoideae;
1~ Glycine.
REFERENCE 1 (residues 1 to 380)
AUTHORS YADAV,N.S., WIERZBICKI,A., AEGERTER,M., CASTER,C.S.,
PEREZ-
GRAU,L., KINNEY,A.J., HITZ,W.D., BOOTH,J.R. JR.,
SCHWEIGER,B., STECCA,K.L., ALLEN,S.M., BLACKWELL,M.,
IS REITER,R.S., CARLSON,T.J., RUSSELL,S.H., FELDMANN,K.A.,
PIERCE, J. and BROWSE, J.
TITLE Cloning of higher plant omega-3 fatty acid desaturases
JOURNAL Plant Physiol. 103 (2), 467-476 (1993)
MEDLINE 94302147
2O REMARK SEQUENCE FROM N.A.
TISSUE=SEED
COMMENT [FUNCTION] MICROSOMAL (ER) OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS
OF 18:3
FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT MEMBRANES.
IT IS
2S THOUGHT TO USE CYTOCHROME B5 AS AN ELECTRON DONOR
AND TO ACT
ON FATTY ACIDS ESTERIFIED TO PHOSPHATIDYLCHOLINE
AND,
POSSIBLY, OTHER PHOSPHOLIPIDS.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
[SUBCELLULAR LOCATION] ENDOPLASMIC RETICULUM. [DOMAIN]
THE
3O HISTIDINE BOX DOMAINS MAY CONTAIN THE ACTIVE SITE
AND/OR BE INVOLVED IN METAL ION BINDING. [SIMILARITY]
TO
OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
FEATURES Location/Qualifiers
source 1..380
3S /organism="Glycine max"
/db xref="taxon:3847"
1..380
Protein 1..380
/product="OMEGA-3 FATTY ACID DESATURASE, ENDOPLASMIC
4O RETICULUM"
S2
CA 02386111 2002-03-28
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/EC number="1.14.99.-"
Region 55..75
/region name="Transmembrane region"
Region 100..104
S /note="HISTIDINE BOX 1."
/region name="Domain"
Region 136..140
/note="HISTIDINE BOX 2."
/region name="Domain"
1~ Region 212..232
/region name="Transmembrane region"
Region 236..256
/region name="Transmembrane region"
Region 303..307
1$ /note="HISTIDINE BOX 3."
/region name="Domain"
ORIGIN (SEQ ID N0:25)
fdpsap ppfkiaeira sipkhcwvkn pwrslsyvlr dvlviaalva aaihfdnwll wliycpiqgt
mfwalfvlgh dcghgsfsds pllnslvghi lhssilvpyh gwrishrthh qnhghiekde
2~ swvpltekiy knldsmtrli rftvpfplfv ypiylfsrsp gkegshfnpy snlfppserk
giaistlcwa tmfslliyls fitspllvlk lygipywifv mwldfvtylh hhghhqklpw
yrgkewsylr gglttvdrdy gwiynihhdi gthvihhlfp qiphyhlvea tqaakpvlgd
yyrepersap lpfhlikyli qsmrqdhfvs dtgdvvyyqt dslllhsqrd
25 P48618 (Brassica
nanus)
LOCUS FD3C BRAVA 404 as PLN O1-FEB-1996
DEFINITION OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST PRECURSOR.
ACCESSION P48618
PID 81345968
3~ VERSION P48618 GI:1345968
DBSOURCE swissprot: locus FD3C BRAVA, accession P48618;
class: standard.
created: Feb 1, 1996.
sequence updated: Feb l, 1996.
3$ annotation updated: Feb 1, 1996.
xrefs: 8i: 408489, 8i: 408490, 8i: 541916
KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; CHLOROPLAST;
MEMBRANE; TRANSIT PEPTIDE.
SOURCE rape.
~ ORGANISM Brassica napus
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Eukaryotae; Viridiplantae; Charophyta/Embryophyta
group;
Embryophyta; Tracheophyta; seed plants; Magnoliophyta;
eudicotyledons; Rosidae; Capparales; Brassicaceae;
Brassica.
REFERENCE 1 (residues 1 to 404)
S AUTHORS YADAV,N.S., WIERZBICKI,A., AEGERTER,M., CASTER,C.S.,
PEREZ-
GRAU,L., KINNEY,A.J., HITZ,W.D., BOOTH,J.R. JR.,
SCHWEIGER,B., STECCA,K.L., ALLEN,S.M., BLACKWELL,M.,
REITER,R.S., CARLSON,T.J., RUSSELL,S.H., FELDMANN,K.A.,
PIERCE, J. and BROWSE, J.
1~ TITLE Cloning of higher plant omega-3 fatty acid desaturases
JOURNAL Plant Physiol. 103 (2), 467-476 (1993)
MEDLINE 94302147
REMARK SEQUENCE FROM N.A.
TISSUE=SEED
IS COMMENT [FUNCTION] CHLOROPLAST OMEGA-3 FATTY ACID DESATURASE
INTRODUCES
THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS OF 16:3
AND 18:3
FATTY ACIDS, IMPORTANT CONSTITUENTS OF PLANT MEMBRANES.
IT IS
THOUGHT TO USE FERREDOXIN AS AN ELECTRON DONOR AND
TO ACT ON
ZO FATTY ACIDS ESTERIFIED TO GALACTOLIPIDS, SULFOLIPIDS
AND
PHOSPHATIDYLGLYCEROL.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
[SUBCELLULAR LOCATION] CHLOROPLAST, MEMBRANE-BOUND
(PROBABLE). [DOMAIN] THE HISTIDINE BOX DOMAINS MAY
CONTAIN
ZS THE ACTIVE SITE AND/OR BE INVOLVED IN METAL ION
BINDING.
[SIMILARITY] TO OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
FEATURES Location/Qualifiers
source 1..404
/organism="Brassica napus"
/db xref="taxon:3708"
1..404
Protein <1..404
/product="OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST
PRECURSOR"
3S /EC number="1.14.99.-"
Region <1..(2.404)
/note="CHLOROPLAST."
