Methods for increasing secretion of overexpressed proteins

The present invention is directed to methods for increasing secretion of an overexpressed gene product present in a host cell, by inducing expression of chaperone proteins within the host cell.

FIELD OF THE INVENTION 
The present invention relates to methods for increasing protein secretion 
of overexpressed gene products by enhancing chaperone protein expression 
within a host cell. Chaperone proteins which can increase protein 
secretion include protein folding chaperone proteins which bind to and 
assist in the folding of unfolded polypeptides. Such protein folding 
chaperone proteins include heat shock protein 70 (hsp70) class of proteins 
such as mammalian or yeast HSP68, HSP70, HSP72, HSP73, clathrin uncoating 
ATPase, IgG heavy chain binding protein (BiP), glucose-regulated proteins 
75, 78 and 80 (GRP75, GRP78 and GRP80), HSC70, and yeast KAR2, BiP, 
SSA1-4, SSB1, SSD1 and the like. Chaperone proteins which can increase 
protein secretion also include enzymes which catalyze covalent 
modification of proteins, such as mammalian or yeast protein disulfide 
isomerase (PDI), prolyl-4-hydroxylase .beta.-subunit, ERp59, glycosylation 
site binding protein (GSBP) and thyroid hormone binding protein (T3BP). 
BACKGROUND OF THE INVENTION 
Many proteins can be reversibly unfolded and refolded in vitro at dilute 
concentrations since all of the information required to specify a compact 
folded protein structure is present in the amino acid sequence of a 
protein. However, protein folding in vivo occurs in a concentrated milieu 
of numerous proteins in which intermolecular aggregation reactions compete 
with the intramolecular folding process. 
Moreover, gene products which are highly overexpressed are often poorly 
secreted even though secretion signals are present on such overexpressed 
gene products (Biemans et al. 1991 DNA Cell Biol. 10: 191-200; Elliot et 
al. 1989 Gene 79: 167-180; and Moir et al. 1987 Gene 56: 209-217). The 
prior art has not provided a clear reason for, or a simple and efficient 
means to overcome, such poor secretion of overexpressed gene products. 
Recently, a class of proteins have been identified which are associated 
with the intracellular folding of nascently formed polypeptides. Such 
proteins have been named `chaperone` proteins (e.g. see reviews by Ellis 
et al. 1991 Annu. Rev. Biochem. 60: 321-347; Gething et al. (1992) Nature 
355: 33-45; Rothman 1989 Cell 59: 591-601; Horwich et al. 1990 TIBTECH 8: 
126-131; and Morimoto et al. (Eds.) 1990 Stress Proteins in Biology and 
Medicine, Cold Spring Harbor Press: Cold Spring Harbor, N.Y., pp. 1-450). 
At least two classes of chaperone proteins are involved in polypeptide 
folding in cells. Enzymes such as protein disulfide isomerase (PDI) and 
peptidyl prolyl isomerase (PPI) can covalently modify proteins by 
catalyzing specific isomerization steps that may limit the folding rate of 
some proteins. (Freedman, R. B. 1989 Cell 57: 1067-1072). Another type of 
chaperone binds to folding intermediates but not to folded proteins and 
apparently causes no covalent modification of such intermediates. This 
latter type is referred to herein as a protein folding chaperone. 
Chaperone proteins that can covalently modify proteins include PDI and PPI. 
PDI catalyzes thiol/disulfide interchange reactions and promotes disulfide 
formation, isomerization or reduction, thereby facilitating the formation 
of the correct disulfide pairings, and may have a more general role in the 
prevention of premature misfolding of newly translocated chains. 
PDI interacts directly with newly synthesized secretory proteins and is 
required for the folding of nascent polypeptides in the endoplasmic 
reticulum (ER) of eukaryotic cells. Enzymes found in the ER with PDI 
activity include mammalian PDI (Edman et al., 1985, Nature 317:267), yeast 
PDI (Mizunaga et al. 1990, J. Biochem. 108:848), mammalian ERp59 
(Mazzarella et al., 1990, J. Biochem. 265:1094), mammalian 
prolyl-4-hydroxylase (Pihlajaniemi et al., 1987, EMBO J. 6: 643) yeast 
GSBP (Lamantia et al., 1991, Proc. Natl. Acad. Sci. USA, 88:4453) and 
mammalian T3BP (Yamauchi et al., 1987, Biochem. Biophys. Res. Commun. 
146:1485), and yeast EUG1 (Tachibana et al., 1992, Mol. Cell Biol. 12, 
4601). 
Two major families of protein folding chaperones have been identified, a 
heat shock protein 60 (hsp60) class and a heat shock protein 70 (hsp70) 
class. Chaperones of the hsp60 class are structurally distinct from 
chaperones of the hsp70 class. In particular, hsp60 chaperones appear to 
form a stable scaffold of two heptamer rings stacked one atop another 
which interacts with partially folded elements of secondary structure 
(Ellis et al. 1991; and Landry et al. 1992 Nature 355: 455-457). On the 
other hand, hsp70 chaperones are monomers or dimers and appear to interact 
with short extended regions of a polypeptide (Freiden et al. 1992 EMBO J. 
11: 63-70; and Landry et al. 1992). Hsp70 and hsp60 chaperones may also 
have sequential and complementary protein folding roles wherein hsp70 
proteins bind to extended polypeptide chains to prevent aggregation and 
hsp60 oligomers complete the folding of the extended polypeptide chain 
(Langer et al. 1992 Nature 354: 683-689). 
While hsp60 homologs appear to exist mainly within mitochondria and 
chloroplasts of eukaryotic cells, most compartments of eukaryotic cells 
contain members of the hsp70 class of chaperones. A eukaryotic hsp70 
homolog originally identified as the IgG heavy chain binding protein (BiP) 
is now known to have a more general role in associating with misfolded, 
unassembled or aberrantly glycosylated proteins. BiP is located in all 
eukaryotic cells within the lumen of the endoplasmic reticulum (ER). BiP 
is a soluble protein which is retained in the ER by a receptor-mediated 
recycling pathway and perhaps by calcium crosslinking (Pelham 1989 Annu. 
Rev. Cell. Biol. 5: 1-23; Sambrook 1990 Cell 61: 197-199). 
Hsp70 chaperones are well conserved in sequence and function (Morimoto et 
al. 1990). For example, the DnaK hsp70 protein chaperone in Escherichia 
coli, shares about 50% sequence homology with an hsp70 KAR2 chaperone in 
yeast (Rose et al. 1989 Cell 57:1211-1221). Moreover, the presence of 
mouse BiP in yeast can functionally replace a lost yeast KAR2 gene 
(Normington et al. 19: 1223-1236). Such a high structural and functional 
conservation for BiP has led to a generic usage for the term BiP as 
meaning any protein folding chaperone which resides in the endoplasmic 
reticulum of eukaryotes ranging from yeast to humans. 
The first step in the eukaryotic secretory pathway is translocation of the 
nascent polypeptide across the ER membrane in extended form. Correct 
folding and assembly of a polypeptide occurs in the ER and is a 
prerequisite for transport from the ER through the secretory pathway 
(Pelham 1989 Annu. Rev. Cell. Biol. 5: 1-23; Gething et al. 1990 Curr. Op. 
Cell Biol. 1: 65-72). For example, translocation intermediates which are 
artificially lodged in microsomal membranes in vitro can be chemically 
crosslinked with BiP (Sanders et al. 1992 Cell 69: 354-365). Therefore, 
misfolded proteins are retained in the ER, often in association with BiP 
(Suzuki et al. 1991 J. Cell Biol. 114: 189-205). 
The association of chaperone proteins with misfolded proteins has led some 
workers to conclude that hsp70 chaperone proteins like BiP act as 
proofreading proteins, whose chief role is to bind to and prevent 
secretion of misfolded proteins (Dorner et al. 1988 J. Mol. & Cell. Biol. 
8:4063-4070; Dorner et al. 1992 EMBO J. 11: 1563-1571). Dorner et al. 
(1992) have also suggested that overexpression of the BiP hsp70 chaperone 
protein can actually block secretion of selected proteins in Chinese 
hamster ovary cells. Therefore, according to the prior art, the role of 
BiP is to inhibit protein secretion. 
In contrast, the present invention provides methods for increasing protein 
secretion, unexpectedly, by increasing expression of an hsp70 chaperone 
protein or a PDI chaperone protein. Moreover, according to the present 
invention, it has been discovered that soluble forms of PDI and hsp70 
chaperone protein are diminished in cells which have been caused to 
overexpress a gene product. Therefore, the present methods can be used for 
increasing protein secretion by circumventing this dimunition of PDI 
and/or hsp70 chaperone protein expression. 
SUMMARY OF THE INVENTION 
The present invention provides a method for increasing secretion of 
overexpressed gene products from a host cell, which comprises expressing 
at least one chaperone protein in the host cell. In the present context, 
an overexpressed gene product is one which is expressed at levels greater 
than normal endogenous expression for that gene product. Overexpression 
can be effected, for example, by introduction of a recombinant 
construction that directs expression of a gene product in a host cell, or 
by altering basal levels of expression of an endogenous gene product, for 
example, by inducing its transcription. 
In one embodiment, the method of the invention comprises effecting the 
expression of at least one chaperone protein and an overexpressed gene 
product in a host cell, and cultivating said host cell under conditions 
suitable for secretion of the overexpressed gene product. The expression 
of the chaperone protein and the overexpressed gene product can be 
effected by inducing expression of a nucleic acid encoding the chaperone 
protein and a nucleic acid encoding the overexpressed gene product wherein 
said nucleic acids are present in a host cell. In another embodiment, the 
expression of the chaperone protein and the overexpressed gene product are 
effected by introducing a first nucleic acid encoding a chaperone protein 
and a second nucleic acid encoding a gene product to be overexpressed into 
a host cell under conditions suitable for expression of the first and 
second nucleic acids. In a preferred embodiment, one or both of said first 
and second nucleic acids are present in expression vectors. 
In another embodiment, expression of said chaperone protein is effected by 
inducing expression of a nucleic acid encoding said chaperone protein 
wherein said nucleic acid is present in a host cell or by introducing a 
nucleic acid encoding said chaperone protein into a host cell. Expression 
of said second protein is effected by inducing expression of a nucleic 
acid encoding said gene product to be overexpressed wherein said nucleic 
acid is present in a host cell or by introducing a nucleic acid encoding 
said second gene product into the host cell. 
In a preferred embodiment, the host cell is a yeast cell or a mammalian 
cell. 
In another preferred embodiment, the chaperone protein is an hsp70 
chaperone protein or a protein disulfide isomerase. The hsp70 chaperone 
protein is preferably yeast KAR2 or mammalian BiP. The protein disulfide 
isomerase is preferably yeast PDI or mammalian PDI. 
The present invention further provides a method for increasing secretion of 
an overexpressed gene product in a yeast host cell by using a yeast KAR2 
chaperone protein, or yeast PDI, or yeast KAR2 in combination with yeast 
PDI, in the present methods. 
The present invention also provides a method for increasing secretion of an 
overexpressed gene product in a mammalian host cell by using a mammalian 
BiP chaperone protein, or mammalian PDI, or mammalian BiP in combination 
with mammalian PDI, in the present methods.

DETAILED DESCRIPTION OF THE INVENTION 
According to the present invention, it has been discovered that the amount 
of chaperone proteins can be diminished in cells during overexpression of 
a gene product and this diminution in chaperone protein levels can lead to 
depressed protein secretion. Moreover, in accordance with the present 
invention it has been found that an increase in chaperone protein 
expression can increase secretion of an overexpressed gene product. 
Therefore, the present invention relates to a method for increasing 
secretion of an overexpressed gene product present in a host cell, which 
includes expressing a chaperone protein in the host cell and thereby 
increasing secretion of the overexpressed gene product. 
The present invention also contemplates a method of increasing secretion of 
an overexpressed gene product from a host cell by expressing a chaperone 
protein encoded by an expression vector present in or provided to the host 
cell, thereby increasing the secretion of the overexpressed gene product. 
The present invention provides a method for increasing secretion of 
overexpressed gene products from a host cell, which comprises expressing 
at least one chaperone protein in the host cell. In the present context, 
an overexpressed gene product is one which is expressed at levels greater 
than normal endogenous expression for that gene product. Overexpression 
can be effected, for example, by introduction of a recombinant 
construction that directs expression of a gene product in a host cell, or 
by altering basal levels of expression of an endogenous gene product, for 
example, by inducing its transcription. 
In one embodiment, the method of the invention comprises effecting the 
expression of at least one chaperone protein and an overexpressed gene 
product in a host cell, and cultivating said host cell under conditions 
suitable for secretion of the overexpressed gene product. The expression 
of the chaperone protein and the overexpressed gene product can be 
effected by inducing expression of a nucleic acid encoding the chaperone 
protein and a nucleic acid encoding the overexpressed gene product wherein 
said nucleic acids are present in a host cell. 
In another embodiment, the expression of the chaperone protein and the 
overexpressed gene product are effected by introducing a first nucleic 
acid encoding a chaperone protein and a second nucleic acid encoding a 
gene product to be overexpressed into a host cell under conditions 
suitable for expression of the first and second nucleic acids. In a 
preferred embodiment, one or both of said first and second nucleic acids 
are present in expression vectors. 
In another embodiment, expression of said chaperone protein is effected by 
inducing expression of a nucleic acid encoding said chaperone protein 
wherein said nucleic acid is present in a host cell or by introducing a 
nucleic acid encoding said chaperone protein into a host cell. Expression 
of said second protein is effected by inducing expression of a nucleic 
acid encoding said gene product to be overexpressed wherein said nucleic 
acid is present in a host cell or by introducing a nucleic acid encoding 
said second gene product into the host cell. 
In a preferred embodiment, the host cell is a yeast cell or a mammalian 
cell. 
In another preferred embodiment, the chaperone protein is an hsp70 
chaperone protein or a protein disulfide isomerase. The hsp70 chaperone 
protein is preferably yeast KAR2 or mammalian BiP. The protein disulfide 
isomerase is preferably yeast PDI or mammalian PDI. 
The present invention further provides a method for increasing secretion of 
an overexpressed gene product in a yeast host cell by using a yeast KAR2 
chaperone protein, or yeast PDI, or yeast KAR2 in combination with yeast 
PDI, in the present methods. 
The present invention also provides a method for increasing secretion of an 
overexpressed gene product in a mammalian host cell by using a mammalian 
BiP chaperone protein, or mammalian PDI, or mammalian BiP in combination 
with mammalian PDI, in the present methods. 
Chaperone proteins of the present invention include any chaperone protein 
which can facilitate or increase the secretion of proteins. In particular, 
members of the protein disulfide isomerase and heat shock 70 (hsp70) 
families of proteins are contemplated. An uncapitalized "hsp70" is used 
herein to designate the heat shock protein 70 family of proteins which 
share structural and functional similarity and whose expression are 
generally induced by stress. To distinguish the hsp70 family of proteins 
from the single heat shock protein of a species which has a molecular 
weight of about 70,000, and which has an art-recognized name of heat shock 
protein-70, a capitalized HSP70 is used herein. Accordingly, each member 
of the hsp70 family of proteins from a given species has structural 
similarity to the HSP70 protein from that species. 
The present invention is directed to any chaperone protein having the 
capability to stimulate secretion of an overexpressed gene product. The 
members of the hsp70 family of proteins are known to be structurally 
homologous. Moreover, according to the present invention any hsp70 
chaperone protein having sufficient homology to the KAR2 polypeptide 
sequence can be used in the present methods to stimulate secretion of an 
overexpressed gene product. Members of the PDI family are also 
structurally homologous, and any PDI which can be used according to the 
present method is contemplated herein. In particular, mammalian and yeast 
PDI, prolyl-4-hydroxylase .beta.-subunit, ERp59, GSBP and T3BP and yeast 
EUG1 are contemplated. 
As used herein, homology between polypeptide sequences is the degree of 
colinear similarity or identity between amino acids in one polypeptide 
sequence with that in another polypeptide sequence. Hence, homology can 
sometimes be conveniently described by the percentage, i.e. proportion, of 
identical amino acids in the sequences of the two polypeptides. For the 
present invention sufficient homology means that a sufficient percentage 
of sequence identity exists between an hsp70 chaperone polypeptide 
sequence and the KAR2 polypeptide sequence of SEQ ID NO:2, or between a 
PDI protein and the yeast PDI polypeptide sequence of SEQ ID NO:18 or the 
mammalian PDI sequence of SEQ ID NO:20 to retain the requisite function of 
the chaperone protein, i.e. stimulation of secretion. 
Therefore a sufficient number, but not necessarily all, of the amino acids 
in the present hsp70 chaperone polypeptide sequences are identical to the 
KAR2 polypeptide sequence of SEQ ID NO:2, or the yeast PDI polypeptide 
sequence of SEQ ID NO:18 or the mammalian PDI polypeptide of SEQ ID NO:20. 
In particular, the degree of homology between an hsp70 chaperone protein 
of the present invention and the polypeptide sequence of SEQ ID NO:2 need 
not be 100% so long as the chaperone protein can stimulate a detectable 
amount of gene product secretion. However, it is preferred that the 
present hsp70 chaperone proteins have at least about 50% homology with the 
polypeptide sequence of SEQ ID NO:2. In an especially preferred embodiment 
sufficient homology is greater than 60% homology with the KAR2 polypeptide 
sequence of SEQ ID NO:2. Similarly, the degree of homology between a PDI 
chaperone protein and the polypeptide sequence or SEQ ID NO:18 or 20 need 
not be 100% so long as the chaperone protein can stimulate a detectable 
amount of a gene product secretion. At least about 50% homology is 
preferred. 
The number of positions which are necessary to provide sufficient homology 
to KAR2 or PDI to retain the ability to stimulate secretion can be 
assessed by standard procedures for testing whether a chaperone protein of 
a given sequence can stimulate secretion. 
Procedures for observing whether an overexpressed gene product is secreted 
are readily available to the skilled artisan. For example, Goeddel, D. V. 
(Ed.) 1990, Gene Expression Technology, Methods in Enzymology, Vol 185, 
Academic Press, and Sambrook et al. 1989, Molecular Cloning: A Laboratory 
Manual, Vols. 1-3, Cold Spring Harbor Press, N.Y., provide procedures for 
detecting secreted gene products. 