/region name="Transit peptide"
Region (1.403)..404
4O /note="OMEGA-3 FATTY ACID DESATURASE,CHLOROPLAST."
S4
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/region name="Mature chain"
Region 121..125
/note="HISTIDINE BOX 1."
/region name="Domain"
Region 157..161
/note="HISTIDINE BOX 2."
/region name="Domain"
Region 324..328
/note="HISTIDINE BOX 3."
/region name="Domain"
ORIGIN (SEQ ID NO: 26)
ieee pktqrfdpga pppfnladir aaipkhcwvk npwksmsyvv relaivfala agaaylnnwl
vwplywiaqg tmfwalfvlg hdcghgsfsn dprlnsvvgh llhssilvpy hgwrishrth
hqnhghvend eswhpmseki yksldkptrf frftlplvml aypfylwars pgkkgshyhp
dsdlflpker ndvltstacw tamavllvcl nfvmgpmqml klyvipywin vmwldfvtyl
hhhghedklp wyrgkewsyl rgglttldrd yglinnihhd igthvihhlf pqiphyhlve
ateaakpvlg kyyrepdksg plplhllgil aksikedhfv sdegdvvyye adpnly
BAA22440 (Zea mars)
2~ LOCUS BAA22440 398 as PLN 04-MAR-1998
DEFINITION fatty acid desaturase.
ACCESSION BAA22440
PID g2446996
VERSION BAA22440.1 GI:2446996
DBSOURCE locus D63953 accession D63953.1
KEYWORDS
SOURCE Zea mays.
ORGANISM Zea mays
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
Liliopsida; Poales; Poaceae; Zea.
REFERENCE 1 (residues 1 to 398)
AUTHORS Kusano,T.
TITLE Direct Submission
3$ JOURNAL Submitted (30-AUG-1995) to the DDBJ/EMBL/GenBank databases.
Tomonobu Kusano, Akita Prefectural College of Agriculture,
Biotechnology Institute; 2-2 Minami, Ohgatamura, Minamiakita-
gun, Akita 010-04, Japan (E-mail:kusano@air.akita-u. ac.jp,
Te1:0185-45-2026(ex.403), Fax:0185-45-2678)
~ REFERENCE 2 (sites)
CA 02386111 2002-03-28
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AUTHORS Berberich,T., Harada,M., Sugawara,K., Kodama,H.,
Iba,K. and
Kusano,T.
TITLE Two maize genes encoding omega-3 fatty acid desaturase
and
their differential expression to temperature
$ JOURNAL Plant Mol. Biol. 36 (2), 297-306 (1998)
MEDLINE 98145435
COMMENT Sequence updated (11-Apr-1996) by: Tomonobu Kusano.
FEATURES Location/Qualifiers
source 1..398
1~ /organism="Zea mays"
/strain="honey bantum"
/db xref="taxon:4577"
Protei n 1..398
/product="fatty acid desaturase"
1$ CDS 1..398
/gene="FAD8"
/coded by="D63953.1:<1..1198"
ORIGIN (SEQID N0: 27) '
veedkr ssplgegdeh vaasgaagge fdpgapppfg laeiraaipk hcwvkdpwrs
mayvlrdvvv
2~ vlglaaaaar ldswlvwply waaqgtmfwa lfvlghdcgh gsfsnnpkln svvghilhss
ilvpyhgwri shrthhqnhg hvekdeswhp lperlyksld fmtrklrftm pfpllafply
lfarspgksg shfnpssdlf qpnekkdiit staswlamvg vlagltflmg pvamlklygv
pyfvfvawld mvtylhhhgh edklpwyrgq ewsylrgglt tldrdyglin nihhdigthv
ihhlfpqiph yhlieateaa kpvlgkyyke pkksgplpwh lfgvlaqslk qdhyvsdtgd
vvyyqtd
2$
P48621 (Glycine
max)
LOCUS FD3C SOYBN 453 as PLN 15-DEC-1998
DEFINITION OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST PRECURSOR.
ACCESSION P48621
30 PID 81345971
VERSION P48621 GI:1345971
DBSOURCE swissprot: locus FD3C SOYBN, accession P48621;
class: standard.
created: Feb 1, 1996.
3$ sequence updated: Feb 1, 1996.
annotation updated: Dec 15, 1998.
xrefs: 8i: 408791, 8i: 408792, 8i: 541947
xrefs (non-sequence databases): PFAM PF00487
KEYWORDS OXIDOREDUCTASE; FATTY ACID BIOSYNTHESIS; CHLOROPLAST;
4O MEMBRANE; TRANSIT PEPTIDE.
$6
CA 02386111 2002-03-28
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SOURCE soybean.
ORGANISM Glycine max
Eukaryota; Viridiplantae; Charophyta/Embryophyta
group;
Embryophyta; Tracheophyta; euphyllophytes; Spermatophyta;
Magnoliophyta; eudicotyledons; Rosidae; Fabales;
Fabaceae;
Papilionoideae; Glycine.
REFERENCE 1 (residues 1 to 453)
AUTHORS YADAV,N.S., WIERZBICKI,A., AEGERTER,M., CASTER,C.S.,
PEREZ-
GRAU,L., KINNEY,A.J., HITZ,W.D., BOOTH,J.R. JR.,
IO SCHWEIGER,B., STECCA,K.L., ALLEN,S.M., BLACKWELL,M.,
REITER,R.S., CARLSON,T.J., RUSSELL,S.H., FELDNIANN,K.A.,
PIERCE,J. and BROWSE J.
TITLE Cloning of higher plant omega-3 fatty acid desaturases
JOURNAL Plant Physiol. 103 (2), 467-476 (1993)
IS MEDLINE 94302147
REMARK SEQUENCE FROM N.A.
TISSUE=SEED
COMMENT [FUNCTION] CHLOROPLAST OMEGA-3 FATTY ACID DESATURASE
INTRODUCES THE THIRD DOUBLEBOND IN THE BIOSYNTHESIS
OF 16:3
ZO AND 18:3 FATTY ACIDS, IMPORTANT CONSTITUENTS OF
PLANT
MEMBRANES. IT IS THOUGHT TO USE FERREDOXIN AS AN
ELECTRON
DONOR AND TO ACT ON FATTY ACIDS ESTERIFIED TO GALACTOLIPIDS,
SULFOLIPIDS AND PHOSPHATIDYLGLYCEROL.