To secrete an overexpressed gene product the host cell is cultivated under 
conditions sufficient for secretion of the overexpressed gene product. 
Such conditions include temperature, nutrient and cell density conditions 
that permit secretion by the cell. Moreover, such conditions are 
conditions under which the cell can perform basic cellular functions of 
transcription, translation and passage of proteins from one cellular 
compartment to another and are known to the skilled artisan. 
Moreover, as is known to the skilled artisan a secreted gene product can be 
detected in the culture medium used to maintain or grow the present host 
cells. The culture medium can be separated from the host cells by known 
procedures, e.g. centrifugation or filtration. The overexpressed gene 
product can then be detected in the cell-free culture medium by taking 
advantage of known properties characteristic of the overexpressed gene 
product. Such properties can include the distinct immunological, enzymatic 
or physical properties of the overexpressed gene product. 
For example, if an overexpressed gene product has a unique enzyme activity 
an assay for that activity can be performed on the culture medium used by 
the host cells. Moreover, when antibodies reactive against a given 
overexpressed gene product are available, such antibodies can be used to 
detect the gene product in any known immunological assay (e.g. as in 
Harlowe, et al., 1988, Antibodies: A Laboratory Manual, Cold Spring Harbor 
Laboratory Press). 
The secreted gene product can also be detected using tests that distinguish 
proteins on the basis of characteristic physical properties such as 
molecular weight. To detect the physical properties of the gene product 
all proteins newly synthesized by the host cell can be labeled, e.g. with 
a radioisotope. Common radioisotopes which are used to label proteins 
synthesized within a host cell include tritium (.sup.3 H), carbon-14 
(.sup.14 C), sulfur-35 (.sup.35 S) and the like. For example, the host 
cell can be grown in .sup.35 S-methionine or .sup.35 S-cysteine medium, 
and a significant amount of the .sup.35 S label will be preferentially 
incorporated into any newly synthesized protein, including the 
overexpressed protein. The .sup.35 S containing culture medium is then 
removed and the cells are washed and placed in fresh non-radioactive 
culture medium. After the cells are maintained in the fresh medium for a 
time and under conditions sufficient to allow secretion of the .sup.35 S 
radiolabelled overexpressed protein, the culture medium is collected and 
separated from the host cells. The molecular weight of the secreted 
labeled protein in the culture medium can then be determined by known 
procedures, e.g. polyacrylamide gel electrophoresis. Such procedures are 
described in more detail within Sambrook et al. (1989, Molecular Cloning: 
A Laboratory Manual, Vols. 1-3, Cold Spring Harbor Press, N.Y.). 
Thus for the present invention, one of ordinary skill in the art can 
readily ascertain which chaperone proteins have sufficient homology to 
KAR2 or PDI to stimulate secretion of an overexpressed gene product. 
According to the present invention, hsp70 chaperone proteins include yeast 
KAR2, HSP70, BiP, SSA1-4, SSB1, SSC1 and SSD1 gene products and eukaryotic 
hsp70 proteins such as HSP68, HSP72, HSP73, HSC70, clathrin uncoating 
ATPase, IgG heavy chain binding protein (BiP), glucose-regulated proteins 
75, 78 and 80 (GRP75, GRP78 and GRP80) and the like. 
Preferred PDI chaperone proteins include yeast and mammalian PDI, mammalian 
ERp59, mammalian prolyl-4-hydroxylase B-subunit, yeast GSBP, yeast EUG1 
and mammalian T3BP. 
Preferred chaperone proteins of the present invention normally reside 
within the endoplasmic reticulum of the host cell. For example, chaperone 
proteins which are localized with the endoplasmic reticulum include KAR2, 
GRP78, BiP, PDI and similar proteins. 
Moreover, the polypeptide sequence for the present hsp70 chaperones 
preferably has at least 50% sequence homology with a yeast KAR2 
polypeptide sequence having SEQ ID NO:2. The hsp70 chaperone polypeptide 
sequences which have at least 50% sequence homology with SEQ ID NO:2 
include, for example, any yeast HSP70, BiP, SSD1 and any mammalian or 
avian GRP78, HSP70 or HSC70. 
Preferred hsp70 chaperone polypeptide sequences include, for example: 
Saccharomyces cerevisiae KAR2 having a nucleotide sequence corresponding to 
SEQ ID NO:1 and a polypeptide sequence corresponding to SEQ ID NO:2 (Rose 
et al. 1989 Cell 57: 1211-1221; Normington et al. 1989 Cell 57: 
1223-1236); 
Schizosaccharomyces pombe HSP70 having a nucleotide sequence corresponding 
to SEQ ID NO:3 and a polypeptide sequence corresponding to SEQ ID NO:4 
(Powell et al. 1990 Gene 95:105-110); 
Kluyveromyces lactis BiP having a polypeptide sequence corresponding to SEQ 
ID NO:5 (Lewis et al. 1990 Nucleic Acids Res. 18: 6438); 
Schizosaccharomyces pombe BiP having a nucleotide sequence corresponding to 
SEQ ID NO:6 and a polypeptide sequence corresponding to SEQ ID NO:7 
(Pidoux et al. 1992 EMBO J. 11: 1583-1591); 
Saccharomyces cerevisiae SSD1 having a nucleotide sequence corresponding to 
SEQ ID NO:8 and a polypeptide sequence corresponding to SEQ ID NO:9 
(Sutton et al. 1991 Mol. Cell. Biol. 11: 2133-2148); 
Mouse GRP78 having a polypeptide sequence corresponding to SEQ ID NO:10; 
Hamster GRP78 having a polypeptide sequence corresponding to SEQ ID NO:11; 
Human GRP78 having a nucleotide sequence corresponding to SEQ ID NO:12 
(Ting et al. 1988 DNA 7: 275-286); 
Mouse HSC70 having a nucleotide sequence corresponding to SEQ ID NO:13 and 
a polypeptide sequence corresponding to SEQ ID NO:14 (Giebel et al. 1988 
Dev. Biol. 125: 200-207); 
Human HSC70 having a nucleotide sequence corresponding to SEQ ID NO:15 
(Dworniczak et al. 1987 Nucleic Acids Res. 15: 5181-5197); 
Chicken GRP78 having a polypeptide sequence corresponding to SEQ ID NO:16; 
Rat GRP78 as in Chang et al. (1987 Proc. Natl. Acad. Sci. USA 84: 680-684); 
Saccharomyces cerevisiae SCC-1 as in Craig et al. (1987 Proc. Natl. Acad. 
Sci. USA 84: 680-684); 
Preferred hsp70 proteins of the present invention are normally present in 
the endoplasmic reticulum of the cell. Preferred hsp70 proteins also 
include yeast KAR2, BiP, and HSP70 proteins, avian BiP or GRP78 proteins 
and mammalian BiP or GRP78 proteins. 
The polypeptide sequence for the present PDI chaperones preferably has at 
least 50% homology with the yeast PDI of SEQ ID NO:18 or the rat PDI of 
SEQ ID NO:20. Preferred PDI chaperone polypeptides include, for example, 
Saccharomyces cerevisiae PDI having a nucleotide sequence corresponding to 
SEQ ID NO:17 and a polypeptide sequence corresponding to SEQ ID NO:18 (La 
Mantia et al., 1991, Proc. Natl. Acad. Sci. USA 88: 4453-4457). 
Rat PDI having a nucleotide sequence corresponding to SEQ ID NO:19 and a 
polypeptide sequence corresponding to SEQ ID NO:20 (Edman et al., 1985 
Nature, 317:267). 
Human prolyl 4-hydroxylase .beta.-subunit having a nucleotide and amino 
acid sequence as disclosed by Pihlajaniemi et al., 1987, EMBO, J. 6: 
643-649. 
Bovine T3BP having a nucleotide and amino acid sequence as disclosed by 
Yamauchi et al, 1987, Biochem. Biophys. Res. Commun., 146:1485-1492. 
Murine ERp59 having a nucleotide and amino acid sequence as disclosed by 
Mazzarella et al., 1990, J. Biol. Chem. 265: 1094-1101. 
As is known to the skilled artisan, a given amino acid is encoded by 
different three-nucleotide codons. Such degeneracy in the genetic code 
therefore means that the same polypeptide sequence can be encoded by 
numerous nucleotide sequences. The present invention is directed to 
methods utilizing any nucleotide sequence which can encode the present 
hsp70 chaperone polypeptides. Therefore, for example, while the KAR2 
polypeptide sequence of SEQ ID NO:2 can be encoded by a nucleic acid 
comprising SEQ ID NO:1 there are alternative nucleic acid sequences which 
can encode the same KAR2 SEQ ID NO:2 polypeptide sequence. The present 
invention is also directed to use of such alternative nucleic acid 
sequences in the present methods. 
Moreover when the host cell is a yeast host cell the chaperone protein is 
preferably a yeast KAR2 or BiP protein or PDI protein, e.g. SEQ ID NO:2, 
SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:18 and homologues thereof. Accordingly 
the present invention also provides a method for increasing secretion of 
an overexpressed gene product present in or provided to a yeast host cell, 
which includes expressing at least one KAR2 or BiP or PDI chaperone 
protein in the host cell and thereby increasing secretion of the gene 
product. In one embodiment such a method can also include expressing at 
least one of a KAR2 or BiP or PDI chaperone protein encoded by at least 
one expression vector present in or provided to the host cell, and thereby 
increasing secretion of the overexpressed recombinant gene product. Such 
an expression vector can include a nucleic acid encoding a polypeptide 
sequence for a yeast KAR2 or BiP or PDI chaperone protein operably linked 
to a nucleic acid which effects expression of the yeast KAR2 or BiP or PDI 
chaperone protein. 
Yeast as used herein includes such species as Saccharomyces cerevisiae, 
Hansenula polymorpha, Kluyveromyces lactis, Pichia pastoris, 
Schizosaccharomyces pombe, Yarrowia lipolytica and the like. 
Furthermore, when an avian or mammalian host is used a BiP or GRP78 or 
mammalian PDI chaperone protein is preferably employed, e.g. any one of 
SEQ ID NO: 10-12, 16 or 20 and homologues thereof. Therefore, the present 
invention also provides a method for increasing secretion of an 
overexpressed gene product in a mammalian host cell, which includes 
expressing at least one of a BiP or GRP78 or mammalian PDI chaperone 
protein in the host cell and thereby increasing secretion of the gene 
product. Such a method can also include expressing a BiP or GRP78 or 
mammalian PDI chaperone protein encoded by an expression vector present in 
or provided to the host cell and thereby increasing the secretion of the 
overexpressed gene product. Such an expression vector can include a 
nucleic acid encoding a polypeptide sequence for the BiP or the GRP78 or 
the mammalian PDI chaperone protein operably linked to a sequence which 
effects expression of such a chaperone protein. 
In a preferred embodiment the chaperone protein is a mammalian or avian 
GRP78 protein, or a mammalian PDI. 
Mammals as used herein includes mouse, hamster, rat, monkey, human and the 
like. 
The present invention provides methods for increasing secretion of any 
overexpressed gene product which naturally has a secretion signal or has 
been genetically engineered to have a secretion signal. 
Secretion signals are discrete amino acid sequences which cause the host 
cell to direct a gene product through internal and external cellular 
membranes and into the extracellular environment. 
Secretion signals are present at the N-terminus of a nascent polypeptide 
gene product targeted for secretion. Additional eukaryotic secretion 
signals can also be present along the polypeptide chain of the gene 
product in the form of carbohydrates attached to specific amino acids, 
i.e. glycosylation secretion signals. 
N-terminal signal sequences include a hydrophobic domain of about 10 to 
about 30 amino acids which can be preceded by a short charged domain of 
about 2 to about 10 amino acids. Moreover, the signal sequence is present 
at the N-terminus of gene products destined for secretion. In general, the 
particular sequence of a signal sequence is not critical but signal 
sequences are rich in hydrophobic amino acids such as alanine (Ala), 
valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), 
phenylalanine (Phe), tryptophan (Trp), methionine (Met) and the like. 
Many signal sequences are known (Michaelis et al. 1982 Ann. Rev. Microbiol. 
36: 425). For example, the yeast acid phosphatase, yeast invertase and the 
yeast .alpha.-factor signal sequences have been attached to heterologous 
polypeptide coding regions and used successfully for secretion of the 
heterologous polypeptide (Sato et al. 1989 Gene 83: 355-365; Chang et al. 
1986 Mol. Cell. Biol. 6: 1812-1819; and Brake et al. 1984 Proc. Natl. 
Acad. Sci. USA 81: 4642-4646). Therefore, the skilled artisan can readily 
design or obtain a nucleic acid which encodes a coding region for an 
overexpressed gene product which also has a signal sequence at the 5'-end. 
Eukaryotic glycosylation signals include specific types of carbohydrates 
which are attached to specific types of amino acids present in a gene 
product. Carbohydrates which are attached to such amino acids include 
straight or branched chains containing glucose, fucose, mannose, 
galactose, N-acetylglucosamine, N-acetylgalactosamine, N-acetylneuraminic 
acid and the like. Amino acids which are frequently glycosylated include 
asparagine (Asn), serine (Ser), threonine (Thr), hydroxylysine and the 
like. 
Examples of overexpressed gene products which are preferably secreted by 
the present methods include mammalian gene products such as enzymes, 
cytokines, growth factors, hormones, vaccines, antibodies and the like. 
More particularly, preferred overexpressed gene products of the present 
invention include gene products such as erythropoietin, insulin, 
somatotropin, growth hormone releasing factor, platelet derived growth 
factor, epidermal growth factor, transforming growth factor .alpha., 
transforming growth factor .beta., epidermal growth factor, fibroblast 
growth factor, nerve growth factor, insulin-like growth factor I, 
insulin-like growth factor II, clotting Factor VIII, superoxide dismutase, 
.alpha.-interferon, .gamma.-interferon, interleukin-1, interleukin-2, 
interleukin-3, interleukin-4, interleukin-5, interleukin-6, granulocyte 
colony stimulating factor, multi-lineage colony stimulating activity, 
granulocyte-macrophage stimulating factor, macrophage colony stimulating 
factor, T cell growth factor, lymphotoxin and the like. Preferred 
overexpressed gene products are human gene products. 
Moreover, the present methods can readily be adapted to enhance secretion 
of any overexpressed gene product which can be used as a vaccine. 
Overexpressed gene products which can be used as vaccines include any 
structural, membrane-associated, membrane-bound or secreted gene product 
of a mammalian pathogen. Mammalian pathogens include viruses, bacteria, 
single-celled or multi-celled parasites which can infect or attack a 
mammal. For example, viral vaccines can include vaccines against viruses 
such as human immunodeficiency virus (HIV), R. rickettsii, vaccinia, 
Shigella, poliovirus, adenovirus, influenza, hepatitis A, hepatitis B, 
dengue virus, Japanese B encephalitis, Varicella zoster, cytomegalovirus, 
hepatitis A, rotavirus, as well as vaccines against viral diseases like 
Lyme disease, measles, yellow fever, mumps, rabies, herpes, influenza, 
parainfluenza and the like. Bacterial vaccines can include vaccines 
against bacteria such as Vibrio cholerae, Salmonella typhi, Bordetella 
pertussis, Streptococcus pneumoniae, Hemophilus influenza, Clostridium 
tetani, Corynebacterium diphtheriae, Mycobacterium leprae, Neisseria 
gonorrhoeae, Neisseria meningitidis, Coccidioides immitis and the like. 
Moreover, an overexpressed gene product of the present invention can be 
overexpressed from its own natural promoter, from a mutated form of such a 
natural promoter or from a heterologous promoter which has been operably 
linked to a nucleic acid encoding the gene product. Accordingly, 
overexpressed gene products contemplated by the present invention include 
recombinant and non-recombinant gene products. As used herein a 
recombinant gene product is a gene product expressed from a nucleic acid 
which has been isolated from the natural source of such a gene product or 
nucleic acid. In contrast, non-recombinant, or native, gene products are 
expressed from nucleic acids naturally present in the host cell. 
Therefore, the present overexpressed gene products can be native products 
of the host cell which are naturally produced at high levels, e.g. 
antibodies, enzymes, cytokines, hormones and the like. Moreover, if the 
factors controlling expression of a native gene product are understood, 
such factors can also be manipulated to achieve overexpression of the gene 
product, e.g. by induction of transcription from the natural promoter 
using known inducer molecules, by mutation of the nucleic acids 
controlling or repressing expression of the gene product to produce a 
mutant strain that constitutively overexpresses the gene product, by 
second site mutations which depress the synthesis or function of factors 
which normally repress the transcription of the gene product, and the 
like. 
Similarly, the present chaperone proteins can be expressed 
non-recombinantly, i.e. from the host cell's native gene for that 
chaperone protein, by manipulating the factors controlling expression of 
the native chaperone protein to permit increased expression of the 
chaperone protein. For example, the native hsp70 chaperone gene or the 
transcriptional or translational control elements for the hsp70 chaperone 
can be mutated so that the hsp70 chaperone protein is constitutively 
expressed. Alternatively, nucleic acids encoding factors which control the 
transcription or translation of the chaperone protein can be mutated to 
achieve increased expression of the chaperone protein. Such mutations can 
thereby overcome the decrease in native chaperone protein expression which 
occurs upon overexpression of a gene product. 
The overexpressed gene products and the chaperone proteins of the present 
invention can also be expressed recombinantly, i.e. by placing a nucleic 
acid encoding a gene product or a chaperone protein into an expression 
vector. Such an expression vector minimally contains a sequence which 
effects expression of the gene product or the chaperone protein when the 
sequence is operably linked to a nucleic acid encoding the gene product or 
the chaperone protein. Such an expression vector can also contain 
additional elements like origins of replication, selectable markers, 
transcription or termination signals, centromeres, autonomous replication 
sequences, and the like. 
According to the present invention, first and second nucleic acids encoding 
an overexpressed gene product and a chaperone protein, respectively, can 
be placed within expression vectors to permit regulated expression of the 
overexpressed gene product and/or the chaperone protein. While the 
chaperone protein and the overexpressed gene product can be encoded in the 
same expression vector, the chaperone protein is preferably encoded in an 
expression vector which is separate from the vector encoding the 
overexpressed gene product. Placement of nucleic acids encoding the 
chaperone protein and the overexpressed gene product in separate 
expression vectors can increase the amount of secreted overexpressed gene 
product. 