[PATHWAY] POLYUNSATURATED FATTY ACID BIOSYNTHESIS.
ZS [SUBCELLULAR LOCATION] CHLOROPLAST, MEMBRANE-BOUND
(PROBABLE). [DOMAIN] THE HISTIDINE BOX DOMAINS MAY
CONTAIN
THE ACTIVE SITE AND/OR BE INVOLVED IN METAL ION
BINDING.
[SIMILARITY] TO OTHER PLANT OMEGA-3 FATTY ACID DESATURASES.
FEATURES Location/Qualifiers
30 source 1..453
/organism="Glycine max"
/db xref="taxon:3897"
1..453
Protein 1..453
3S /product="OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST
PRECURSOR"
/EC number="1.14.99.-"
Region 1..(2.453)
/region name="Transit peptide"
4O /note="CHLOROPLAST."
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Region (1.452)..453
/region name="Mature chain"
/note="OMEGA-3 FATTY ACID DESATURASE, CHLOROPLAST."
Region 171..175
$ /region name="Domain"
/note="HISTIDINE BOX 1."
Region 207..211
/region name="Domain"
/note="HISTIDINE BOX 2."
Region 374..378
/region name="Domain"
/note="HISTIDINE BOX 3."
ORIGIN (SEQ ID NO: 28)
svd ltngtngveh eklpefdpga pppfnladir aaipkhcwvk dpwrsmsyvv rdviavfgla
aaaaylnnwl vwplywaaqg tmfwalfvlg hdcghgsfsn nsklnsvvgh llhssilvpy
hgwrishrth hqhhghaend eswhplpekl frsldtvtrm lrftapfpll afpvylfsrs
pgktgshfdp ssdlfvpner kdvitstacw aamlgllvgl gfvmgpiqll klygvpyvif
vmwldlvtyl hhhghedklp wyrgkewsyl rgglttldrd ygwinnihhd igthvihhlf
pqiphyhlve ateaakpvfg kyyrepkksa aplpfhlige iirsfktdhf vsdtgdvvyy qtd
BAA22441 (Zea mans)
LOCUS BAA22441 443 as PLN 04-MAR-1998
DEFINITION fatty acid desaturase.
ACCESSION BAA22441
PID 82446998
VERSION BAA22441.1 GI:2446998
DBSOURCE locus D63954 accession D63954.1
KEYWORDS
SOURCE Zea mays.
ORGANISM Zea mays Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
Liliopsida; Poales; Poaceae; Zea.
REFERENCE 1 (residues 1 to 443)
AUTHORS Kusano,T.
3$ TITLE Direct Submission
JOURNAL Submitted (30-AUG-1995) to the DDBJ/EMBL/GenBank
databases. Tomonobu Kusano, Akita Prefectural College of
Agriculture, Biotechnology Institute; 2-2 Minami, Ohgatamura,
Minamiakita-
gun, Akita 010-04, Japan (E-mail:kusano@air.akita-u. ac.jp,
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Te1:0185-45-2026(ex.403), Fax:0185-45-2678)
REFERENCE 2 (sites)
AUTHORS Berberich,T., Harada,M., Sugawara,K., Kodama,H., Iba,K. and
Kusano,T.
$ TITLE Two maize genes encoding omega-3 fatty acid desaturase and
their differential expression to temperature
JOURNAL Plant Mol. Biol. 36 (2), 297-306 (1998)
MEDLINE 98145435
FEATURES Location/Qualifiers
1~ source 1..443
/organism="Zea mays"
/strain="honey bantum"
/db xref="taxon:4577"
Protein 1..443
15 /product="fatty acid desaturase"
CDS 1..443
/gene="FAD7"
/coded by="join(D63954.1:2178..2665,D63954.1:2775..2
20 864,
D63954.1:2944..3010,D63954.1:3113..3205,
D63954.1:3323..3508,D63954.1:3615..3695,
D63954.1:4259..4396,D63959.1:4492..4680)"
ORIGIN (SEQ ID N0: 29)
25 ga aaggefdpga pppfglaeir aaipkhcwvk dpwrsmsyvl rdvavvlgla aaaarldswl
vwplywaaqg tmfwalfvlg hdcghgsfsn npklnsvvgh ilhssilvpy hgwrishrth
hqnhghvekd eswhplperl yksldfmtrk lrftmpfpll afplylfars pgksgshfnp
gsdlfqptek ndiitstasw lamvgvlagl tflmgpvpml klygvpylvf vawldmvtyl
hhhghedklp wyrgkewsyl rgglttldrd ygwinnihhd igthvihhlf pqiphyhlie
30 ateaakpvlg kyykepknsg alpwhlfrvl aqslkqdhyv shtgdvvyyq ae
CAA07638 (Solarium tuberosum)
LOCUS CAA07638 431 as PLN 04-SEP-1998
DEFINITION w-3 desaturase.
3S ACCESSION CAA07638
PID 83550663
VERSION CAA07638.1 GI:3550663
DBSOURCE embl locus STU007739, accession AJ007739.1
KEYWORDS
~ SOURCE potato.
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ORGANISM Solanum tuberosum
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; Asteridae; Solananae; Solanales; Solanaceae;
Solanum; Potatoes section Petota.
REFERENCE 1 (residues 1 to 431)
AUTHORS Leon, J.
TITLE Direct Submission
JOURNAL Submitted (20-AUG-1998) Leon J., Genetica
Molecular de PLantas, Centro Nacional de Biotecnologia
(CSIC), Campus de Cantoblanco Ctra. Colmenar Viejo Km 15,500,
Madrid 28049, SPAIN
REFERENCE 2 (residues 1 to 431)
AUTHORS Martin, M.