As used herein, an expression vector can be a replicable or a 
non-replicable expression vector. A replicable expression vector can 
replicate either independently of host cell chromosomal DNA or because 
such a vector has integrated into host cell chromosomal DNA. Upon 
integration into host cell chromosomal DNA such an expression vector can 
lose some structural elements but retains the nucleic acid encoding the 
gene product or the hsp70 chaperone protein and a segment which can effect 
expression of the gene product or the chaperone protein. Therefore, the 
expression vectors of the present invention can be chromosomally 
integrating or chromosomally nonintegrating expression vectors. 
In a preferred embodiment of the present invention, one or more chaperone 
proteins are overexpressed in a host cell by introduction of integrating 
or nonintegrating expression vectors into the host cell. Following 
introduction of at least one expression vector encoding at least one 
chaperone protein, the gene product is then overexpressed by inducing 
expression of an endogenous gene encoding the gene product, or by 
introducing into the host cell an expression vector encoding the gene 
product. In another preferred embodiment, cell lines are established which 
constitutively or inducibly express at least one chaperone protein. An 
expression vector encoding the gene product to be overexpressed is 
introduced into such cell lines to achieve increased secretion of the 
overexpressed gene product. 
The present expression vectors can be replicable in one host cell type, 
e.g., Escherichia coli, and undergo little or no replication in another 
host cell type, e.g., a eukaryotic host cell, so long as an expression 
vector permits expression of the present chaperone proteins or 
overexpressed gene products and thereby facilitates secretion of such gene 
products in a selected host cell type. 
Expression vectors as described herein include DNA or RNA molecules 
engineered for controlled expression of a desired gene, i.e. a gene 
encoding the present chaperone proteins or a overexpressed gene product. 
Such vectors also encode nucleic acid segments which are operably linked 
to nucleic acids encoding the present chaperone polypeptides or the 
present overexpressed gene products. Operably linked in this context means 
that such segments can effect expression of nucleic acids encoding 
chaperone protein or overexpressed gene products. These nucleic acid 
sequences include promoters, enhancers, upstream control elements, 
transcription factors or repressor binding sites, termination signals and 
other elements which can control gene expression in the contemplated host 
cell. Preferably the vectors are plasmids, bacteriophages, cosmids or 
viruses. 
Sambrook et al. 1989; Goeddel, 1990; Perbal, B. 1988, A Practical Guide to 
Molecular Cloning, John Wiley & Sons, Inc.; and Romanos et al. 1992, Yeast 
8: 423-488, provide detailed reviews of vectors into which a nucleic acid 
encoding the present chaperone polypeptide sequences or the contemplated 
overexpressed gene products can be inserted and expressed. 
Expression vectors of the present invention function in yeast or mammalian 
cells. Yeast vectors can include the yeast 2.mu. circle and derivatives 
thereof, yeast plasmids encoding yeast autonomous replication sequences, 
yeast minichromosomes, any yeast integrating vector and the like. A 
comprehensive listing of many types of yeast vectors is provided in Parent 
et al. (1985 Yeast 1: 83-138). Mammalian vectors can include SV40 based 
vectors, polyoma based vectors, retrovirus based vectors, Epstein-Barr 
virus based vectors, papovavirus based vectors, bovine papilloma virus 
(BPV) vectors, vaccinia virus vectors, baculovirus vectors and the like. 
Muzyczka (ed. 1992 Curr. Top. Microbiol. Immunol. 158:97-129) provides a 
comprehensive review of eukaryotic expression vectors. 
Elements or nucleic acid sequences capable of effecting expression of a 
gene product include promoters, enhancer elements, upstream activating 
sequences, transcription termination signals and polyadenylation sites. 
All such promoter and transcriptional regulatory elements, singly or in 
combination, are contemplated for use in the present expression vectors. 
Moreover, genetically-engineered and mutated regulatory sequences are also 
contemplated herein. 
Promoters are DNA sequence elements for controlling gene expression. In 
particular, promoters specify transcription initiation sites and can 
include a TATA box and upstream promoter elements. 
Yeast promoters are used in the present expression vectors when a yeast 
host cell is used. Such yeast promoters include the GAL1, PGK, GAP, TPI, 
CYC1, ADH2, PHO5, CUP1, MF.alpha.1, MF.alpha.1 and related promoters. 
Romanos et al. (1992 Yeast 8: 423-488) provide a review of yeast promoters 
and expression vectors. 
Higher eukaryotic promoters which are useful in the present expression 
vectors include promoters of viral origin, such as the baculovirus 
polyhedrin promoter, the vaccinia virus hemagglutinin (HA) promoter, SV40 
early and late promoter, the herpes simplex thymidine kinase promoter, the 
Rous sarcoma virus LTR, the Moloney Leukemia Virus LTR, and the Murine 
Sarcoma Virus (MSV) LTR. Sambrook et al. (1989) and Goeddel (1990) review 
higher eukaryote promoters. 
Preferred promoters of the present invention include inducible promoters, 
i.e. promoters which direct transcription at an increased or decreased 
rate upon binding of a transcription factor. Transcription factors as used 
herein include any factor that can bind to a regulatory or control region 
of a promoter an thereby affect transcription. The synthesis or the 
promoter binding ability of a transcription factor within the host cell 
can be controlled by exposing the host to an inducer or removing an 
inducer from the host cell medium. Accordingly to regulate expression of 
an inducible promoter, an inducer is added or removed from the growth 
medium of the host cell. Such inducers can include sugars, phosphate, 
alcohol, metal ions, hormones, heat, cold and the like. For example, 
commonly used inducers in yeast are glucose, galactose, and the like. 
The expression vectors of the present invention can also encode selectable 
markers. Selectable markers are genetic functions that confer an 
identifiable trait upon a host cell so that cells transformed with a 
vector carrying the selectable marker can be distinguished from 
non-transformed cells. Inclusion of a selectable marker into a vector can 
also be used to ensure that genetic functions linked to the marker are 
retained in the host cell population. Such selectable markers can confer 
any easily identified dominant trait, e.g. drug resistance, the ability to 
synthesize or metabolize cellular nutrients and the like. 
Yeast selectable markers include drug resistance markers and genetic 
functions which allow the yeast host cell to synthesize essential cellular 
nutrients, e.g. amino acids. Drug resistance markers which are commonly 
used in yeast include chloramphenicol (Cm.sup.r), kanamycin (kan.sup.r), 
methotrexate (mtx.sup.r or DHFR.sup.+) G418 (geneticin) and the like. 
Genetic functions which allow the yeast host cell to synthesize essential 
cellular nutrients are used with available yeast strains having 
auxotrophic mutations in the corresponding genomic function. Common yeast 
selectable markers provide genetic functions for synthesizing leucine 
(LEU2), tryptophan (TRP1), uracil (URA3), histidine (HIS3), lysine (LYS2) 
and the like. 
Higher eukaryotic selectable markers can include genetic functions encoding 
an enzyme required for synthesis of a required nutrient, e.g. the 
thymidine kinase (tk), dihydrofolate reductase (DHFR), uridine (CAD), 
adenosine deaminase (ADA), asparagine synthetase (AS) and the like. The 
presence of some of these enzymatic functions can also be identified by 
exposing the host cell to a toxin which can be inactivated by the enzyme 
encoded by the selectable marker. Moreover drug resistance markers are 
available for higher eukaryotic host cells, e.g. aminoglycoside 
phosphotransferase (APH) markers are frequently used to confer resistance 
to kanamycin, neomycin and geneticin, and hygromycin B phosphotransferase 
(hyg) confers resistance to hygromycin in higher eukaryotes. Some of the 
foregoing selectable markers can also be used to amplify linked genetic 
functions by slowly adding the appropriate substrate for the enzyme 
encoded by markers such as DHFR, CAD, ADA, AS and others. 
Therefore the present expression vectors can encode selectable markers 
which are useful for identifying and maintaining vector-containing host 
cells within a cell population present in culture. In some circumstances 
selectable markers can also be used to amplify the copy number of the 
expression vector. 
After inducing transcription from the present expression vectors to produce 
an RNA encoding an overexpressed gene product or a chaperone protein, the 
RNA is translated by cellular factors to produce the gene product or the 
chaperone protein. 
In yeast and other eukaryotes, translation of a messenger RNA (mRNA) is 
initiated by ribosomal binding to the 5' cap of the mRNA and migration of 
the ribosome along the mRNA to the first AUG start codon where polypeptide 
synthesis can begin. Expression in yeast and mammalian cells generally 
does not require specific number of nucleotides between a 
ribosomal-binding site and an initiation codon, as is sometimes required 
in prokaryotic expression systems. However, for expression in a yeast or a 
mammalian host cell, the first AUG codon in an mRNA is preferably the 
desired translational start codon. 
Moreover, when expression is performed in a yeast host cell the presence of 
long untranslated leader sequences, e.g. longer than 50-100 nucleotides, 
can diminish translation of an mRNA. Yeast mRNA leader sequences have an 
average length of about 50 nucleotides, are rich in adenine, have little 
secondary structure and almost always use the first AUG for initiation 
(Romanos et al. 1992; and Cigan et al. 1987 Gene 59: 1-18). Since leader 
sequences which do not have these characteristics can decrease the 
efficiency of protein translation, yeast leader sequences are preferably 
used for expression of an overexpressed gene product or a chaperone 
protein in a yeast host cell. The sequences of many yeast leader sequences 
are known and are available to the skilled artisan, e.g. by reference to 
Cigan et al. (1987 Gene 59: 1-18). 
In mammalian cells, nucleic acids encoding chaperone proteins or 
overexpressed gene products generally include the natural 
ribosomal-binding site and initiation codon because, while the number of 
nucleotides between transcription and translational start sites can vary, 
such variability does not greatly affect the expression of the polypeptide 
in a mammalian host. However, when expression is performed in a mammalian 
host cell, the first AUG codon in an mRNA is preferably the desired 
translational start codon. 
In addition to the promoter, the ribosomal-binding site and the position of 
the start codon, factors which can effect the level of expression obtained 
include the copy number of a replicable expression vector. The copy number 
of a vector is generally determined by the vector's origin of replication 
and any cis-acting control elements associated therewith. For example, an 
increase in copy number of a yeast episomal vector encoding a regulated 
centromere can be achieved by inducing transcription from a promoter which 
is closely juxtaposed to the centromere (Chlebowicz-Sledziewska et al. 
1985 Gene 39: 25-31). Moreover, encoding the yeast FLP function in a yeast 
vector can also increase the copy number of the vector (Romanos et al.). 
The skilled artisan has available many choices of expression vectors. For 
example, commonly available yeast expression vectors include pWYG-4, 
pWYG7L and the like. Goeddel (1990) provides a comprehensive listing of 
yeast expression vectors and sources for such vectors. Commercially 
available higher eukaryotic expression vectors include pSVL, PMSG, 
pKSV-10, pSVN9 and the like. 
One skilled in the art can also readily design and make expression vectors 
which include the above-described sequences by combining DNA fragments 
from available vectors, by synthesizing nucleic acids encoding such 
regulatory elements or by cloning and placing new regulatory elements into 
the present vectors. Methods for making expression vectors are well-known. 
Overexpressed DNA methods are found in any of the myriad of standard 
laboratory manuals on genetic engineering (Sambrook et al., 1989; Goeddel, 
1990 and Romanos et al. 1992). 
For example, a centromere-containing YCp50 vector (Goeddel, 1990) which 
encodes a URA3 selectable marker can be modified to encode an associated 
inverted sequence which permits high copy number replication in yeast. A 
galactose inducible promoter, e.g. PGAL1, can be placed within such a 
vector and a chaperone polypeptide sequence, e.g., SEQ ID NO:2 can be 
inserted immediately downstream. A pSC101 origin of replication can also 
be used in such a vector to permit replication at low copy numbers in 
Escherichia coli. One such replicable expression vector which has such 
structural elements is a pMR1341 vector (Vogel et al. 1990 J. Cell. Biol. 
110: 1885). 
The expression vectors of the present invention can be made by ligating the 
present chaperone protein coding regions in the proper orientation to the 
promoter and other sequence elements being used to control gene 
expression. This juxtapositioning of promoter and other sequence elements 
with the present hsp70 chaperone polypeptide coding regions allows 
synthesis of large amounts of the chaperone polypeptide which can then 
increase secretion of a co-synthesized overexpressed protein. 
After construction of the present expression vectors, such vectors are 
transformed into host cells where the overexpressed gene product and the 
chaperone protein can be expressed. Methods for transforming yeast and 
higher eukaryotic cells with expression vectors are well known and readily 
available to the skilled artisan. 
For example, expression vectors can be transformed into yeast cells by any 
of several procedures including lithium acetate, spheroplast, 
electroporation and similar procedures. Such procedures can be found in 
numerous references including Ito et al. (1983, J. Bacteriol. 153: 163), 
Hinnen et al. (1978 Proc. Natl. Acad. Sci. U.S.A. 75: 1929) and Guthrie et 
al. (1991 Guide to Yeast Genetics and Molecular Biology, in Methods In 
Enzymology, vol. 194, Academic Press, New York). 
Mammalian host cells can also be transformed with the present expression 
vectors by a variety of techniques including transfection, infection and 
other transformation procedures. For example, transformation procedures 
include calcium phosphate-mediated, DEAE-dextran-mediated or 
polybrene-mediated transformation, protoplast or liposomal fusion, 
electroporation, direct microinjection into nuclei and the like. Such 
procedures are provided in Sambrook et al. and the references cited 
therein. 
Yeast host cells which can be used with yeast replicable expression vectors 
include any wild type or mutant strain of yeast which is capable of 
secretion. Such strains can be derived from Saccharomyces cerevisiae, 
Hansenula polymorpha, Kluyveromyces lactis, Pichia pastoris, 
Schizosaccharomyces pombe, Yarrowia lipolytica and related species of 
yeast. In general, preferred mutant strains of yeast are strains which 
have a genetic deficiency that can be used in combination with a yeast 
vector encoding a selectable marker. Many types of yeast strains are 
available from the Yeast Genetics Stock Center (Donner Laboratory, 
University of California, Berkeley, Calif. 94720), the American Type 
Culture Collection (12301 Parklawn Drive, Rockville, Md. 20852, 
hereinafter ATCC), the National Collection of Yeast Cultures (Food 
Research Institute, Colney Lane, Norwich NR4 7UA, UK) and the 
Centraalbureau voor Schimmelcultures (Yeast Division, Julianalaan 67a, 
2628 BC Delft, Netherlands). 
Tissue culture cells that are used with eukaryotic expression vectors can 
include VERO cells, NRC-5 cells, SCV-1 cells, COS-1 cells, CV-1 cells, 
LCC-MK.sub.2 cells, NIH3T3 cells, CHO-K1 cells, mouse L cells, HeLa cells, 
Antheraea eucalypti moth ovarian cells, Aedes aegypti mosquito cells, S. 
frugiperda cells and other cultured cell lines known to one skilled in the 
art. Such host cells can be obtained from the ATCC. For example, Table 1 
provides examples of higher eukaryotic host cells which are illustrative 
of the many types of host cells which can be used with the present 
methods. The subject matter of Table 1 is not intended to limit the 
invention is any respect. 
The following Examples further illustrate the invention. 
TABLE 1 
______________________________________ 
HOST CELL ORIGIN SOURCE 
______________________________________ 
Aedes aegypti 
Mosquito Larvae *ATCC #CCL 125 
LtK- Mouse Exp. Cell. Res 
31:297-312 
CV-1 African Green Monkey 
ATCC #CCL 70 
Kidney 
LCC-MK.sub.2 original 
Rhesus Monkey Kidney 
ATCC #CCL 7 
LCC-MK.sub.2 derivative 
Rhesus Monkey Kidney 
ATCC #CCL 7.1 
3T3 Mouse Embryo Fibroblasts 
ATCC #CCL 92 
CHO-K1 Chinese Hamster Ovary 
ATCC #CCL 61 
293 Human Embryonic Kidney 
ATCC #CRL 1573 
Antheraea eucalypti 
Moth Ovarian Tissue 
ATCC #CCL 80 
HeLa Human Cervix Epitheloid 
ATCC #CCL 2 
C1271 Mouse Fibroblast 
ATCC #CRL 1616 
HS-Sultan Human Plasma Cell 
ATCC #CRL 1484 
Plasmacytoma 
Saccharomyces ATCC #44773 
cerevisiae DBY746 
______________________________________ 
*American Type Culture Collection, 1201 Parklawn Drive, Rockville, 
Maryland 
EXAMPLE 1 
Effect of Overexpression of Proteins on Native Yeast Chaperone Protein 
Synthesis 
The expression of native yeast chaperone KAR2 protein was observed in yeast 
cells constitutively overexpressing human gene products erythropoietin, 
granulocyte colony stimulating factor, platelet derived growth factor or 
Schizosaccharomyces pombe acid phosphatase. These non-yeast products have 
a variety of distinct structural features including different sizes, 
differences in glycosylation, and different numbers of subunits (Table 2). 
Materials and Methods: 
TABLE 2 
______________________________________ 
STRUCTURAL FEATURES OF OVEREXPRESSED 
GENE PRODUCTS 
Protein.sup.a 
Multiple Subunits? 
Glycosylated? 
Size (kd) 
______________________________________ 
EPO + 193 
PDGF + 241 
GCSF 207 
PHO + + 435 
GCSF-PHO + + 548 
______________________________________ 
.sup.a EPO = human erythropoietin, PDGF = human platelet derived growth 
factor B chain, GCSF = human granulocyte colony stimulating factor, PHO = 
Schizosaccharomyces pombe acid phosphatase, and GCSFPHO = fusion between 
GCSF and PHO. 
Yeast YPH500 (.alpha. ura3-52 lys2-801a ade2-101 trp-.DELTA.63 
his3-.DELTA.200 leu2-.DELTA.1) cells were transformed with multicopy 
plasmids encoding one of the overexpressed gene products described in 
Table 2, using methods provided in Guthrie et al. and then cultured in 
protein-free Synthetic Complete (SC) media. Extracts from 10 ml cultures 
of mid-exponential growing cells were prepared by glass bead disruption 
(Guthrie et al). Serial dilutions were made of protein extracts from 
strains expressing the different gene products. Equal amounts of total 
protein were loaded onto a BioRad slot blotting were prepared. 
The blots were probed with anti-KAR2 antibody followed by goat anti-rabbit 
secondary antibody conjugated to alkaline phosphatase. Alkaline 
phosphatase enzymatic activity was detected by use of a Lumi-Phos 
530.sup.R substrate (Boehringer Mannheim) to form a chemi-luminescent 
product. Quantitation of the amount of KAR2 protein expressed in different 
cell extracts was by densitometric scanning of X-ray films exposed to 
blots treated with Lumi-Phos 530.sup.R. 
Results: 
FIG. 1 depicts the amounts of KAR2 protein in wild type yeast and yeast 
strains which had been overexpressing human erythropoietin (EPO), human 
platelet derived growth factor B chain (PDGF), human granulocyte colony 
stimulating factor (GCSF), Schizosaccharomyces pombe acid phosphatase 
(PHO) and a fusion between GCSF and PHO (GCSF-PHO) for 50 or more 
generations. 
Surprisingly, native soluble KAR2 protein levels were at least five-fold 
lower in cells expressing these foreign genes from multicopy plasmids. 
Lower levels of expression from a single-copy control plasmid (i.e. 
single-copy PHO) did not greatly diminish KAR2 protein expression. 
Similar results were obtained when using a BJ5464 yeast strain (.alpha. 
ura3-52 trp1 leu2.DELTA.1 his3.DELTA.200 pep4::HIS3 prb1.DELTA.1.6R can1 
GAL), which is deficient in vacuolar proteases. Therefore, the differences 
in KAR2 expression were not due to differences in the levels of vacuolar 
proteases. Moreover, the addition of other protease inhibitors to the cell 
extracts did not change the relative amount of KAR2 protein observed. 
Further, mixing experiments of cellular extracts containing and not 
containing KAR2, confirmed that proteolysis during sample preparation was 
negligible. Therefore, strain-dependent differences in proteolysis could 
not account for the observed dimunition of KAR2 protein expression in 
yeast strains overexpressing proteins from multicopy plasmids. 
Accordingly, the amount of native KAR2 protein in cells expressing high 
levels of a gene product is diminished at least 5-fold. 
EXAMPLE 2 
Construction of an Inducible KAR2 Expression Vector 
A pMR1341 expression vector was made from a MR568 plasmid which encoded the 
yeast KAR2 chaperone protein having -55 base pairs (bp) from the ATG start 
codon (i.e. position 240 of SEQ ID NO: 1) to the terminus of the coding 
region at bp as provided in SEQ D NO:1. The PGAL1 promoter encoded within 
a SalI-AatII fragment from pB622 was placed into SalI-AatII sites within 
pMR568 to provide a galactose inducible promoter for the KAR2 coding 
region. Moreover, pMR1341 encodes a URA3 selectable marker which permits 
selection for this vector in ura deficient yeast host cells. In later 
experiments the URA3 encoding nucleic acid fragment was deleted and 
replaced with a fragment encoding both HIS and LEU yeast selectable 
markers. 
FIG. 2 depicts this pMR1341 expression vector for KAR2. As depicted, this 
vector encodes a pSC101 2origin of replication (ori pSC101) and an 
ampicillin resistance (Amp.sup.R) which permit replication and selection 
of pMR1341 in Escherichia coli. pMR1341 further encodes a yeast 
centromeric (CEN4) sequence and a yeast autonomous replication sequence-1 
(ARS1) which permit autonomous replication in yeast host cells. Vogel et 
al. (1990) describe this vector in greater detail. 
EXAMPLE 3 
Increased Secretion of Overexpressed Proteins Upon Expression of a 
Chaperone Protein 
The KAR2 yeast chaperone coding region was placed under the control of a 
galactose inducible promoter and the plasmid encoding this chimeric gene 
was transformed into BJ5464 yeast cells which also carried a plasmid 
encoding erythropoietin (EPO) under a galactose inducible promoter. These 
BJ5464 cells were then grown overnight in protein-free glucose medium in 
the absence of galactose. Expression of KAR2 and EPO proteins was induced 
by transfer of the BJ5464 cells into a galactose medium (SC GAL). 
Cell growth after induction was monitored by observing the optical 
absorption of the culture at 600 nm. Cell and supernatant samples were 
taken at 24, 48 and 72 hours after induction. Cell samples were used for 
determination of KAR2 protein levels using the slot blot procedure 
described in Example 1. Supernatant samples were tested for the amount of 
secreted EPO by using the slot blot procedure with a SY14 monoclonal 
antibody which is specific for EPO. 
FIG. 3 depicts the KAR2 expression observed in cell extracts collected at 
24, 48 and 72 hours after induction. The KAR2 immunoassay values provided 
in FIG. 3 represent a ratio of the amount of KAR2 detected in a given 
yeast cell type relative to wild type yeast. KAR2 expression in wild type 
cells (.box-solid.), cells transformed with the EPO-encoding plasmid only 
(.circle-solid., GalEpo) and cells transformed with both the EPO-encoding 
plasmid and the KAR2-encoding plasmid (.tangle-solidup., GalEpo+GalKar2), 
is depicted. After induction, expression of KAR2 is initially higher in 
cells with the EPO-encoding plasmid than in wild type yeast cells. 
However, GalEpo cellular expression of KAR2 drops to almost wild type 
levels by 48 hours after induction. If KAR2 expression were monitored for 
longer periods of time, the amount of KAR2 in the GalEPO cells would be 
less than wild type, as shown in FIG. 1. However, KAR2 expression at 24 hr 
is significantly greater in GalEpo+GalKAR2 cells which have the 
KAR2-encoding plasmid despite the presence of overexpressed EPO. Moreover, 
by 48 to 72 hours after induction, KAR2 expression is at least 4- to 
5-fold higher in cells expressing additional amounts of KAR2 recombinantly 
than in cells expressing KAR2 from a native, genomic locus. Therefore, 
KAR2 expression can be boosted significantly by recombinant expression. 
FIG. 4 depicts the growth of wild type cells (.quadrature.), cells 
transformed with the EPO-encoding plasmid only (.largecircle., GalEpo) and 
cells transformed with both the EPO-encoding plasmid and the KAR2-encoding 
plasmid (.DELTA., GalEpo+GalKar2) after induction of EPO and KAR2 
expression. 
The inset provided in FIG. 4 depicts the amount of EPO secreted into the 
medium of cells which have the EPO-encoding plasmid only (GalEpo) compared 
with the amount of secreted EPO from cells having both the EPO-encoding 
plasmid and the KAR2-encoding plasmid (GalEpo+GalKar2). The supernatants 
tested were collected during exponential growth of these yeast strains at 
the indicated time point (arrow). As shown in the FIG. 4 inset, the amount 
of EPO secreted upon induction of KAR2 expression is almost five-fold 
higher than when no additional KAR2 chaperone protein is present. 
Therefore, increasing KAR2 expression causes a substantial increase in 
protein secretion. 
EXAMPLE 4 
Construction of Strains Overexpressing BiP and PDI 
Yeast strains were constructed which overexpress yeast BiP, PDI or both BiP 
and PDI. 
The overexpression system for BiP utilizes the glyceraldehyde-3-phosphate 
dehydrogenase (GPD) constitutive promoter. A SalI-AatII fragment 
containing the GPD promoter was ligated into the AatII-SalI site of the 
pMRI341 expression vector described in Example 2, replacing the galactose 
(GAL1) promoter used for inducible expression of yeast BiP. A single-copy 
centromere plasmid containing this construct was named pGPDKAR2. BJ5464 
cells were transformed with pGPDKAR2. 
To construct a yeast strain that overexpresses yeast PDI, an expression 
cassette containing the yeast PDI gene downstream of the constitutive 
ADHII promoter was integrated into the chromosomal copy of PDI using LEU2 
as a selective marker. Yeast strain BJ5464 with this integrated PDI 
expression cassette was renamed YVH10 (PDI::ADHII-PDI-Leu2 ura3-52 trp 1 
leu2.DELTA.1 his 3.DELTA.200 pep4::H153 prb 1.DELTA.1.6p can 1 GAL). 
YVH10 cells were transformed with pGPDKAR2 to provide cells overexpressing 
both BiP and PDI. 
Cells extracts from mid-exponential phase cultures of BJ5464, BJ5464 
transformed with pGPDKAR2, YVH10, and YVH10 transformed with pGPDKAR2 were 
prepared. Yeast BiP and PDI were detected by chemiluminescence using 
.alpha.-Kar21gG and .alpha.-PDI1gG, respectively. Densitometry was 
performed with an Apple Optical Scanner and analyzed with the program 
Image (NIH). Quantitation of band intensity was determined from three 
dilutions of protein and multiple time exposures of the bands within the 
linear range of the film. 
As demonstrated in Table 3, BiP was overexpressed approximately 5-6 fold, 
and PDI was overexpressed approximately 11-16 fold. 
TABLE 3 
______________________________________ 
BJ5464 + YVH10 + 
BJ5464 pGFDKAR2 YVH10 GPDKAR2 
______________________________________ 
BiP - + - + 
overexpressed 
PDI - - + + 
overexpressed 
Densitometry 
1 5.9 1.3 5.5 
scan, .alpha.BiP 
Densitometry 
1.3 1 16 11 
scan, .alpha.PDI 
______________________________________ 
EXAMPLE 5 
Increased Secretion of Overexpressed Proteins Upon Expression of a 
Chaperone Protein 
The four yeast strains described in Example 4 (BJ5464, BJ5464+pGPDKAR2, 
YVH10, and YVH10+pGPDKAR2) are grown for several generations in synthetic 
complete (S.C.) media to provide strains which overexpress neither BiP nor 
PDI, BiP alone, PDI alone, or both BiP and PDI, respectively. The strains 
are each transformed with an expression vector which directs the 
constitutive expression of a gene product. Supernatant samples are 
collected during exponential growth of the transformed cells and assayed 
for the presence of the secreted gene product. 
__________________________________________________________________________ 
SEQUENCE LISTING 
(1) GENERAL INFORMATION: 
(iii) NUMBER OF SEQUENCES: 20 
(2) INFORMATION FOR SEQ ID NO:1: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 2780 base pairs 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: double 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(ix) FEATURE: 
(A) NAME/KEY: CDS 
(B) LOCATION: 285..2333 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:1: 
CTCGAGCAAAGTGTAGATCCCATTAGGACTCATCATTCATCTAATTTTGCTATGTTAGCT60 
GCAACTTTCTATTTTAATAGAACCTTCTGGAAATTTCACCCGGCGCGGCACCCGAGGAAC120 
TGGACAGCGTGTCGAAAAAGTTGCTTTTTTATATAAAGGACACGAAAAGGGTTCTCTGGA180 
AGATATAAATATGGCTATGTAATTCTAAAGATTAACGTGTTACTGTTTTACTTTTTTAAA240 
GTCCCCAAGAGTAGTCTCAAGGGAAAAAGCGTATCAAACATACCATGTTTTTCAAC296 
MetPhePheAsn 
AGACTAAGCGCTGGCAAGCTGCTGGTACCACTCTCCGTGGTCCTGTAC344 
ArgLeuSerAlaGlyLysLeuLeuValProLeuSerValValLeuTyr 
5101520 
GCCCTTTTCGTGGTAATATTACCTTTACAGAATTCTTTCCACTCCTCC392 
AlaLeuPheValValIleLeuProLeuGlnAsnSerPheHisSerSer 
253035 
AATGTTTTAGTTAGAGGTGCCGATGATGTAGAAAACTACGGAACTGTT440 
AsnValLeuValArgGlyAlaAspAspValGluAsnTyrGlyThrVal 
404550 
ATCGGTATTGACTTAGGTACTACTTATTCCTGTGTTGCTGTGATGAAA488 
IleGlyIleAspLeuGlyThrThrTyrSerCysValAlaValMetLys 
556065 
AATGGTAAGACTGAAATTCTTGCTAATGAGCAAGGTAACAGAATCACC536 
AsnGlyLysThrGluIleLeuAlaAsnGluGlnGlyAsnArgIleThr 
707580 
CCATCTTACGTGGCATTCACCGATGATGAAAGATTGATTGGTGATGCT584 