JOURNAL Unpublished
FEATURES Location/Qualifiers
source 1..431
/organism="Solanum tuberosum"
/cultivar="Desiree"
/db xref="taxon:4113"
Protein 1..431
/product="w-3 desaturase"
CDS 1..431
/db xref="SPTREMBL:082068"
/coded by="AJ007739.1:1..1296"
ORIGIN (SEQ ID N0: 30)
eeeqt tnngdefdpg asppfklsdi kaaipkhcwv knpwtsmsyv vrdvaivfgl aaaaayfnnw
lvwplywfaq stmfwalfvl ghdcghgsfs nnhnlnsvag hilhssilvp
yhgwrishrt hhqnhghven deswhplsek lynsldditk kfrftlpfpl laypfylwgr
spgkkgshfd pssdlfvase kkdvitstvc wtamaallvg lsfvmgplqv lklygipywg
fvmwldivty lhhhghedkv pwyrgeewsy lrgglttldr dygwinnihh digthvihhl
fpqiphyhlv eateaakpvl gkyykepkks gplpfyllgy liksmkedhf vsdtgnvvyy
qtdpnly
AAA86690 (Limnanthes dou~lasial
LOCUS AAA86690 436 as PLN 21-NOV-1995
DEFINITION delta-15 lineoyl desaturase.
ACCESSION AAA86690
PID 8699390
VERSION AAA86690.1 GI:699390
CA 02386111 2002-03-28
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DBSOURCE locus LDU17063 accession U17063.1
KEYWORDS
SOURCE Douglas's meadowfoam.
ORGANISM Limnanthes douglasii
$ Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Rosidae; eurosids
II;
Brassicales; Limnanthaceae; Limnanthes.
REFERENCE 1 (residues 1 to 436)
1~ AUTHORS Bhella,R.S. and MacKenzie,S.L.
TITLE Nucleotide sequence of a cDNA from Limnanthes douglasii
L.
Encoding a delta-15 linoleic acid desaturase
JOURNAL Plant Physiol. 108 (2), 861 (1995)
MEDLINE 95334518
1S REFERENCE 2 (residues 1 to 436)
AUTHORS MacKenzie,S.L.
TITLE Direct Submission
JOURNAL Submitted (09-NOV-1994) Samuel L. MacKenzie, Plant
Biotechnology Institute, National Research Council
of Canada,
110 Gymnasium Place, Saskatoon, SK S7N OW9, Canada
COMMENT Method: conceptual translation.
FEATURES Location/Qualifiers
source 1..436
/organism="Limnanthes douglasii"
25 /db xref="taxon:28973"
/dev stage="seed, storage deposition stage"
Protei n 1..436
/product="delta-15 lineoyl desaturase"
CDS 1..436
/function="linoleic acid desaturation"
/coded by="U17063.1:56..1366"
/note="omega-3-fatty acid desaturase"
ORIGIN
(SEQ ID
N0: 31)
v sapfqiastt peeedevaef dpgspppfkl adiraaipkh cwvknqwrsm syvvrdvviv
3$ lglaaaavaanswavwplyw vaqgtmfwal fvlghdcghg sfsnnhklns vvghllhssi
lvpyhgwrirhrthhqnhgh vendeswhpm seklfrsldk ialtfrfkap fpmlaypfyl
werspgktgshyhpdsdlfv psekkdvits ticwttmvgl liglsfvmgp iqilklyvvp
ywifvmwldfvtyldhhghe dklpwyrgee wsylrggltt ldrdyglinn ihhdigthvi
hhlfpqiphyhlveatqaak pifgkyykep akskplpfhl idvllkslkr dhfvpdtgdi
4~ vyyqsdpq
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BAA07785 (Triticum aestivum)
LOCUS BAA07785 380 as PLN 18-JUN-1999
DEFINITION plastid omega-3 fatty acid desaturase.
S ACCESSION BAA07785
PID 81694615
VERSION BAA07785.1 GI:1694615
DBSOURCE locus D43688 accession D43688.1
KEYWORDS
1~ SOURCE bread wheat.
ORGANISM Triticum aestivum
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
Liliopsida; Poales; Poaceae; Triticum
REFERENCE 1 (sites)
AUTHORS Horiguchi,G., Iwakawa,H., Kodama,H., Kawakami,N.,
Nishimura,M.
And Iba,K.
2~ TITLE Expression of a gene for plastid omega-3 fatty acid
desaturase and changes in lipid and fatty acid compositions
in light- and dark-grown wheat leaves
JOURNAL Physiol. Plantarum 96, 275-283 (1996)
REFERENCE 2 (residues 1 to 380)
AUTHORS Iwakawa,H.
TITLE Direct Submission
JOURNAL Submitted (03-DEC-1994) to the DDBJ/EMBL/GenBank
databases. Hirotaka Iwakawa, Kyushu University,
Facul.
Science, Dept. Biology, Lab. Plant Physiology; 6-10-1
Hakozaki, Higashi-ku,
Fukuoka, Fukuoka 812, Japan (E-mail: koibascb@mbox.nc.kyushu-
u.ac.jp, Te1:092-641-1101(ex.4414), Fax:092-632-2741)
FEATURES Location/Qualifiers
source 1..380
/organism="Triticum aestivum"
/strain="cv. Chihoku"
/db xref="taxon:4565"
/clone lib="lambda-gtll"
/tissue type="leaf"
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Protein 1..380
/product="plastid omega-3 fatty acid desaturase"
CDS 1..380
/gene="TaFAD7"
$ /coded by="D43688.1:<1..1143"
ORIGIN (SEQ ID N0: 32)
fdpgapp pfgladiraa ipkhcwvkdh wssmgyvvrd vvvvlalaat aarldswlaw pvywaaqgtm
fwalfvlghd cghgsfsnna klnsvvghil hssilvpyng wrishrthhq nhghvendes
whplpeklyr sldsstrklr falpfpmlay pfylwsrspg ksgshfhpss dlfqpnekkd
1~ iltsttcwla magllagltv vmgplqilkl yavpywifvm wldfvtylhh hghndklpwy
rgkawsiytg glttldrdyg wlnnihhdig thvihhllpq iphyhlveat eaatvlgkyy
repdksgpfp fhlfgalars mksdhyvsdt gdiiyyqtdp k
BAA28358
(Triticum
aestivum)
15 LOCUS BAA28358 383 as PLN 30-MAY-1998
DEFINITION omega-3 fatty acid desaturase.