ProSerTyrValAlaPheThrAspAspGluArgLeuIleGlyAspAla 
859095100 
GCAAAGAACCAAGTTGCTGCCAATCCTCAAAACACCATCTTCGACATT632 
AlaLysAsnGlnValAlaAlaAsnProGlnAsnThrIlePheAspIle 
105110115 
AAGAGATTGATCGGTTTGAAATATAACGACAGATCTGTTCAGAAGGAT680 
LysArgLeuIleGlyLeuLysTyrAsnAspArgSerValGlnLysAsp 
120125130 
ATCAAGCACTTGCCATTTAATGTGGTTAATAAAGATGGGAAGCCCGCT728 
IleLysHisLeuProPheAsnValValAsnLysAspGlyLysProAla 
135140145 
GTAGAAGTAAGTGTCAAAGGAGAAAAGAAGGTTTTTACTCCAGAAGAA776 
ValGluValSerValLysGlyGluLysLysValPheThrProGluGlu 
150155160 
ATTTCTGGTATGATCTTGGGTAAGATGAAACAAATTGCCGAAGATTAT824 
IleSerGlyMetIleLeuGlyLysMetLysGlnIleAlaGluAspTyr 
165170175180 
TTAGGCACTAAGGTTACCCATGCTGTCGTTACTGTTCCTGCTTATTTC872 
LeuGlyThrLysValThrHisAlaValValThrValProAlaTyrPhe 
185190195 
AATGACGCGCAAAGACAAGCCACCAAGGATGCTGGTACCATCGCTGGT920 
AsnAspAlaGlnArgGlnAlaThrLysAspAlaGlyThrIleAlaGly 
200205210 
TTGAACGTTTTGAGAATTGTTAATGAACCAACCGCAGCCGCCATTGCC968 
LeuAsnValLeuArgIleValAsnGluProThrAlaAlaAlaIleAla 
215220225 
TACGGTTTGGATAAATCTGATAAGGAACATCAAATTATTGTTTATGAT1016 
TyrGlyLeuAspLysSerAspLysGluHisGlnIleIleValTyrAsp 
230235240 
TTGGGTGGTGGTACTTTCGATGTCTCTCTATTGTCTATTGAAAACGGT1064 
LeuGlyGlyGlyThrPheAspValSerLeuLeuSerIleGluAsnGly 
245250255260 
GTTTTCGAAGTCCAAGCCACTTCTGGTGATACTCATTTAGGTGGTGAA1112 
ValPheGluValGlnAlaThrSerGlyAspThrHisLeuGlyGlyGlu 
265270275 
GATTTTGACTATAAGATCGTTCGTCAATTGATAAAAGCTTTCAAGAAG1160 
AspPheAspTyrLysIleValArgGlnLeuIleLysAlaPheLysLys 
280285290 
AAGCATGGTATTGATGTGTCTGACAACAACAAGGCCCTAGCTAAATTG1208 
LysHisGlyIleAspValSerAspAsnAsnLysAlaLeuAlaLysLeu 
295300305 
AAGAGAGAAGCTGAAAAGGCTAAACGTGCCTTGTCCAGCCAAATGTCC1256 
LysArgGluAlaGluLysAlaLysArgAlaLeuSerSerGlnMetSer 
310315320 
ACCCGTATTGAAATTGACTCCTTCGTTGATGGTATCGACTTAAGTGAA1304 
ThrArgIleGluIleAspSerPheValAspGlyIleAspLeuSerGlu 
325330335340 
ACCTTGACCAGAGCTAAGTTTGAGGAATTAAACCTAGATCTATTCAAG1352 
ThrLeuThrArgAlaLysPheGluGluLeuAsnLeuAspLeuPheLys 
345350355 
AAGACCTTGAAGCCTGTCGAGAAGGTTTTGCAAGATTCTGGTTTGGAA1400 
LysThrLeuLysProValGluLysValLeuGlnAspSerGlyLeuGlu 
360365370 
AAGAAGGATGTTGATGATATCGTTTTGGTTGGTGGTTCTACTAGAATT1448 
LysLysAspValAspAspIleValLeuValGlyGlySerThrArgIle 
375380385 
CCAAAGGTCCAACAATTGTTAGAATCATACTTTGATGGTAAGAAGGCC1496 
ProLysValGlnGlnLeuLeuGluSerTyrPheAspGlyLysLysAla 
390395400 
TCCAAGGGTATTAACCCAGATGAAGCTGTTGCATACGGTGCAGCCGTT1544 
SerLysGlyIleAsnProAspGluAlaValAlaTyrGlyAlaAlaVal 
405410415420 
CAAGCTGGTGTCTTATCCGGTGAAGAAGGTGTCGAAGATATTGTTTTA1592 
GlnAlaGlyValLeuSerGlyGluGluGlyValGluAspIleValLeu 
425430435 
TTGGATGTCAACGCTTTGACTCTTGGTATTGAAACCACTGGTGGTGTC1640 
LeuAspValAsnAlaLeuThrLeuGlyIleGluThrThrGlyGlyVal 
440445450 
ATGACTCCATTAATTAAGAGAAATACTGCTATTCCTACAAAGAAATCC1688 
MetThrProLeuIleLysArgAsnThrAlaIleProThrLysLysSer 
455460465 
CAAATTTTCTCTACTGCCGTTGACAACCAACCAACCGTTATGATCAAG1736 
GlnIlePheSerThrAlaValAspAsnGlnProThrValMetIleLys 
470475480 
GTATACGAGGGTGAAAGAGCCATGTCTAAGGACAACAATCTATTAGGT1784 
ValTyrGluGlyGluArgAlaMetSerLysAspAsnAsnLeuLeuGly 
485490495500 
AAGTTTGAATTAACCGGCATTCCACCAGCACCAAGAGGTGTACCTCAA1832 
LysPheGluLeuThrGlyIleProProAlaProArgGlyValProGln 
505510515 
ATTGAAGTCACATTTGCACTTGACGCTAATGGTATTCTGAAGGTGTCT1880 
IleGluValThrPheAlaLeuAspAlaAsnGlyIleLeuLysValSer 
520525530 
GCCACAGATAAGGGAACTGGTAAATCCGAATCTATCACCATCACTAAC1928 
AlaThrAspLysGlyThrGlyLysSerGluSerIleThrIleThrAsn 
535540545 
GATAAAGGTAGATTAACCCAAGAAGAGATTGATAGAATGGTTGAAGAG1976 
AspLysGlyArgLeuThrGlnGluGluIleAspArgMetValGluGlu 
550555560 
GCTGAAAAATTCGCTTCTGAAGACGCTTCTATCAAGGCCAAGGTTGAA2024 
AlaGluLysPheAlaSerGluAspAlaSerIleLysAlaLysValGlu 
565570575580 
TCTAGAAACAAATTAGAAAACTACGCTCACTCTTTGAAAAACCAAGTT2072 
SerArgAsnLysLeuGluAsnTyrAlaHisSerLeuLysAsnGlnVal 
585590595 
AATGGTGACCTAGGTGAAAAATTGGAAGAAGAAGACAAGGAAACCTTA2120 
AsnGlyAspLeuGlyGluLysLeuGluGluGluAspLysGluThrLeu 
600605610 
TTAGATGCTGCTAACGATGTTTTAGAATGGTTAGATGATAACTTTGAA2168 
LeuAspAlaAlaAsnAspValLeuGluTrpLeuAspAspAsnPheGlu 
615620625 
ACCGCCATTGCTGAAGACTTTGATGAAAAGTTCGAATCTTTGTCCAAG2216 
ThrAlaIleAlaGluAspPheAspGluLysPheGluSerLeuSerLys 
630635640 
GTCGCTTATCCAATTACTTCTAAGTTGTACGGAGGTGCTGATGGTTCT2264 
ValAlaTyrProIleThrSerLysLeuTyrGlyGlyAlaAspGlySer 
645650655660 
GGTGCCGCTGATTATGACGACGAAGATGAAGATGACGATGGTGATTAT2312 
GlyAlaAlaAspTyrAspAspGluAspGluAspAspAspGlyAspTyr 
665670675 
TTCGAACACGACGAATTGTAGATAAAATAGTTAAAAATTTTTGCTGCT2360 
PheGluHisAspGluLeu 
680 
GGAAGCTTCAAGGTTGTTAATTTATTGACTTGCATAGAATATCTACATTTCTTCTAAAAA2420 
TACATGCATAGCTAATTCAAACTTCGAGCTTCATACAATTTTCGAGGAGATTATACTGAG2480 
TATATACGTAAATATATGCATTATATGTTATAAAATTAGAAAGATATAGAAATTTCATTG2540 
AAGAGTATAGAGACTGGGGTTAAGGTACTCAGTAACAGTGTCATCAATATGCTAATTTTG2600 
CGTATTACTTAGCTCTATTGCGCAAATGCAATTTTTTCTTACCCTGATAATGCTTTATTT2660 
CCCGTTCCGAAAATTTTTCACTGAAAAAAAAGTGCTTAAGCTCATCTCATCTCATCTCAT2720 
CCCATCACTATTGAAATATTTTGCTAAAACATTATAACAGAGAGAGTTGAAAGGCTCGAG2780 
(2) INFORMATION FOR SEQ ID NO:2: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 682 amino acids 
(B) TYPE: amino acid 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: 
MetPhePheAsnArgLeuSerAlaGlyLysLeuLeuValProLeuSer 
151015 
ValValLeuTyrAlaLeuPheValValIleLeuProLeuGlnAsnSer 
202530 
PheHisSerSerAsnValLeuValArgGlyAlaAspAspValGluAsn 
354045 
TyrGlyThrValIleGlyIleAspLeuGlyThrThrTyrSerCysVal 
505560 
AlaValMetLysAsnGlyLysThrGluIleLeuAlaAsnGluGlnGly 
65707580 
AsnArgIleThrProSerTyrValAlaPheThrAspAspGluArgLeu 
859095 
IleGlyAspAlaAlaLysAsnGlnValAlaAlaAsnProGlnAsnThr 
100105110 
IlePheAspIleLysArgLeuIleGlyLeuLysTyrAsnAspArgSer 
115120125 
ValGlnLysAspIleLysHisLeuProPheAsnValValAsnLysAsp 
130135140 
GlyLysProAlaValGluValSerValLysGlyGluLysLysValPhe 
145150155160 
ThrProGluGluIleSerGlyMetIleLeuGlyLysMetLysGlnIle 
165170175 
AlaGluAspTyrLeuGlyThrLysValThrHisAlaValValThrVal 
180185190 
ProAlaTyrPheAsnAspAlaGlnArgGlnAlaThrLysAspAlaGly 
195200205 
ThrIleAlaGlyLeuAsnValLeuArgIleValAsnGluProThrAla 
210215220 
AlaAlaIleAlaTyrGlyLeuAspLysSerAspLysGluHisGlnIle 
225230235240 
IleValTyrAspLeuGlyGlyGlyThrPheAspValSerLeuLeuSer 
245250255 
IleGluAsnGlyValPheGluValGlnAlaThrSerGlyAspThrHis 
260265270 
LeuGlyGlyGluAspPheAspTyrLysIleValArgGlnLeuIleLys 
275280285 
AlaPheLysLysLysHisGlyIleAspValSerAspAsnAsnLysAla 
290295300 
LeuAlaLysLeuLysArgGluAlaGluLysAlaLysArgAlaLeuSer 
305310315320 
SerGlnMetSerThrArgIleGluIleAspSerPheValAspGlyIle 
325330335 
AspLeuSerGluThrLeuThrArgAlaLysPheGluGluLeuAsnLeu 
340345350 
AspLeuPheLysLysThrLeuLysProValGluLysValLeuGlnAsp 
355360365 
SerGlyLeuGluLysLysAspValAspAspIleValLeuValGlyGly 
370375380 
SerThrArgIleProLysValGlnGlnLeuLeuGluSerTyrPheAsp 
385390395400 
GlyLysLysAlaSerLysGlyIleAsnProAspGluAlaValAlaTyr 
405410415 
GlyAlaAlaValGlnAlaGlyValLeuSerGlyGluGluGlyValGlu 
420425430 
AspIleValLeuLeuAspValAsnAlaLeuThrLeuGlyIleGluThr 
435440445 
ThrGlyGlyValMetThrProLeuIleLysArgAsnThrAlaIlePro 
450455460 
ThrLysLysSerGlnIlePheSerThrAlaValAspAsnGlnProThr 
465470475480 
ValMetIleLysValTyrGluGlyGluArgAlaMetSerLysAspAsn 
485490495 
AsnLeuLeuGlyLysPheGluLeuThrGlyIleProProAlaProArg 
500505510 
GlyValProGlnIleGluValThrPheAlaLeuAspAlaAsnGlyIle 
515520525 
LeuLysValSerAlaThrAspLysGlyThrGlyLysSerGluSerIle 
530535540 
ThrIleThrAsnAspLysGlyArgLeuThrGlnGluGluIleAspArg 
545550555560 
MetValGluGluAlaGluLysPheAlaSerGluAspAlaSerIleLys 
565570575 
AlaLysValGluSerArgAsnLysLeuGluAsnTyrAlaHisSerLeu 
580585590 
LysAsnGlnValAsnGlyAspLeuGlyGluLysLeuGluGluGluAsp 
595600605 
LysGluThrLeuLeuAspAlaAlaAsnAspValLeuGluTrpLeuAsp 
610615620 
AspAsnPheGluThrAlaIleAlaGluAspPheAspGluLysPheGlu 
625630635640 
SerLeuSerLysValAlaTyrProIleThrSerLysLeuTyrGlyGly 
645650655 
AlaAspGlySerGlyAlaAlaAspTyrAspAspGluAspGluAspAsp 
660665670 
AspGlyAspTyrPheGluHisAspGluLeu 
675680 
(2) INFORMATION FOR SEQ ID NO:3: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 2367 base pairs 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: double 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(ix) FEATURE: 
(A) NAME/KEY: CDS 
(B) LOCATION: 251..2176 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: 
AAGCTTTTAGGAATTTTGAATTTTTGATCGAATTTTAGAAAAAACTATTCGCAAGACTAC60 
AATTTTTGAAGGGTGCTATTTGTGAAAAAATAAAACGTGAAATAAATCGTTTTATAATTT120 
ACGAATTGTCGTTATTCAAAACTCAAAAAATATGATCTCGTCGAGATTCACTAATGTAGT180 
CCGTAGCGGATTGCGTTTCCAAAGCAAGGGAGCATCGTTCAAGATTGGCGCTTCCTTGCA240 
TGGAAGTCGCATGACCGCCCGCTGGAATTCTAATGCAAGTGGTAATGAA289 
MetThrAlaArgTrpAsnSerAsnAlaSerGlyAsnGlu 
1510 
AAAGTTAAGGGTCCCGTAATCGGTATTGACTTGGGTACCACCACCTCA337 
LysValLysGlyProValIleGlyIleAspLeuGlyThrThrThrSer 
152025 
TGTTTAGCAATCATGGAGGGTCAAACCCCTAAGGTTATTGCAAATGCC385 
CysLeuAlaIleMetGluGlyGlnThrProLysValIleAlaAsnAla 
30354045 
GAGGGTACCCGTACCACACCATCTGTCGTCGCATTTACCAAAGATGGC433 
GluGlyThrArgThrThrProSerValValAlaPheThrLysAspGly 
505560 
GAGCGTTTGGTGGGTGTTAGCGCTAAACGCCAAGCCGTCATTAACCCG481 
GluArgLeuValGlyValSerAlaLysArgGlnAlaValIleAsnPro 
657075 
GAAAACACATTTTTTGCTACTAAGCGTTTAATCGGTCGTAGATTTAAA529 
GluAsnThrPhePheAlaThrLysArgLeuIleGlyArgArgPheLys 
808590 
GAGCCTGAAGTCCAACGTGATATTAAGGAAGTTCCTTACAAAATTGTC577 
GluProGluValGlnArgAspIleLysGluValProTyrLysIleVal 
95100105 
GAGCACTCAAATGGAGATGCTTGGTTGGAGGCTCGTGGTAAGACCTAC625 
GluHisSerAsnGlyAspAlaTrpLeuGluAlaArgGlyLysThrTyr 
110115120125 
TCTCCATCTCAAATCGGTGGTTTCATCCTTAGTAAGATGAGGGAAACT673 
SerProSerGlnIleGlyGlyPheIleLeuSerLysMetArgGluThr 
130135140 
GCCAGCACCTACCTTGGAAAAGATGTAAAGAATGCCGTTGTTACTGTT721 
AlaSerThrTyrLeuGlyLysAspValLysAsnAlaValValThrVal 
145150155 
CCTGCTTACTTCAATGACTCTCAGCGTCAAGCTACCAAGGCTGCTGGT769 
ProAlaTyrPheAsnAspSerGlnArgGlnAlaThrLysAlaAlaGly 
160165170 
GCCATTGCTGGTTTGAATGTTTTGCGTGTCGTCAACGAGCCTACTGCC817 
AlaIleAlaGlyLeuAsnValLeuArgValValAsnGluProThrAla 
175180185 
GCCGCTTTGGCTTATGGTTTGGACAAGAAGAATGATGCCATCGTCGCA865 
AlaAlaLeuAlaTyrGlyLeuAspLysLysAsnAspAlaIleValAla 
190195200205 
GTTTTCGATTTGGGTGGTGGTACTTTTGATATTTCTATTTTGGAGTTA913 
ValPheAspLeuGlyGlyGlyThrPheAspIleSerIleLeuGluLeu 
210215220 
AACAATGGTGTTTTTGAGGTTAGAAGTACCAACGGTGACACTCATTTG961 
AsnAsnGlyValPheGluValArgSerThrAsnGlyAspThrHisLeu 
225230235 
GGTGGTGAGGACTTTGATGTTGCTCTTGTTCGTCACATTGTCGAGACC1009 
GlyGlyGluAspPheAspValAlaLeuValArgHisIleValGluThr 
240245250 
TTTAAGAAGAATGAGGGTTTGGACTTGAGCAAGGACCGTCTCGCCGTT1057 
PheLysLysAsnGluGlyLeuAspLeuSerLysAspArgLeuAlaVal 
255260265 
CAACGTATTCGTGAGGCTGCTGAAAAAGCTAAGTGCGAACTTTCCTCT1105 
GlnArgIleArgGluAlaAlaGluLysAlaLysCysGluLeuSerSer 
270275280285 
CTTTCCAAGACTGATATCAGTCTTCCTTTCATTACTGCGGATGCTACT1153 
LeuSerLysThrAspIleSerLeuProPheIleThrAlaAspAlaThr 
290295300 
GGCCCTAAGCATATTAACATGGAAATCTCTCGTGCTCAATTTGAGAAA1201 
GlyProLysHisIleAsnMetGluIleSerArgAlaGlnPheGluLys 
305310315 
CTTGTTGATCCTCTCGTTCGTCGTACCATCGATCCTTGCAAGCGTGCC1249 
LeuValAspProLeuValArgArgThrIleAspProCysLysArgAla 
320325330 
CTTAAGGATGCTAACTTGCAAACCTCTGAAATCAATGAAGTTATCCTT1297 
LeuLysAspAlaAsnLeuGlnThrSerGluIleAsnGluValIleLeu 
335340345 
GTCGGTGGTATGACTCGTATGCCTCGTGTTGTCGAAACTGTCAAGAGT1345 
ValGlyGlyMetThrArgMetProArgValValGluThrValLysSer 
350355360365 
ATCTTCAAGCGTGAACCCGCTAAGTCCGTCAACCCTGATGAAGCTGTT1393 
IlePheLysArgGluProAlaLysSerValAsnProAspGluAlaVal 
370375380 
GCCATTGGTGCTGCTATTCAAGGTGGTGTCTTGTCTGGCCATGTTAAG1441 
AlaIleGlyAlaAlaIleGlnGlyGlyValLeuSerGlyHisValLys 
385390395 