ACCESSION BAA28358
PID 83157460
VERSION BAA28358.1 GI:3157460
~ DBSOURCE locus D84678 accession D84678.1
KEYWORDS
SOURCE Triticum aestivum.
ORGANISM Triticum aestivum
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
2$ Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
Liliopsida; Poales; Poaceae; Triticum.
REFERENCE 1 (residues 1 to 383)
AUTHORS Horiguchi,G.
TITLE Direct Submission
3~ JOURNAL Submitted (O1-MAY-1996) to the DDBJ/EMBL/GenBank
databases.
Gorou Horiguchi, Kyushu University, Faculty of Science,
Department of Biology; 6-10-1 Hakozaki, Fukuoka,
Fukuoka 812-
8581, Japan (E-mail:ghoriscb@mbox.nc.kyushu-u. ac.jp,
Te1:092-
642-2621, Fax:092-642-2621)
3$ REFERENCE 2 (sites)
AUTHORS Horiguchi,G., Kawakami,N., Kusumi,K., Kodama,H.
and Iba,K.
TITLE Developmental regulation of genes for microsome
and plastid
omega-3 fatty acid desaturases in wheat (Triticum
aestivum
L.)
4~ JOURNAL Plant Cell Physiol. 39, 540-544 (1998)
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FEATURES Location/Qualifiers
source 1..383
/organism="Triticum aestivum"
/cultivar="Chihoku"
$ /db xref="taxon:4565"
/clone="pWFD3"
/clone lib="lambda MOSE lox"
/tissue type="leaf and root"
Protein
1..383
1~ /product="omega-3 fatty acid desaturase"
CDS 1..383
/gene="TaFAD3"
/coded by="D84678.1:132..1283"
ORIGIN
(SEQ ID
N0: 33)
1$ fdaakppp
frigdvraav
pahcwpqepp
aslsyvardv
avvaalaaaa
wradswalwp
lywavqgtmfwalfvlghdc ghgsfsdsgt lnsvvghllh tfilvpyngw rishrthhqn
hghidrdeswhpitekvyqk leprtktlrf svpfpllafp vylwyrspgk egshfnpssd
lftpkerrdviisttcwftm ialligmacv fglvpvlkly gvpyivnvmw ldlvtylhhh
ghqdlpwyrgeewsylrggl ttvdrdygwi nnihhdigth vihhlfpqip hyhlveatka
~ arpvlgryyrepeksgplpm hlitvllksl rvdhfvsdvg dvvfyqtdps 1
BAA11397
(Oryza
sativa)
LOCUS BAA11397 381 as PLN 05-FEB-1999
DEFINITIONw-3 fatty acid desaturase.
2$ ACCESSION BAA11397
PID 81777376
VERSION BAA11397.1 GI:1777376
DBSOURCE locus RICP181X2 accession D78506.1
KEYWORDS
30 SOURCE Oryza sativa.
ORGANISM Oryza sativa
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
Liliopsida;
3$ Poales; Poaceae; Oryza.
REFERENCE 1 (residues 1 to 381)
AUTHORS Akagi,H.
TITLE Direct Submission
JOURNAL Submitted (27-NOV-1995) to the DDBJ/EMBL/GenBank
4~ databases. Hiromori Akagi, Life Science Institute,
Mitsui
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Toatsu Chemicals Inc., Plant Biothechnology; Togo
1144,
Mobara, Chiba 297, Japan
(E-mail:tnirasaw@niguts.nig. ac.jp, Te1:0475-25-6729,
Fax:0475-25-6553)
$ REFERENCE 2 (residues 1 to 381)
AUTHORS Akagi,H.
TITLE Nucleotide sequence of a w-3 fatty acid desaturase
gene of
rice
JOURNAL Unpublished (1996)
IO REFERENCE 3 (sites)
AUTHORS Kodama,H., Akagi,H., Kusumi,K., Fujimura,T. and Iba,K.
TITLE Structure, chromosomal location and expression of
a rice gene
encoding the microsome omega-3 fatty acid desaturase
JOURNAL Plant Mol. Biol. 33 (3), 493-502 (1997)
15 MEDLINE 97201483
FEATURES Location/Qualifiers
source 1..381
/organism="Oryza sativa"
/strain="IR36"
/db xref="taxon:4530"
/clone="p18-1X2"
Protei n 1..381
/product="w-3 fatty acid desaturase"
CDS 1..381
25 /coded by="join(D78506.1:674..975,D78506.1:1069..115
8, D78506.1:1613..1679,D78506.1:2499..2582,
D78506.1:2741..2926,D78506.1:3030..3107,
D78506.1:3662..3799,D78506.1:3917..4117)"
ORIGIN ID N0:34)
(SEQ
~ sedarlf akpppfr igdvraaipv hcwrktplrs lsyvardlli vaalfaaaas
fda sidlawawaw
plywarqgtmvwalfvlghd cghgsfsdsa mlnnvvghll hsfilvpyhg wrfshrthhq
nhghierdeswhpiteklyw qletrtkklr ftlpftllaf pwyrspgktg shflpssdlf
spkeksdvivsttcwcimis llvalacvfg pvpvlmlygv pylvfvmwld lvtylhhhgh
ndlpwyrgeewsylrggltt vdrdygwinn ihhdigthvi hhlfpqiphy hlveatkaar
3$ pvlgryyrepeksgplplhl fgvllrtlrv dhfvsdvgdv vyyqtdhsl
AAB61352 (Synechococcus PCC7002)
LOCUS AAB61352 350 as BCT 17-JUN-1997
DEFINITION omega-3 desaturase.
~ ACCESSION AAB61352
CA 02386111 2002-03-28
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PID g2197199
VERSION AAB61352.1 GI:2197199
DBSOURCE locus SPU36389 accession U36389.1
KEYWORDS
SOURCE Synechococcus PCC7002.