GACCTTGTTCTTTTGGATGTCACCCCCTTGTCCCTCGGTATCGAGACT1489 
AspLeuValLeuLeuAspValThrProLeuSerLeuGlyIleGluThr 
400405410 
TTGGGCGGTGTTTTCACTCGTTTGATCAACCGTAACACTACCATTCCT1537 
LeuGlyGlyValPheThrArgLeuIleAsnArgAsnThrThrIlePro 
415420425 
ACTCGCAAGTCTCAAGTTTTCTCCACTGCTGCTGATGGTCAAACTGCC1585 
ThrArgLysSerGlnValPheSerThrAlaAlaAspGlyGlnThrAla 
430435440445 
GTTGAAATCCGTGTCTTCCAGGGTGAACGTGAGCTTGTTCGTGACAAC1633 
ValGluIleArgValPheGlnGlyGluArgGluLeuValArgAspAsn 
450455460 
AAATTAATTGGCAACTTCCAACTTACTGGCATTGCTCCTGCACCTAAG1681 
LysLeuIleGlyAsnPheGlnLeuThrGlyIleAlaProAlaProLys 
465470475 
GGTCAACCTCAGATTGAGGTTTCTTTTGATGTTGATGCCGATGGCATT1729 
GlyGlnProGlnIleGluValSerPheAspValAspAlaAspGlyIle 
480485490 
ATCAATGTCTCTGCCCGTGACAAGGCTACCAACAAGGATTCTTCCATC1777 
IleAsnValSerAlaArgAspLysAlaThrAsnLysAspSerSerIle 
495500505 
ACTGTTGCTGGATCTTCCGGTTTAACTGATTCTGAGATTGAGGCTATG1825 
ThrValAlaGlySerSerGlyLeuThrAspSerGluIleGluAlaMet 
510515520525 
GTTGCCGATGCTGAGAAGTATCGTGCCAGTGACATGGCTCGCAAGGAG1873 
ValAlaAspAlaGluLysTyrArgAlaSerAspMetAlaArgLysGlu 
530535540 
GCTATTGAGAACGGAAACAGAGCTGAAAGCGTCTGCACCGATATTGAA1921 
AlaIleGluAsnGlyAsnArgAlaGluSerValCysThrAspIleGlu 
545550555 
AGCAACCTTGACATTCACAAAGACAAATTGGACCAACAAGCTGTTGAA1969 
SerAsnLeuAspIleHisLysAspLysLeuAspGlnGlnAlaValGlu 
560565570 
GACTTGCGCTCCAAGATCACCGATCTCCGTGAAACTGTTGCCAAGGTC2017 
AspLeuArgSerLysIleThrAspLeuArgGluThrValAlaLysVal 
575580585 
AACGCTGGTGACGAAGGTATTACTAGTGAAGATATGAAGAAGAAGATT2065 
AsnAlaGlyAspGluGlyIleThrSerGluAspMetLysLysLysIle 
590595600605 
GATGAAATTCAACAACTCTCTTTGAAGGTTTTCGAGTCTGTCTACAAG2113 
AspGluIleGlnGlnLeuSerLeuLysValPheGluSerValTyrLys 
610615620 
AACCAAAATCAAGGTAATGAATCTTCTGGTGATAACTCTGCTCCTGAG2161 
AsnGlnAsnGlnGlyAsnGluSerSerGlyAspAsnSerAlaProGlu 
625630635 
GGTGACAAGAAGTAGAGTGCACACCACAGTACGAAATGACATGTGCAATTTT2213 
GlyAspLysLys 
640 
CAATTTTAGCTCTATATGTCAAAAAATTTATGTGGATAATTGATTATCCATTTACATGTT2273 
GAAAGAAAATGTCTGGATTTTGAAAAGGTAAACTATGATATTTTTATTAAATGTTCTAAA2333 
AAAAAAAAAAAAAAAAAAAAAAAAACCGGAATTC2367 
(2) INFORMATION FOR SEQ ID NO:4: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 641 amino acids 
(B) TYPE: amino acid 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: 
MetThrAlaArgTrpAsnSerAsnAlaSerGlyAsnGluLysValLys 
151015 
GlyProValIleGlyIleAspLeuGlyThrThrThrSerCysLeuAla 
202530 
IleMetGluGlyGlnThrProLysValIleAlaAsnAlaGluGlyThr 
354045 
ArgThrThrProSerValValAlaPheThrLysAspGlyGluArgLeu 
505560 
ValGlyValSerAlaLysArgGlnAlaValIleAsnProGluAsnThr 
65707580 
PhePheAlaThrLysArgLeuIleGlyArgArgPheLysGluProGlu 
859095 
ValGlnArgAspIleLysGluValProTyrLysIleValGluHisSer 
100105110 
AsnGlyAspAlaTrpLeuGluAlaArgGlyLysThrTyrSerProSer 
115120125 
GlnIleGlyGlyPheIleLeuSerLysMetArgGluThrAlaSerThr 
130135140 
TyrLeuGlyLysAspValLysAsnAlaValValThrValProAlaTyr 
145150155160 
PheAsnAspSerGlnArgGlnAlaThrLysAlaAlaGlyAlaIleAla 
165170175 
GlyLeuAsnValLeuArgValValAsnGluProThrAlaAlaAlaLeu 
180185190 
AlaTyrGlyLeuAspLysLysAsnAspAlaIleValAlaValPheAsp 
195200205 
LeuGlyGlyGlyThrPheAspIleSerIleLeuGluLeuAsnAsnGly 
210215220 
ValPheGluValArgSerThrAsnGlyAspThrHisLeuGlyGlyGlu 
225230235240 
AspPheAspValAlaLeuValArgHisIleValGluThrPheLysLys 
245250255 
AsnGluGlyLeuAspLeuSerLysAspArgLeuAlaValGlnArgIle 
260265270 
ArgGluAlaAlaGluLysAlaLysCysGluLeuSerSerLeuSerLys 
275280285 
ThrAspIleSerLeuProPheIleThrAlaAspAlaThrGlyProLys 
290295300 
HisIleAsnMetGluIleSerArgAlaGlnPheGluLysLeuValAsp 
305310315320 
ProLeuValArgArgThrIleAspProCysLysArgAlaLeuLysAsp 
325330335 
AlaAsnLeuGlnThrSerGluIleAsnGluValIleLeuValGlyGly 
340345350 
MetThrArgMetProArgValValGluThrValLysSerIlePheLys 
355360365 
ArgGluProAlaLysSerValAsnProAspGluAlaValAlaIleGly 
370375380 
AlaAlaIleGlnGlyGlyValLeuSerGlyHisValLysAspLeuVal 
385390395400 
LeuLeuAspValThrProLeuSerLeuGlyIleGluThrLeuGlyGly 
405410415 
ValPheThrArgLeuIleAsnArgAsnThrThrIleProThrArgLys 
420425430 
SerGlnValPheSerThrAlaAlaAspGlyGlnThrAlaValGluIle 
435440445 
ArgValPheGlnGlyGluArgGluLeuValArgAspAsnLysLeuIle 
450455460 
GlyAsnPheGlnLeuThrGlyIleAlaProAlaProLysGlyGlnPro 
465470475480 
GlnIleGluValSerPheAspValAspAlaAspGlyIleIleAsnVal 
485490495 
SerAlaArgAspLysAlaThrAsnLysAspSerSerIleThrValAla 
500505510 
GlySerSerGlyLeuThrAspSerGluIleGluAlaMetValAlaAsp 
515520525 
AlaGluLysTyrArgAlaSerAspMetAlaArgLysGluAlaIleGlu 
530535540 
AsnGlyAsnArgAlaGluSerValCysThrAspIleGluSerAsnLeu 
545550555560 
AspIleHisLysAspLysLeuAspGlnGlnAlaValGluAspLeuArg 
565570575 
SerLysIleThrAspLeuArgGluThrValAlaLysValAsnAlaGly 
580585590 
AspGluGlyIleThrSerGluAspMetLysLysLysIleAspGluIle 
595600605 
GlnGlnLeuSerLeuLysValPheGluSerValTyrLysAsnGlnAsn 
610615620 
GlnGlyAsnGluSerSerGlyAspAsnSerAlaProGluGlyAspLys 
625630635640 
Lys 
(2) INFORMATION FOR SEQ ID NO:5: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 679 amino acids 
(B) TYPE: amino acid 
(C) STRANDEDNESS: single 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: 
MetPheSerAlaArgLysSerSerValGlyTrpLeuValSerSerLeu 
151015 
AlaValPheTyrValLeuLeuAlaValIleMetProIleAlaLeuThr 
202530 
GlySerGlnSerSerArgValValAlaArgAlaAlaGluAspHisGlu 
354045 
AspTyrGlyThrValIleGlyIleAspLeuGlyThrThrTyrSerCys 
505560 
ValAlaValMetLysAsnGlyLysThrGluIleLeuAlaAsnGluGln 
65707580 
GlyAsnArgIleThrProSerTyrValSerPheThrAspAspGluArg 
859095 
LeuIleGlyAspAlaAlaLysAsnGlnAlaAlaSerAsnProLysAsn 
100105110 
ThrIlePheAspIleLysArgLeuIleGlyLeuGlnTyrAsnAspPro 
115120125 
ThrValGlnArgAspIleLysHisLeuProTyrThrValValAsnLys 
130135140 
GlyAsnLysProTyrValGluValThrValLysGlyGluLysLysGlu 
145150155160 
PheThrProGluGluValSerGlyMetIleLeuGlyLysMetLysGln 
165170175 
IleAlaGluAspTyrLeuGlyLysLysValThrHisAlaValValThr 
180185190 
ValProAlaTyrPheAsnAspAlaGlnArgGlnAlaThrLysAspAla 
195200205 
GlyAlaIleAlaGlyLeuAsnIleLeuArgIleValAsnGluProThr 
210215220 
AlaAlaAlaIleAlaTyrGlyLeuAspLysThrGluAspGluHisGln 
225230235240 
IleIleValTyrAspLeuGlyGlyGlyThrPheAspValSerLeuLeu 
245250255 
SerIleGluAsnGlyValPheGluValGlnAlaThrAlaGlyAspThr 
260265270 
HisLeuGlyGlyGluAspPheAspTyrLysLeuValArgHisPheAla 
275280285 
GlnLeuPheGlnLysLysHisAspLeuAspValThrLysAsnAspLys 
290295300 
AlaMetAlaLysLeuLysArgGluAlaGluLysAlaLysArgSerLeu 
305310315320 
SerSerGlnThrSerThrArgIleGluIleAspSerPhePheAsnGly 
325330335 
IleAspPheSerGluThrLeuThrArgAlaLysPheGluGluLeuAsn 
340345350 
LeuAlaLeuPheLysLysThrLeuLysProValGluLysValLeuLys 
355360365 
AspSerGlyLeuGlnLysGluAspIleAspAspIleValLeuValGly 
370375380 
GlySerThrArgIleProLysValGlnGlnLeuLeuGluLysPhePhe 
385390395400 
AsnGlyLysLysAlaSerLysGlyIleAsnProAspGluAlaValAla 
405410415 
TyrGlyAlaAlaValGlnAlaGlyValLeuSerGlyGluGluGlyVal 
420425430 
GluAspIleValLeuLeuAspValAsnAlaLeuThrLeuGlyIleGlu 
435440445 
ThrThrGlyGlyValMetThrProLeuIleLysArgAsnThrAlaIle 
450455460 
ProThrLysLysSerGlnIlePheSerThrAlaValAspAsnGlnLys 
465470475480 
AlaValArgIleGlnValTyrGluGlyGluArgAlaMetValLysAsp 
485490495 
AsnAsnLeuLeuGlyAsnPheGluLeuSerAspIleArgAlaAlaPro 
500505510 
ArgGlyValProGlnIleGluValThrPheAlaLeuAspAlaAsnGly 
515520525 
IleLeuThrValSerAlaThrAspLysAspThrGlyLysSerGluSer 
530535540 
IleThrIleAlaAsnAspLysGlyArgLeuSerGlnAspAspIleAsp 
545550555560 
ArgMetValGluGluAlaGluLysTyrAlaAlaGluAspAlaLysPhe 
565570575 
LysAlaLysSerGluAlaArgAsnThrPheGluAsnPheValHisTyr 
580585590 
ValLysAsnSerValAsnGlyGluLeuAlaGluIleMetAspGluAsp 
595600605 
AspLysGluThrValLeuAspAsnValAsnGluSerLeuGluTrpLeu 
610615620 
GluAspAsnSerAspValAlaGluAlaGluAspPheGluGluLysMet 
625630635640 
AlaSerPheLysGluSerValGluProIleLeuAlaLysAlaSerAla 
645650655 
SerGlnGlySerThrSerGlyGluGlyPheGluAspGluAspAspAsp 
660665670 
AspTyrPheAspAspGluLeu 
675 
(2) INFORMATION FOR SEQ ID NO:6: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 2574 base pairs 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: double 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(ix) FEATURE: 
(A) NAME/KEY: CDS 
(B) LOCATION: 441..2429 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: 
CACAATATCAATAAGTTCCACTCACGCTTTGTCTTTCACAATATCATTTCAGAATTTACC60 
AATTTCGATTTTCATTGTTACATTCATTGCTATGAAAACGTAAGGTGGTGGCGGCAATAG120 
GACTTATCGAAATGTACAGAACTCACTATAGAATTGTTGTGTTGATGAGCTTCAACTGCA180 
TTCTTCTGGAAAGTACTAGTATTAACGACGTGACTGCTCCTCTCGTTACTTAGCTGATTT240 
CTGGTACGCTATTAAACTCATCCAAAACCAACTATTCTAGTTTGGTAAATCTTAATCAAA300 
AACTATTAAAACCCGTTTACTATTTACTTAACAGGTTGTTTTCAATAATTGGGAATTGCT360 
TGTGCCTACGATCTCTTGTAATTGAACTACACATATAAGCATTTATAAGTTGGTAATCTT420 
CAAATTCTTGTTTATTGAAAATGAAGAAGTTCCAGCTATTTAGCATTTTA470 
MetLysLysPheGlnLeuPheSerIleLeu 
1510 
AGCTACTTTGTAGCTTTATTCCTCCTACCTATGGCTTTTGCTAGTGGT518 
SerTyrPheValAlaLeuPheLeuLeuProMetAlaPheAlaSerGly 
152025 
GATGATAACTCTACAGAATCATATGGAACAGTTATTGGTATTGATCTT566 
AspAspAsnSerThrGluSerTyrGlyThrValIleGlyIleAspLeu 
303540 
GGTACAACATACTCTTGCGTTGCCGTTATGAAAAATGGTCGTGTAGAA614 
GlyThrThrTyrSerCysValAlaValMetLysAsnGlyArgValGlu 
455055 
ATTATTGCCAACGATCAGGGTAATCGTATTACACCCTCATATGTGGCC662 
IleIleAlaAsnAspGlnGlyAsnArgIleThrProSerTyrValAla 
606570 
TTTACTGAAGACGAACGTTTGGTTGGTGAGGCCGCTAAGAACCAAGCT710 
PheThrGluAspGluArgLeuValGlyGluAlaAlaLysAsnGlnAla 
75808590 
CCTTCCAATCCTGAAAACACCATTTTTGACATCAAGCGTCTTATTGGA758 
ProSerAsnProGluAsnThrIlePheAspIleLysArgLeuIleGly 
95100105 
CGTAAGTTTGACGAAAAGACAATGGCCAAGGATATTAAATCTTTTCCT806 
ArgLysPheAspGluLysThrMetAlaLysAspIleLysSerPhePro 
110115120 
TTCCATATTGTAAATGACAAGAACCGTCCTTTGGTTGAGGTTAATGTA854 
PheHisIleValAsnAspLysAsnArgProLeuValGluValAsnVal 
125130135 
GGTGGTAAGAAGAAAAAGTTTACCCCTGAAGAAATTTCAGCCATGATT902 
GlyGlyLysLysLysLysPheThrProGluGluIleSerAlaMetIle 
140145150 
CTTAGTAAAATGAAGCAAACTGCTGAAGCTTACCTCGGAAAGCCTGTC950 
LeuSerLysMetLysGlnThrAlaGluAlaTyrLeuGlyLysProVal 
155160165170 
ACTCACTCTGTTGTTACTGTCCCCGCCTACTTCAATGACGCTCAGCGT998 
ThrHisSerValValThrValProAlaTyrPheAsnAspAlaGlnArg 
175180185 
CAGGCTACCAAGGATGCTGGTACTATTGCCGGCTTGAATGTTATTCGT1046 
GlnAlaThrLysAspAlaGlyThrIleAlaGlyLeuAsnValIleArg 
190195200 
ATCGTCAATGAGCCTACTGCGGCTGCTATTGCCTACGGATTAGACAAA1094 
IleValAsnGluProThrAlaAlaAlaIleAlaTyrGlyLeuAspLys 
205210215 
ACTGATACAGAGAAGCATATTGTTGTTTATGATTTAGGTGGTGGTACT1142 
ThrAspThrGluLysHisIleValValTyrAspLeuGlyGlyGlyThr 
220225230 
TTTGACGTTTCTCTTTTGTCTATTGACAATGGTGTTTTCGAAGTTTTG1190 
PheAspValSerLeuLeuSerIleAspAsnGlyValPheGluValLeu 
235240245250 
GCTACTTCAGGTGATACCCATCTCGGTGGTGAGGACTTTGACAACCGT1238 
AlaThrSerGlyAspThrHisLeuGlyGlyGluAspPheAspAsnArg 
255260265 
GTTATCAACTACTTAGCCCGTACTTACAACCGCAAGAACAATGTCGAT1286 
ValIleAsnTyrLeuAlaArgThrTyrAsnArgLysAsnAsnValAsp 
270275280 
GTTACTAAGGATCTTAAGGCTATGGGAAAACTCAAGCGTGAAGTTGAA1334 
ValThrLysAspLeuLysAlaMetGlyLysLeuLysArgGluValGlu 
285290295 
AAAGCCAACGGTACTTTGTCCTCCCAAAAGTCTGTTCGTATCGAGATT1382 
LysAlaAsnGlyThrLeuSerSerGlnLysSerValArgIleGluIle 
300305310 
GAATCTTTCTTTAACGGTCAAGACTTTTCTGAAACTTTATCCCGTGCT1430 
GluSerPhePheAsnGlyGlnAspPheSerGluThrLeuSerArgAla 
315320325330 
AAGTTCGAGGAGATTAAACATGGATCTCTTCAAGAAGACTTTGAGCCT1478 
LysPheGluGluIleLysHisGlySerLeuGlnGluAspPheGluPro 
335340345 
GTTGAGCAAGTATTAAAGGACTCCAACCTCAAGAAATCCGAGATTGAT1526 
ValGluGlnValLeuLysAspSerAsnLeuLysLysSerGluIleAsp 
350355360 
GATATCGTTCTTGTCGGTGGTTCTACTCGTATCCCTAAGGTTCAAGAA1574 
AspIleValLeuValGlyGlySerThrArgIleProLysValGlnGlu 
365370375 
CTTTTGGAGAGCTTCTTTGGTAAGAAGGCTTCTAAGGGTATCAATCCC1622 
LeuLeuGluSerPhePheGlyLysLysAlaSerLysGlyIleAsnPro 