ORGANISM Synechococcus PCC7002
Bacteria; Cyanobacteria; Chroococcales; Synechococcus.
REFERENCE 1 (residues 1 to 350)
AUTHORS Sakamoto,T, and Bryant,D.A.
TITLE Temperature-regulated mRNA accumulation and stabilization
for
Fatty acid desaturase genes in the cyanobacterium
Synechococcus sp.strain PCC 7002
JOURNAL Mol. Microbiol. 23 (6), 1281-1292 (1997)
MEDLINE 97260123
IS REFERENCE 2 (residues 1 to 350)
AUTHORS Sakamoto,T.
TITLE Direct Submission
JOURNAL Submitted (14-SEP-1995) Toshio Sakamoto, Biochemistry
and
Molecular Biology, The Pennsylvania State University,
S-232
Frear Bldg., University Park, PA 16802, USA
FEATURES Location/Qualifiers
source 1..350
/organism="Synechococcus PCC7002"
/db xref="taxon:32049"
Protei n 1..350
/function="desaturation of fatty acids at omega-3
position"
/product="omega-3 desaturase"
CDS 1..350
/gene="desB"
/coded by="U36389.1:747..1799"
/transl table=11
ORIGIN
(SEQ ID
N0: 35)
pf tlkdvkaaip dycfqpsvfr slayffldig iiaglyaiaa yldswffypi fwfaqgtmfw
alfvvghdcghgsfsrskfl ndlighlsht pilvpfhgwr ishrthhsnt gnidtdeswy
pipeskydqmgfaeklvrfy apliaypiyl fkrspgrgpg shfspksplf kpaerndiil
staaiiamvgflgwftvqfg llafvkfyfv pyvifviwld lvtylhhtea dipwyrgddw
yylkgalstidrdygifnei hhnigthvah hifhtiphyh lkdateaikp llgdyyrvsh
apiwrsffrsqkachyiadq gshlyyq
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552650
(Synechocystis
sp.)
LOCUS 552650 359 as BCT 13-MAR-1997
DEFINITIONdesaturase delta 15 - Synechocystis sp. (strain
PCC6803).
ACCESSION 552650
$ PID 82126522
VERSION S52650 GI:2126522
DBSOURCE pir: locus 552650;
summary: #length 359 #molecular-weight 41919 #checksum
9162;
genetic: #start Codon GTG;
1~ PIR dates: 28-Oct-1996 #sequence_revision 13-Mar-1997
#text change 13-Mar-1997.
KEYWORDS
SOURCE Synechocystis sp.
ORGANISM Synechocystis sp.
1$ Eubacteria; Cyanobacteria; Chroococcales; Synechocystis.
REFERENCE 1 (residues 1 to 359)
AUTHORS Sakamoto,T., Los,D.A., Higashi,S., Wada,H., Nishida,I.,
Ohmori,M. and Murata,N.
TITLE Cloning of omega 3 desaturase from cyanobacteria
and its use
in altering the degree of membrane-lipid unsaturation
JOURNAL Plant Mol. Biol. 26 (1), 249-263 (1994)
MEDLINE 95035996
FEATURES Location/Qualifiers
source 1..359
25 /organism="Synechocystis sp."
/db xref="taxon:1143"
Protei n 1..359
/product="desaturase delta 15"
ORIGIN
(SEQ ID
N0: 36)
~ pftlqelrnaipadcfepsv vrslgyffld vgliagfyal aayldswffy pifwliqgtl
fwslfvvghdcghgsfsksk tlnnwighls htpilvpyhg wrishrthha ntgnidtdes
wypvseqkynqmawyekllr fylpliaypi ylfrrspnrq gshfmpgspl frpgekaavl
tstfalaafvgflgfltwqf gwlfllkfyv apylvfvvwl dlvtflhhte dnipwyrgdd
wyflkgalstidrdygfinp ihhdigthva hhifsnmphy klrrateaik pilgeyyrys
35 depiwqaffksywachfvpn qgsgvyyqs
AAA61774
(Chloroplast
Brassica
napus)
LOCUS AAA61774 329 as PLN 31-JAN-1995
DEFINITIONomega-3 fatty acid desaturase.
ACCESSION AAA61774
4~ PID 8408490
67
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VERSION AAA61774.1 GI:408490
DBSOURCE locus BNACPFADD accession L22963.1
KEYWORDS
SOURCE rape.
$ ORGANISM Chloroplast Brassica napus
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Rosidae; eurosids
II;
Brassicales; Brassicaceae; Brassica.
1~ REFERENCE 1 (residues 1 to 329)
AUTHORS Yadav,N.S., Wierzbicki,A., Aegerter,M., Caster,C.S.,
Perez-
Grau,L., Kinney,A.J., Hitz,W.D., Booth,J.R. Jr.,
Schweiger,B., Stecca,K.L.
TITLE Cloning of higher plant omega-3 fatty acid desaturases
1$ JOURNAL Plant Physiol. 103 (2), 467-476 (1993)
MEDLINE 94302147
COMMENT Method: conceptual translation.
FEATURES Location/Qualifiers
source 1..329
/organism="Brassica napus"
/chloroplast
/db xref="taxon:3708"
/tissue type="seed"
Protei n 1..329
2$ /product="omega-3 fatty acid desaturase"
CDS 1..329
/gene="Fadd"
/coded by="L22963.1:226..1215"
ORIGIN ID NO: 37)
(SEQ
~ msyvvrelaivfalaagaay lnnwlvwply wiaqgtmfwa lfvlghdcgh gsfsndprln
svvghllhssilvpyhgwri shrthhqnhg hvendeswhp msekiyksld kptrffrftl
plvmlaypfylwarspgkkg shyhpdsdlf lpkerndvlt stacwtamav llvclnfvmg
pmqmlklyvipywinvmwld fvtylhhhgh edklpwyrgk ewsylrgglt tldrdyglin
nihhdigthvihhlfpqiph yhlveateaa kpvlgkyyre pdksgplplh llgilaksik
3$ edhfvsdegdvvyyeadpnl y
BAA22439 (Zea mars)
LOCUS BAA22439 262 as PLN 04-MAR-1998
DEFINITION fatty acid desaturase.
~ ACCESSION BAA22439
68
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PID 82946994
VERSION BAA22439.1 GI:2446994
DBSOURCE locus D63952 accession D63952.1
KEYWORDS
S SOURCE Zea mays.
ORGANISM Zea mays
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
Liliopsida; Poales; Poaceae; Zea.
1~ REFERENCE 1 (residues 1 to 262)
AUTHORS Kusano,T.
TITLE Direct Submission
JOURNAL Submitted (30-AUG-1995) to the DDBJ/EMBL/GenBank
databases.
Tomonobu Kusano, Akita Prefectural College of Agriculture,
IS Biotechnology Institute; 2-2 Minami, Ohgatamura,
Minamiakita-
gun, Akita 010-04, Japan (E-mail:kusano@air.akita-u.
ac.jp,
Te1:0185-45-2026(ex.403), Fax:0185-45-2678)
REFERENCE 2 (sites)
AUTHORS Berberich,T., Harada,M., Sugawara,K., Kodama,H.,
Iba,K. and
2~ Kusano,T.
TITLE Two maize genes encoding omega-3 fatty acid desaturase and
their differential expression to temperature
JOURNAL Plant Mol. Biol. 36 (2), 297-306 (1998)
MEDLINE 98145435
ZS FEATURES Location/Qualifiers
source 1..262
/organism="Zea mays"
/strain="honey bantum"
/db xref="taxon:4577"
Protein 1..262
/product="fatty
acid desaturase"
CDS 1..262
/gene="FAD7"
/coded by="D63952.1:<1..791"
3S ORIGIN (SEQ ID NO: )
38
lhssilvpyh gwrishrthhqnhghvekde swhplperlyksldfmtrkl rftmpfplla
fplylfarsp gksgshfnpgsdlfqptekn diitstaswlamvgvlaglt flmgpvpmlk
lygvpylvfv awldmvtylhhhghedklpw yrgkewsylrgglttldrdy gwinnihhdi
gthvihhlfp qiphyhlieateaakpvlgk yykepknsgalpwhlfrvla qslkqdhyvs
4~ htgdvvyyqa a
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BAA11396 (Oryza sativa)
LOCUS BAA11396 269 as PLN 05-FEB-1999
DEFINITION w-3 fatty acid desaturase.
$ ACCESSION BAA11396
PID 81785856
VERSION BAA11396.1 GI:1785856
DBSOURCE locus RICPAll accession D78505.1
KEYWORDS
1~ SOURCE Oryza sativa.
ORGANISM Oryza sativa
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
Liliopsida; Poales; Poaceae; Oryza.
IS REFERENCE 1 (residues 1 to 269)
AUTHORS Akagi,H.
TITLE ' Direct Submission
JOURNAL Submitted (27-NOV-1995) to the DDBJ/EMBL/GenBank databases.
Hiromori Akagi, Life Science Institute, Mitsui Toatsu
~ Chemicals
Inc., Plant Biothechnology; Togo 1144, Mobara, Chiba 297,
Japan
(E-mail:tnirasaw@niguts.nig. ac.jp, Te1:0475-25-6729,
Fax:0475-25-6553)
25 REFERENCE 2 (residues 1 to 269)
AUTHORS Akagi,H.
TITLE Partial nucleotide sequence of a w-3 fatty acid
desaturase
cDNA Of rice
JOURNAL Unpublished (1996)
~ REFERENCE 3 (sites)
AUTHORS Kodama,H., Akagi,H., Kusumi,K., Fujimura,T. and
Iba,K.
TITLE Structure, chromosomal location and expression of
a rice gene
encoding the microsome omega-3 fatty acid desaturase
JOURNAL Plant Mol. Biol. 33 (3), 493-502 (1997)
3S MEDLINE 97201483
COMMENT Sequence updated (20-Jan-1997) by: Hiromori Akagi.
FEATURES Location/Qualifiers
source 1..269
/organism="Oryza sativa"
/strain="Nipponbare"
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/db xref="taxon:4530"
Protein 1..269
/product="w-3 fatty acid desaturase"
CDS 1..269
$ /coded by="D78505.1:<1..810"
ORIGIN (SEQ ID N0: 39)
nnvvghllhs filvpyhgwr fshrthhqnh ghierdeswh piteklywql etrtkklrft
lpftllafpw yrspgktgsh flpssdlfsp keksdvivst tcwcimisll valacvfgpv
pvlmlygvpy lvfvmwldlv tylhhhghnd lpwyrgeews ylrgglttvd rdygwinnih
1~ hdigthvihh lfpqiphyhl veatkaarpv lgryyrepek sgplplhlfg vllrtlrvdh
fvsdvgdvvy yqtdhsl
AAD41582
(Brassica
raga)
LOCUS AF056572 1 172 as PLN O1-JUL-1999
1$ DEFINITIONunknown.
ACCESSION AAD41582
PID 85305314
VERSION AAD41582.1 GI:5305314
DBSOURCE locus AF056572 accession AF056572.1
ZO KEYWORDS
SOURCE Brassica rapa.
ORGANISM Brassica rapa
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
25 eudicotyledons; core eudicots; Rosidae; eurosids
II;
Brassicales; Brassicaceae; Brassica.
REFERENCE 1 (residues 1 to 172)
AUTHORS Brunel,D., Froger,N. and Pelletier,G.
TITLE Development of amplified consensus genetic markers
(A.C.G.M.)
in Brassica napus from Arabidopsis thaliana sequences
of
known biological function
JOURNAL Unpublished
REFERENCE 2 (residues 1 to 172)
AUTHORS Brunel,D., Froger,N. and Pelletier,G.
3$ TITLE Direct Submission
JOURNAL Submitted (O1-APR-1998) Station de Genetique et
d'Amelioration des Plantes, INRA, Route de St Cyr,
Versailles
78026, France
COMMENT Method: conceptual translation.