380385390 
GATGAGGCTGTTGCCTATGGTGCTGCTGTTCAAGCCGGCGTTTTATCT1670 
AspGluAlaValAlaTyrGlyAlaAlaValGlnAlaGlyValLeuSer 
395400405410 
GGCGAGGAAGGAAGTGATAACATTGTCCTCTTGGACGTTATCCCTCTT1718 
GlyGluGluGlySerAspAsnIleValLeuLeuAspValIleProLeu 
415420425 
ACTTTAGGTATTGAGACTACCGGTGGTGTTATGACTAAACTTATCGGT1766 
ThrLeuGlyIleGluThrThrGlyGlyValMetThrLysLeuIleGly 
430435440 
CGTAACACTCCTATTCCTACTCGTAAGTCGCAAATTTTCTCTACTGCG1814 
ArgAsnThrProIleProThrArgLysSerGlnIlePheSerThrAla 
445450455 
GTTGACAATCAAAATACTGTTTTAATTCAAGTCTATGAAGGTGAACGT1862 
ValAspAsnGlnAsnThrValLeuIleGlnValTyrGluGlyGluArg 
460465470 
ACTCTTACTAAGGACAACAACCTTCTTGGAAAATTTGACCTTCGTGGT1910 
ThrLeuThrLysAspAsnAsnLeuLeuGlyLysPheAspLeuArgGly 
475480485490 
ATTCCTCCTGCCCCTCGTGGTGTTCCCCAAATTGAAGTCACGTTTGAA1958 
IleProProAlaProArgGlyValProGlnIleGluValThrPheGlu 
495500505 
GTCGATGCCAATGGTGTTTTGACTGTTTCAGCCGTCGACAAGTCTGGT2006 
ValAspAlaAsnGlyValLeuThrValSerAlaValAspLysSerGly 
510515520 
AAGGGTAAGCCTGAGAAGCTTGTTATCAAGAATGACAAAGGTCGTTTG2054 
LysGlyLysProGluLysLeuValIleLysAsnAspLysGlyArgLeu 
525530535 
TCTGAGGAAGATATCGAGCGCATGGTTAAGGAGGCCGAAGAATTCGCT2102 
SerGluGluAspIleGluArgMetValLysGluAlaGluGluPheAla 
540545550 
GAAGAAGATAAGATTTTGAAGGAGCGTATTGAAGCTCGTAATACTCTT2150 
GluGluAspLysIleLeuLysGluArgIleGluAlaArgAsnThrLeu 
555560565570 
GAAAACTACGCCTATTCTTTGAAAGGTCAATTTGACGATGATGAGCAA2198 
GluAsnTyrAlaTyrSerLeuLysGlyGlnPheAspAspAspGluGln 
575580585 
TTAGGTGGTAAGGTTGATCCCGAAGATAAGCAAGCTGTTTTGGACGCT2246 
LeuGlyGlyLysValAspProGluAspLysGlnAlaValLeuAspAla 
590595600 
GTCGAAGATGTTGCTGAATGGCTTGAAATCCACGGAGAAGATGCCAGC2294 
ValGluAspValAlaGluTrpLeuGluIleHisGlyGluAspAlaSer 
605610615 
AAGGAAGAATTTGAAGATCAGCGTCAAAAACTCGATGCCGTTGTTCAT2342 
LysGluGluPheGluAspGlnArgGlnLysLeuAspAlaValValHis 
620625630 
CCTATTACCCAAAAGTTGTATTCCGAAGGAGCTGGTGATGCTGATGAA2390 
ProIleThrGlnLysLeuTyrSerGluGlyAlaGlyAspAlaAspGlu 
635640645650 
GAGGATGATGATTACTTCGATGATGAGGCCGATGAACTTTAAAGTGTTT2439 
GluAspAspAspTyrPheAspAspGluAlaAspGluLeu 
655660 
TAAAATTGCCTGTACTTTCATTTTTTAAGCTTTACTTAGTAATTTTTATTTAGTTCGAAG2499 
TATACGCAAGTCTGACTCGAATGCTCTCATGGTTTCATGACCTTAATCTAAGGGTATTTG2559 
GAAACCAAATGTTTT2574 
(2) INFORMATION FOR SEQ ID NO:7: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 663 amino acids 
(B) TYPE: amino acid 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: 
MetLysLysPheGlnLeuPheSerIleLeuSerTyrPheValAlaLeu 
151015 
PheLeuLeuProMetAlaPheAlaSerGlyAspAspAsnSerThrGlu 
202530 
SerTyrGlyThrValIleGlyIleAspLeuGlyThrThrTyrSerCys 
354045 
ValAlaValMetLysAsnGlyArgValGluIleIleAlaAsnAspGln 
505560 
GlyAsnArgIleThrProSerTyrValAlaPheThrGluAspGluArg 
65707580 
LeuValGlyGluAlaAlaLysAsnGlnAlaProSerAsnProGluAsn 
859095 
ThrIlePheAspIleLysArgLeuIleGlyArgLysPheAspGluLys 
100105110 
ThrMetAlaLysAspIleLysSerPheProPheHisIleValAsnAsp 
115120125 
LysAsnArgProLeuValGluValAsnValGlyGlyLysLysLysLys 
130135140 
PheThrProGluGluIleSerAlaMetIleLeuSerLysMetLysGln 
145150155160 
ThrAlaGluAlaTyrLeuGlyLysProValThrHisSerValValThr 
165170175 
ValProAlaTyrPheAsnAspAlaGlnArgGlnAlaThrLysAspAla 
180185190 
GlyThrIleAlaGlyLeuAsnValIleArgIleValAsnGluProThr 
195200205 
AlaAlaAlaIleAlaTyrGlyLeuAspLysThrAspThrGluLysHis 
210215220 
IleValValTyrAspLeuGlyGlyGlyThrPheAspValSerLeuLeu 
225230235240 
SerIleAspAsnGlyValPheGluValLeuAlaThrSerGlyAspThr 
245250255 
HisLeuGlyGlyGluAspPheAspAsnArgValIleAsnTyrLeuAla 
260265270 
ArgThrTyrAsnArgLysAsnAsnValAspValThrLysAspLeuLys 
275280285 
AlaMetGlyLysLeuLysArgGluValGluLysAlaAsnGlyThrLeu 
290295300 
SerSerGlnLysSerValArgIleGluIleGluSerPhePheAsnGly 
305310315320 
GlnAspPheSerGluThrLeuSerArgAlaLysPheGluGluIleLys 
325330335 
HisGlySerLeuGlnGluAspPheGluProValGluGlnValLeuLys 
340345350 
AspSerAsnLeuLysLysSerGluIleAspAspIleValLeuValGly 
355360365 
GlySerThrArgIleProLysValGlnGluLeuLeuGluSerPhePhe 
370375380 
GlyLysLysAlaSerLysGlyIleAsnProAspGluAlaValAlaTyr 
385390395400 
GlyAlaAlaValGlnAlaGlyValLeuSerGlyGluGluGlySerAsp 
405410415 
AsnIleValLeuLeuAspValIleProLeuThrLeuGlyIleGluThr 
420425430 
ThrGlyGlyValMetThrLysLeuIleGlyArgAsnThrProIlePro 
435440445 
ThrArgLysSerGlnIlePheSerThrAlaValAspAsnGlnAsnThr 
450455460 
ValLeuIleGlnValTyrGluGlyGluArgThrLeuThrLysAspAsn 
465470475480 
AsnLeuLeuGlyLysPheAspLeuArgGlyIleProProAlaProArg 
485490495 
GlyValProGlnIleGluValThrPheGluValAspAlaAsnGlyVal 
500505510 
LeuThrValSerAlaValAspLysSerGlyLysGlyLysProGluLys 
515520525 
LeuValIleLysAsnAspLysGlyArgLeuSerGluGluAspIleGlu 
530535540 
ArgMetValLysGluAlaGluGluPheAlaGluGluAspLysIleLeu 
545550555560 
LysGluArgIleGluAlaArgAsnThrLeuGluAsnTyrAlaTyrSer 
565570575 
LeuLysGlyGlnPheAspAspAspGluGlnLeuGlyGlyLysValAsp 
580585590 
ProGluAspLysGlnAlaValLeuAspAlaValGluAspValAlaGlu 
595600605 
TrpLeuGluIleHisGlyGluAspAlaSerLysGluGluPheGluAsp 
610615620 
GlnArgGlnLysLeuAspAlaValValHisProIleThrGlnLysLeu 
625630635640 
TyrSerGluGlyAlaGlyAspAlaAspGluGluAspAspAspTyrPhe 
645650655 
AspAspGluAlaAspGluLeu 
660 
(2) INFORMATION FOR SEQ ID NO:8: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 6030 base pairs 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: double 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(ix) FEATURE: 
(A) NAME/KEY: CDS 
(B) LOCATION: 1004..4753 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: 
TTTTATCCTATGTCACGGACGACGACTTGTATCACCTTGAATTTTCTGACCAAAGGGGCC60 
GAGTCGCTTCACGAGGGGATGAGAAAGGAAAAGAAGGGAAAACTAAACTTATATAACGCA120 
GGTGTGTCTTTCTACCATTGCCATCAAGTTATTAAAGGCCACGAACAGGAACGCTAGAGA180 
CCTGAGTTTGTCATTTGTTTAGTTCAAGGATTAAATAAACAATCCTTCTACAAATAAGTC240 
CTTTCTTTCACCATCGTCTTAAGACCACTGCCTCCAACGAAAACTAACCTAAAAGAGTTT300 
AGATCACGAGTATTTTCGCTCTTTCCCTCCTTCCCCTGGTTTTTTCTCGTTAGTTCTTTT360 
CATTTAAAAACTCTTCTCTTGTCAAGAATTTAAAAGACGAAGAGTCCAACACCGACTGAT420 
TTTCTAACAGCAAAGGAACGAAGTTTTGCCGTGCAAACAATAATTTCTAAATTATAATTT480 
TGAGCCTAGCTGAGAAATAGGAGAGATTATATTTTAGAAAGGTAAGAAGTTTTTCTGTCA540 
TTCCTTTTAGAATATTTGCTACGTTCTAACATTTTTTGTTACTCAAGCGCATTTTCTGCA600 
ACTTCCCTTATAAGCTATTTCCTTTTTTTGGGACCGATCCTTTCTTCTGTCTTTGGTAAC660 
CTAAAAACCGGAATAGTCAAAGTTATCTGCATAGTCTTCTTGCCAGGCTTATTTTCGCCA720 
TACCATTTTTCTGGTACCCTAAACATTTTGGTCTTATTTTAGAACAGCTGGTGCCTCGTT780 
TTTCCGCATTAGGCGCACTTTTTTCATAGCCACTATTCTAAAAGAAACAACTTTTTTTCA840 
AAGGGAAATCTAAGTTGCCTGCACGAAGAATAAGACAAGGGTTCATAAACGTATAGTATT900 
TGCCAAGTTCCATCTTTTTCTTTGTCACTTTAATATCGCAAAACAGAACACCAAAAACCT960 
TTCAGCGCAAAGATTTGGCCCAATTATTCCATCTTTATACACTATGTCTAAAAAT1015 
MetSerLysAsn 
1 
AGCAACGTTAACAACAATAGATCCCAAGAGCCAAATAACATGTTTGTG1063 
SerAsnValAsnAsnAsnArgSerGlnGluProAsnAsnMetPheVal 
5101520 
CAAACCACAGGAGGTGGTAAAAACGCCCCAAAGCAGATTCATGTTGCA1111 
GlnThrThrGlyGlyGlyLysAsnAlaProLysGlnIleHisValAla 
253035 
CACAGACGTTCCCAAAGTGAGTTGACAAATTTGATGATTGAACAATTC1159 
HisArgArgSerGlnSerGluLeuThrAsnLeuMetIleGluGlnPhe 
404550 
ACTTTGCAGAAGCAGTTGGAGCAAGTTCAAGCACAGCAGCAACAGTTG1207 
ThrLeuGlnLysGlnLeuGluGlnValGlnAlaGlnGlnGlnGlnLeu 
556065 
ATGGCTCAGCAACAGCAATTGGCACAACAGACAGGACAATACCTGTCA1255 
MetAlaGlnGlnGlnGlnLeuAlaGlnGlnThrGlyGlnTyrLeuSer 
707580 
GGAAATTCTGGCTCTAACAATCATTTCACGCCTCAACCGCCTCACCCT1303 
GlyAsnSerGlySerAsnAsnHisPheThrProGlnProProHisPro 
859095100 
CATTACAACTCAAACGGTAATTCACCTGGTATGAGTGCAGGTGGCAGC1351 
HisTyrAsnSerAsnGlyAsnSerProGlyMetSerAlaGlyGlySer 
105110115 
AGAAGTAGAACTCACTCCAGGAACAACTCCGGATATTATCATAATTCA1399 
ArgSerArgThrHisSerArgAsnAsnSerGlyTyrTyrHisAsnSer 
120125130 
TATGATAACAATAACAATAGCAATAATCCTGGGTCTAACTCACACAGA1447 
TyrAspAsnAsnAsnAsnSerAsnAsnProGlySerAsnSerHisArg 
135140145 
AAGACGAGTTCACAATCCAGCATATATGGCCATTCCAGAAGACATTCT1495 
LysThrSerSerGlnSerSerIleTyrGlyHisSerArgArgHisSer 
150155160 
TTAGGTCTAAATGAAGCGAAAAAGGCTGCTGCGGAAGAACAAGCTAAA1543 
LeuGlyLeuAsnGluAlaLysLysAlaAlaAlaGluGluGlnAlaLys 
165170175180 
AGAATATCTGGGGGTGAAGCAGGCGTAACTGTGAAGATAGATTCTGTT1591 
ArgIleSerGlyGlyGluAlaGlyValThrValLysIleAspSerVal 
185190195 
CAAGCTGATAGTGGCTCAAATTCTACTACAGAACAATCTGATTTTAAA1639 
GlnAlaAspSerGlySerAsnSerThrThrGluGlnSerAspPheLys 
200205210 
TTTCCACCACCACCAAATGCTCATCAGGGCCATCGTCGCGCAACTTCA1687 
PheProProProProAsnAlaHisGlnGlyHisArgArgAlaThrSer 
215220225 
AACCTATCACCTCCCTCTTTCAAATTTCCTCCAAACTCTCACGGGGAT1735 
AsnLeuSerProProSerPheLysPheProProAsnSerHisGlyAsp 
230235240 
AATGACGATGAATTCATAGCAACCTCTTCAACGCACCGCCGTTCAAAG1783 
AsnAspAspGluPheIleAlaThrSerSerThrHisArgArgSerLys 
245250255260 
ACAAGAAACAATGAATATTCTCCAGGCATTAATTCCAACTGGAGAAAC1831 
ThrArgAsnAsnGluTyrSerProGlyIleAsnSerAsnTrpArgAsn 
265270275 
CAATCACAGCAACCTCAACAGCAGCTTTCTCCATTCCGCCACAGAGGA1879 
GlnSerGlnGlnProGlnGlnGlnLeuSerProPheArgHisArgGly 
280285290 
TCTAATTCAAGGGATTACAATTCCTTCAATACCTTAGAACCTCCTGCG1927 
SerAsnSerArgAspTyrAsnSerPheAsnThrLeuGluProProAla 
295300305 
ATATTTCAGCAGGGACACAAACATCGTGCCTCTAATTCATCAGTTCAT1975 
IlePheGlnGlnGlyHisLysHisArgAlaSerAsnSerSerValHis 
310315320 
AGTTTCAGTTCACAAGGTAATAATAACGGAGGTGGACGTAAGTCCCTA2023 
SerPheSerSerGlnGlyAsnAsnAsnGlyGlyGlyArgLysSerLeu 
325330335340 
TTTGCACCCTACCTTCCCCAAGCCAACATTCCAGAGCTAATCCAAGAA2071 
PheAlaProTyrLeuProGlnAlaAsnIleProGluLeuIleGlnGlu 
345350355 
GGGAGACTAGTAGCTGGTATATTAAGAGTTAATAAAAAGAATAGATCG2119 
GlyArgLeuValAlaGlyIleLeuArgValAsnLysLysAsnArgSer 
360365370 
GATGCCTGGGTCTCTACAGATGGCGCTCTTGATGCGGATATTTACATT2167 
AspAlaTrpValSerThrAspGlyAlaLeuAspAlaAspIleTyrIle 
375380385 
TGCGGCTCCAAAGATCGTAATAGAGCACTTGAAGGTGATTTAGTCGCG2215 
CysGlySerLysAspArgAsnArgAlaLeuGluGlyAspLeuValAla 
390395400 
GTAGAACTATTAGTTGTGGACGATGTTTGGGAGTCCAAGAAAGAAAAG2263 
ValGluLeuLeuValValAspAspValTrpGluSerLysLysGluLys 
405410415420 
GAAGAAAAGAAGAGGAGAAAGGATGCCTCTATGCAACACGATCTAATT2311 
GluGluLysLysArgArgLysAspAlaSerMetGlnHisAspLeuIle 
425430435 
CCTTTGAACAGTAGTGACGATTACCACAACGATGCATCTGTTACTGCT2359 
ProLeuAsnSerSerAspAspTyrHisAsnAspAlaSerValThrAla 
440445450 
GCAACAAGCAACAATTTTCTATCTTCTCCCTCCTCGTCTGATTCGCTA2407 
AlaThrSerAsnAsnPheLeuSerSerProSerSerSerAspSerLeu 
455460465 
AGCAAGGATGATTTATCCGTCAGAAGAAAGAGGTCATCTACTATCAAT2455 
SerLysAspAspLeuSerValArgArgLysArgSerSerThrIleAsn 
470475480 
AATGATAGTGATTCCTTATCATCTCCTACCAAATCAGGAGTAAGGAGA2503 
AsnAspSerAspSerLeuSerSerProThrLysSerGlyValArgArg 
485490495500 
AGAAGTTCATTGAAACAACGTCCAACTCAAAAGAAAAATGACGATGTT2551 
ArgSerSerLeuLysGlnArgProThrGlnLysLysAsnAspAspVal 
505510515 
GAAGTTGAAGGTCAGTCATTGTTATTAGTTGAAGAAGAAGAAATCAAC2599 
GluValGluGlyGlnSerLeuLeuLeuValGluGluGluGluIleAsn 
520525530 
GATAAATATAAGCCACTTTACGCAGGCCATGTCGTTGCTGTTTTGGAC2647 
AspLysTyrLysProLeuTyrAlaGlyHisValValAlaValLeuAsp 
535540545 
CGTATCCCTGGTCAGTTATTTAGCGGTACATTAGGTTTGTTGAGACCA2695 
ArgIleProGlyGlnLeuPheSerGlyThrLeuGlyLeuLeuArgPro 
550555560 
TCCCAACAAGCTAATAGCGACAATAACAAACCACCACAAAGCCCAAAA2743 
SerGlnGlnAlaAsnSerAspAsnAsnLysProProGlnSerProLys 
565570575580 
ATTGCTTGGTTCAAGCCTACTGATAAGAAGGTGCCATTAATTGCAATT2791 
IleAlaTrpPheLysProThrAspLysLysValProLeuIleAlaIle 
585590595 
CCTACAGAATTAGCTCCAAAGGACTTTGTTGAAAACGCTGATAAATAC2839 
ProThrGluLeuAlaProLysAspPheValGluAsnAlaAspLysTyr 
600605610 
TCCGAAAAGTTATTCGTTGCCTCTATTAAACGTTGGCCAATCACATCT2887 
SerGluLysLeuPheValAlaSerIleLysArgTrpProIleThrSer 
615620625 
TTGCATCCATTTGGTATTTTAGTTTCCGAACTTGGAGATATTCACGAT2935 
LeuHisProPheGlyIleLeuValSerGluLeuGlyAspIleHisAsp 
630635640 
CCTGATACTGAAATTGATTCCATTTTAAGGGATAACAATTTTCTTTCG2983 
ProAspThrGluIleAspSerIleLeuArgAspAsnAsnPheLeuSer 