~ FEATURES Location/Qualifiers
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source 1..172
/organism="Brassica raga"
/cultivar="R500"
/db xref="taxon:3711"
Protein <1..>172
/product="unknown"
CDS 1..172
/gene="FAD31"
/note="similar to Arabidopsis thaliana FAD3"
1~
/coded by="join(AF056572.1:<1..26,AF056572.1:557..62
3, AF056572.1:1221..1406,
AF056572.1:1484..1564,AF056572.1:1652..>1714)"
ORIGIN (SEQ ID N0: 40)
15 filvpyhgwr ishrthhqnh ghvendeswv plpeklyknl shstrmlryt vplpmlaypl
ylwyrspgke gshynpyssl fapserklia tsttcwsiml atlvylsflv gpvtvlkvyg
vpyiifvmwl davtylhhhg hddklpwyrg kewsylrggl ttidrdygif nn
AAD41581
(Brassica
oleracea)
20 LOCUS AF056571 1 141 as PLN O1-JUL-1999
DEFINITIONunknown.
ACCESSION AAD41581
PID 85305312
VERSION AAD41581.1 GI:5305312
25 DBSOURCE locus AF056571 accession AF056571.1
KEYWORDS
SOURCE Brassica oleracea.
ORGANISM Brassica oleracea
Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Rosidae; eurosids
II;
Brassicales; Brassicaceae; Brassica.
REFERENCE 1 (residues 1 to 141)
AUTHORS Brunel,D., Froger,N. and Pelletier,G.
35 TITLE Development of amplified consensus genetic markers
(A.C.G.M.)
in Brassica napus from Arabidopsis thaliana sequences
of
known biological function
JOURNAL Unpublished
REFERENCE 2 (residues 1 to 141)
40 AUTHORS Brunel,D., Froger,N. and Pelletier,G.
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TITLE Direct Submission
JOURNAL Submitted (O1-APR-1998) Station de Genetique et
d'Amelioration des Plantes, INRA, Route de St Cyr,
Versailles
78026, France
$ COMMENT Method: conceptual translation.
FEATURES Location/Qualifiers
source 1..141
/organism="Brassica oleracea"
/cultivar="Rapide Cycling"
l~ /db xref="taxon:3712"
Protei n <1..>141
/product="unknown"
CDS 1..141
/partial
1$ /gene="FAD31"
/note="similar to Arabidopsis thaliana FADS"
coded by="join(AF056571.1:<235..327,AF056571.
1:436..621, AF056571.1:699..779,
AF056571.1:865..>927)"
ZO ORIGIN (SEQID N0: 41)
lpeklyknls hstrmlrytv plpmlayply lwyrspgkeg shynpysslf apserkliat
sttcwsivla tlvylsflvg pvtvlkvygv pyiifvmwld avtylhhhgh ddklpwyrgk
121 ewsylrgglt tvdrdygifn n
2$ AAD41580
(Brassica
napus)
LOCUS AF056570 1 141 as PLN O1-JUL-1999
DEFINITION unknown.
ACCESSION AAD41580
PID 85305310
3~ VERSION AAD41580.1 GI:5305310
DBSOURCE locus AF056570 accession AF056570.1
KEYWORDS
SOURCE rape.
ORGANISM Brassica napus
3$ Eukaryota; Viridiplantae; Streptophyta; Embryophyta;
Tracheophyta; euphyllophytes; Spermatophyta; Magnoliophyta;
eudicotyledons; core eudicots; Rosidae; eurosids
II;
Brassicales; Brassicaceae; Brassica.
REFERENCE 1 (residues 1 to 141)
~ AUTHORS Brunel,D., Froger,N. and Pelletier,G.
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TITLE Development of amplified consensus genetic markers
(A.C.G.M.)
I in Brassica napus from Arabidopsis thaliana sequences
of
known biological function
JOURNAL Unpublished
REFERENCE 2 (residues 1 to 141)
AUTHORS Brunel,D., Froger,N, and Pelletier,G.
TITLE Direct Submission
JOURNAL Submitted (O1-APR-1998) Station de Genetique et
d'Amelioration des Plantes, INRA, Route de St Cyr,
Versailles 78026, France
COMMENT Method: conceptual translation.
FEATURES Location/Qualifiers
source 1..141
/organism="Brassica napus"
/cultivar="Darmor"
/db xref="taxon:3708"
Protei n <1..>141
/product="unknown"
CDS 1..141
/partial
/gene="FAD32"
/note="similar to Arabidopsis thaliana FADS"
/coded by="join(AF056570.1:<107..199,AF056570.1:308.
.493,
2$ AF056570.1:572..652,AF056570.1:738..>800)"
ORIGIN ID N0: 42)
(SEQ
lpeklyknlshstrmlrytv plpmlayply lwyrspgkeg shynpysslf apserkliat
sttcwsivlaslvylsflvg pvtvlkvygv pyiifvmwld avtylhhhgh ddklpwyrgk
ewsylrgglttvdrdygifn n
Example 3
Cloning of the Fad3A gene by PCR from the 'A' genome of B. napus Apollo also
amplified fragments of the 'C' genome which represent a second FAD3 gene,
designated
Fad3C herein, from the 'C' genome of the low linolenic acid B. napus variety
Apollo.
Sequence polymorphisms in the Fad3C sequence have been identified that
facilitate
mapping the Fad3C gene. A partial genomic DNA sequence of Fad3C is shown in
Figure
1 l, a partial cDNA sequence of Fad3C is shown in Figure 7 and a partial amino
acid
seqence of Fad3C is shown in Figure 2.
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Conclusion
Although various embodiments of the invention are disclosed herein, many
adaptations and modifications may be made within the scope of the invention in
accordance with the common general knowledge of those skilled in this art.
Such
modifications include the substitution of known equivalents for any aspect of
the invention
in order to achieve the same result in substantially the same way. Numeric
ranges are
inclusive of the numbers defining the range. All documents referred to herein
are hereby
incorporated by reference, although no admission is made that any such
documents
constitute prior art. In the claims, the word "comprising" is used as an open-
ended term,
substantially equivalent to the phrase "including, but not limited to".