645650655660 
AATGAATATTTGGATCAAAAAAATCCGCAAAAAGAAAAACCAAGTTTT3031 
AsnGluTyrLeuAspGlnLysAsnProGlnLysGluLysProSerPhe 
665670675 
CAGCCGCTACCATTAACGGCTGAAAGTCTAGAATATAGGAGGAATTTT3079 
GlnProLeuProLeuThrAlaGluSerLeuGluTyrArgArgAsnPhe 
680685690 
ACGGACACTAATGAGTACAATATCTTTGCAATTTCCGAGCTTGGATGG3127 
ThrAspThrAsnGluTyrAsnIlePheAlaIleSerGluLeuGlyTrp 
695700705 
GTGTCTGAATTTGCCTTACATGTCAGGAATAACGGAAATGGTACCCTA3175 
ValSerGluPheAlaLeuHisValArgAsnAsnGlyAsnGlyThrLeu 
710715720 
GAGCTGGGTTGTCATGTTGTTGATGTGACCAGCCATATTGAAGAAGGC3223 
GluLeuGlyCysHisValValAspValThrSerHisIleGluGluGly 
725730735740 
TCCTCTGTTGATAGGCGTGCGAGAAAGAGGTCCTCTGCGGTGTTCATG3271 
SerSerValAspArgArgAlaArgLysArgSerSerAlaValPheMet 
745750755 
CCACAAAAACTTGTCAATTTATTACCACAATCGTTCAACGACGAACTG3319 
ProGlnLysLeuValAsnLeuLeuProGlnSerPheAsnAspGluLeu 
760765770 
TCGTTGGCCCCTGGCAAGGAATCAGCCACGCTGTCGGTTGTTTACACT3367 
SerLeuAlaProGlyLysGluSerAlaThrLeuSerValValTyrThr 
775780785 
CTAGACTCATCTACTTTAAGGATTAAATCTACTTGGGTAGGCGAATCT3415 
LeuAspSerSerThrLeuArgIleLysSerThrTrpValGlyGluSer 
790795800 
ACAATTTCCCCCTCAAACATCTTGTCTTTAGAACAATTAGACGAAAAA3463 
ThrIleSerProSerAsnIleLeuSerLeuGluGlnLeuAspGluLys 
805810815820 
TTATCTACTGGAAGTCCCACTAGCTACCTCTCTACTGTACAGGAAATT3511 
LeuSerThrGlySerProThrSerTyrLeuSerThrValGlnGluIle 
825830835 
GCTAGATCATTTTATGCTAGAAGAATAAATGATCCAGAAGCTACATTA3559 
AlaArgSerPheTyrAlaArgArgIleAsnAspProGluAlaThrLeu 
840845850 
CTTCCCACCCTGTCCTTATTGGAAAGCTTGGATGACGAAAAAGTTAAG3607 
LeuProThrLeuSerLeuLeuGluSerLeuAspAspGluLysValLys 
855860865 
GTTGACTTGAACATCCTGGATAGAACTTTAGGCTTTGTTGTAATTAAT3655 
ValAspLeuAsnIleLeuAspArgThrLeuGlyPheValValIleAsn 
870875880 
GAGATTAAAAGAAAGGTCAACTCCACTGTTGCAGAGAAAATTTACACC3703 
GluIleLysArgLysValAsnSerThrValAlaGluLysIleTyrThr 
885890895900 
AAACTTGGTGATCTAGCTCTTTTGAGAAGGCAGATGCAACCCATTGCA3751 
LysLeuGlyAspLeuAlaLeuLeuArgArgGlnMetGlnProIleAla 
905910915 
ACCAAGATGGCGTCATTTAGAAAGAAAATTCAAAATTTTGGTTACAAT3799 
ThrLysMetAlaSerPheArgLysLysIleGlnAsnPheGlyTyrAsn 
920925930 
TTTGATACCAATACGGCGGATGAATTAATCAAAGGGGTGCTAAAAATT3847 
PheAspThrAsnThrAlaAspGluLeuIleLysGlyValLeuLysIle 
935940945 
AAAGATGACGATGTTAGAGTCGGAATTGAAATTTTACTGTTTAAAACC3895 
LysAspAspAspValArgValGlyIleGluIleLeuLeuPheLysThr 
950955960 
ATGCCAAGAGCTAGATACTTTATTGCTGGCAAAGTAGACCCGGACCAA3943 
MetProArgAlaArgTyrPheIleAlaGlyLysValAspProAspGln 
965970975980 
TATGGGCATTATGCCTTGAACCTACCTATCTACACACATTTCACAGCG3991 
TyrGlyHisTyrAlaLeuAsnLeuProIleTyrThrHisPheThrAla 
985990995 
CCAATGAGAAGATACGCTGATCATGTCGTTCATAGGCAATTAAAGGCC4039 
ProMetArgArgTyrAlaAspHisValValHisArgGlnLeuLysAla 
100010051010 
GTTATCCACGATACTCCATACACCGAAGATATGGAAGCTTTGAAGATT4087 
ValIleHisAspThrProTyrThrGluAspMetGluAlaLeuLysIle 
101510201025 
ACCTCCGAATATTGTAATTTTAAAAAGGACTGTGCTTATCAAGCACAG4135 
ThrSerGluTyrCysAsnPheLysLysAspCysAlaTyrGlnAlaGln 
103010351040 
GAACAAGCAATTCATCTATTGTTGTGTAAAACAATCAACGACATGGGA4183 
GluGlnAlaIleHisLeuLeuLeuCysLysThrIleAsnAspMetGly 
1045105010551060 
AATACTACAGGACAATTATTAACAATGGCTACTGTCTTACAAGTTTAC4231 
AsnThrThrGlyGlnLeuLeuThrMetAlaThrValLeuGlnValTyr 
106510701075 
GAGTCCTCCTTTGATGTATTTATTCCAGAATTTGGTATTGAAAAGAGA4279 
GluSerSerPheAspValPheIleProGluPheGlyIleGluLysArg 
108010851090 
GTTCATGGAGATCAACTACCTTTGATCAAAGCTGAGTTTGATGGTACC4327 
ValHisGlyAspGlnLeuProLeuIleLysAlaGluPheAspGlyThr 
109511001105 
AATCGTGTCTTGGAATTGCATTGGCAGCCCGGCGTAGATAGTGCAACT4375 
AsnArgValLeuGluLeuHisTrpGlnProGlyValAspSerAlaThr 
111011151120 
TTTATACCAGCAGATGAAAAAAATCCAAAATCCTATAGAAATTCCATT4423 
PheIleProAlaAspGluLysAsnProLysSerTyrArgAsnSerIle 
1125113011351140 
AAGAACAAATTCAGATCCACAGCCGCTGAGATTGCGAATATTGAACTA4471 
LysAsnLysPheArgSerThrAlaAlaGluIleAlaAsnIleGluLeu 
114511501155 
GATAAAGAAGCGGAATCTGAACCATTGATCAGCGATCCATTGAGTAAG4519 
AspLysGluAlaGluSerGluProLeuIleSerAspProLeuSerLys 
116011651170 
GAACTCAGCGATTTGCATCTAACAGTACCAAATTTAAGGCTACCATCT4567 
GluLeuSerAspLeuHisLeuThrValProAsnLeuArgLeuProSer 
117511801185 
GCAAGCGACAACAAGCAAAATGCTTTAGAAAAATTCATTTCTACTACT4615 
AlaSerAspAsnLysGlnAsnAlaLeuGluLysPheIleSerThrThr 
119011951200 
GAAACCAGAATTGAAAATGATAACTATATACAAGAAATACATGAATTG4663 
GluThrArgIleGluAsnAspAsnTyrIleGlnGluIleHisGluLeu 
1205121012151220 
CAAAAGATTCCTATTCTATTGAGAGCTGAGGTGGGGATGGCTTTGCCA4711 
GlnLysIleProIleLeuLeuArgAlaGluValGlyMetAlaLeuPro 
122512301235 
TGTTTAACCGTCCGTGCATTAAATCCATTCATGAAGAGGGTA4753 
CysLeuThrValArgAlaLeuAsnProPheMetLysArgVal 
124012451250 
TAATCTCTTCTACCAATATCGTCATTGCTGTTTTTCTTGTTTTTCACTTTCGTTCTTTGG4813 
ATTGTGCTTCACCCCTCAGTATCCCTTCCCTTTGTTTTTATTTCCTGCGAACATTAACAA4873 
CTGCATGAATTTTGTACTTCTCCTTTTAATCCACGTTCCGGTAAGGCATCATCCAAATTT4933 
TTTTATTCGACCTCGTTAAGTCATATATTTTTTCCCAAAAATACATAAAACAATAATGCA4993 
GCCTTCTTTTCAATATTTACAACTTTTCAATTTATATTGTCTTTTGTTATTTATACTCTT5053 
ATATATTAAATTTATTCCGTTACTAAATACCCTTTTGCTGTACAAATATCATCAAAGAGA5113 
AGTACTGAAAGCTTACTTTTTATGCGCTGGGTAATTTTTCCGGAAACAATAACGAAATCA5173 
TCGTCGAGCAATTTTGCTCGTACTTCAGAAACTACTGCGTAAACATTTGAGGTCGTACAA5233 
TAAGTAGATAGAAATAAATAAACCAATTTTTCGTCAGCGTTTAATCTGTAGCCAAAGATT5293 
TGTGGTATTCTCACAGTTTGAATAATATTCAGCTACTTCATCAAGTAGTTTTTTTCAATA5353 
GGAGATTCACGGTTCAATAAGTGCATTGATTATGTTCGACCAATTAGCAGTCTTTACCCC5413 
TCAAGGTCAAGTACTTTACCAATATAACTGTTTAGGAAAAAAGTTTTCTGAAATACAAAT5473 
TAACAGCTTTATATCCCAGCTGATTACTTCCCCAGTAACTAGAAAAGAAAGTGTTGCAAA5533 
CGCAAATACAGACGGATTTGATTTCAATCTTTTAACAATCAACAGCGAACACAAAAATTC5593 
TCCTTCATTTAATGCACTATTTTATTTGAATAAGCAACCAGAATTGTATTTCGTAGTGAC5653 
TTTTGCCGAGCAGACTTTAGAGCTTAATCAAGAAACTCAACAAACACTTGCACTGGTGTT5713 
AAAACTCTGGAACTCATTGCATTTAAGTGAATCCATTCTAAAAAATCGTCAGGGCCAAAA5773 
CGAAAAGAACAAGCATAACTACGTCGATATTCTTCAGGGAATTGAAGACGACCTGAAGAA5833 
ATTTGAGCAATATTTTAGGATAAAATATGAAGAGTCAATAAAACAAGACCATATCAATCC5893 
AGATAATTTTACCAAAAATGGATCAGTACCCCAATCGCATAATAAAAATACCAAGAAAAA5953 
ATTGAGGGATACAAAAGGTAAGAAGCAATCTACAGGAAATGTTGGTAGTGGGTAGTAAAG6013 
TGGGGCCGTGATGGTGG6030 
(2) INFORMATION FOR SEQ ID NO:9: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 1250 amino acids 
(B) TYPE: amino acid 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: 
MetSerLysAsnSerAsnValAsnAsnAsnArgSerGlnGluProAsn 
151015 
AsnMetPheValGlnThrThrGlyGlyGlyLysAsnAlaProLysGln 
202530 
IleHisValAlaHisArgArgSerGlnSerGluLeuThrAsnLeuMet 
354045 
IleGluGlnPheThrLeuGlnLysGlnLeuGluGlnValGlnAlaGln 
505560 
GlnGlnGlnLeuMetAlaGlnGlnGlnGlnLeuAlaGlnGlnThrGly 
65707580 
GlnTyrLeuSerGlyAsnSerGlySerAsnAsnHisPheThrProGln 
859095 
ProProHisProHisTyrAsnSerAsnGlyAsnSerProGlyMetSer 
100105110 
AlaGlyGlySerArgSerArgThrHisSerArgAsnAsnSerGlyTyr 
115120125 
TyrHisAsnSerTyrAspAsnAsnAsnAsnSerAsnAsnProGlySer 
130135140 
AsnSerHisArgLysThrSerSerGlnSerSerIleTyrGlyHisSer 
145150155160 
ArgArgHisSerLeuGlyLeuAsnGluAlaLysLysAlaAlaAlaGlu 
165170175 
GluGlnAlaLysArgIleSerGlyGlyGluAlaGlyValThrValLys 
180185190 
IleAspSerValGlnAlaAspSerGlySerAsnSerThrThrGluGln 
195200205 
SerAspPheLysPheProProProProAsnAlaHisGlnGlyHisArg 
210215220 
ArgAlaThrSerAsnLeuSerProProSerPheLysPheProProAsn 
225230235240 
SerHisGlyAspAsnAspAspGluPheIleAlaThrSerSerThrHis 
245250255 
ArgArgSerLysThrArgAsnAsnGluTyrSerProGlyIleAsnSer 
260265270 
AsnTrpArgAsnGlnSerGlnGlnProGlnGlnGlnLeuSerProPhe 
275280285 
ArgHisArgGlySerAsnSerArgAspTyrAsnSerPheAsnThrLeu 
290295300 
GluProProAlaIlePheGlnGlnGlyHisLysHisArgAlaSerAsn 
305310315320 
SerSerValHisSerPheSerSerGlnGlyAsnAsnAsnGlyGlyGly 
325330335 
ArgLysSerLeuPheAlaProTyrLeuProGlnAlaAsnIleProGlu 
340345350 
LeuIleGlnGluGlyArgLeuValAlaGlyIleLeuArgValAsnLys 
355360365 
LysAsnArgSerAspAlaTrpValSerThrAspGlyAlaLeuAspAla 
370375380 
AspIleTyrIleCysGlySerLysAspArgAsnArgAlaLeuGluGly 
385390395400 
AspLeuValAlaValGluLeuLeuValValAspAspValTrpGluSer 
405410415 
LysLysGluLysGluGluLysLysArgArgLysAspAlaSerMetGln 
420425430 
HisAspLeuIleProLeuAsnSerSerAspAspTyrHisAsnAspAla 
435440445 
SerValThrAlaAlaThrSerAsnAsnPheLeuSerSerProSerSer 
450455460 
SerAspSerLeuSerLysAspAspLeuSerValArgArgLysArgSer 
465470475480 
SerThrIleAsnAsnAspSerAspSerLeuSerSerProThrLysSer 
485490495 
GlyValArgArgArgSerSerLeuLysGlnArgProThrGlnLysLys 
500505510 
AsnAspAspValGluValGluGlyGlnSerLeuLeuLeuValGluGlu 
515520525 
GluGluIleAsnAspLysTyrLysProLeuTyrAlaGlyHisValVal 
530535540 
AlaValLeuAspArgIleProGlyGlnLeuPheSerGlyThrLeuGly 
545550555560 
LeuLeuArgProSerGlnGlnAlaAsnSerAspAsnAsnLysProPro 
565570575 
GlnSerProLysIleAlaTrpPheLysProThrAspLysLysValPro 
580585590 
LeuIleAlaIleProThrGluLeuAlaProLysAspPheValGluAsn 
595600605 
AlaAspLysTyrSerGluLysLeuPheValAlaSerIleLysArgTrp 
610615620 
ProIleThrSerLeuHisProPheGlyIleLeuValSerGluLeuGly 
625630635640 
AspIleHisAspProAspThrGluIleAspSerIleLeuArgAspAsn 
645650655 
AsnPheLeuSerAsnGluTyrLeuAspGlnLysAsnProGlnLysGlu 
660665670 
LysProSerPheGlnProLeuProLeuThrAlaGluSerLeuGluTyr 
675680685 
ArgArgAsnPheThrAspThrAsnGluTyrAsnIlePheAlaIleSer 
690695700 
GluLeuGlyTrpValSerGluPheAlaLeuHisValArgAsnAsnGly 
705710715720 
AsnGlyThrLeuGluLeuGlyCysHisValValAspValThrSerHis 
725730735 
IleGluGluGlySerSerValAspArgArgAlaArgLysArgSerSer 
740745750 
AlaValPheMetProGlnLysLeuValAsnLeuLeuProGlnSerPhe 
755760765 
AsnAspGluLeuSerLeuAlaProGlyLysGluSerAlaThrLeuSer 
770775780 
ValValTyrThrLeuAspSerSerThrLeuArgIleLysSerThrTrp 
785790795800 
ValGlyGluSerThrIleSerProSerAsnIleLeuSerLeuGluGln 
805810815 
LeuAspGluLysLeuSerThrGlySerProThrSerTyrLeuSerThr 
820825830 
ValGlnGluIleAlaArgSerPheTyrAlaArgArgIleAsnAspPro 
835840845 
GluAlaThrLeuLeuProThrLeuSerLeuLeuGluSerLeuAspAsp 
850855860 
GluLysValLysValAspLeuAsnIleLeuAspArgThrLeuGlyPhe 
865870875880 
ValValIleAsnGluIleLysArgLysValAsnSerThrValAlaGlu 
885890895 
LysIleTyrThrLysLeuGlyAspLeuAlaLeuLeuArgArgGlnMet 
900905910 
GlnProIleAlaThrLysMetAlaSerPheArgLysLysIleGlnAsn 
915920925 
PheGlyTyrAsnPheAspThrAsnThrAlaAspGluLeuIleLysGly 
930935940 
ValLeuLysIleLysAspAspAspValArgValGlyIleGluIleLeu 
945950955960 
LeuPheLysThrMetProArgAlaArgTyrPheIleAlaGlyLysVal 
965970975 
AspProAspGlnTyrGlyHisTyrAlaLeuAsnLeuProIleTyrThr 
980985990 
HisPheThrAlaProMetArgArgTyrAlaAspHisValValHisArg 
99510001005 
GlnLeuLysAlaValIleHisAspThrProTyrThrGluAspMetGlu 
101010151020 
AlaLeuLysIleThrSerGluTyrCysAsnPheLysLysAspCysAla 
1025103010351040 
TyrGlnAlaGlnGluGlnAlaIleHisLeuLeuLeuCysLysThrIle 
104510501055 
AsnAspMetGlyAsnThrThrGlyGlnLeuLeuThrMetAlaThrVal 
106010651070 
LeuGlnValTyrGluSerSerPheAspValPheIleProGluPheGly 
107510801085 
IleGluLysArgValHisGlyAspGlnLeuProLeuIleLysAlaGlu 
109010951100 
PheAspGlyThrAsnArgValLeuGluLeuHisTrpGlnProGlyVal 
1105111011151120 
AspSerAlaThrPheIleProAlaAspGluLysAsnProLysSerTyr 
112511301135 
ArgAsnSerIleLysAsnLysPheArgSerThrAlaAlaGluIleAla 
114011451150 
AsnIleGluLeuAspLysGluAlaGluSerGluProLeuIleSerAsp 
115511601165 
ProLeuSerLysGluLeuSerAspLeuHisLeuThrValProAsnLeu 
117011751180 
ArgLeuProSerAlaSerAspAsnLysGlnAsnAlaLeuGluLysPhe 
1185119011951200 
IleSerThrThrGluThrArgIleGluAsnAspAsnTyrIleGlnGlu 
120512101215 
IleHisGluLeuGlnLysIleProIleLeuLeuArgAlaGluValGly 
122012251230 
MetAlaLeuProCysLeuThrValArgAlaLeuAsnProPheMetLys 
123512401245 
ArgVal 
1250 
(2) INFORMATION FOR SEQ ID NO:10: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 168 amino acids 
(B) TYPE: amino acid 
(C) STRANDEDNESS: single 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: 
IleProProAlaProArgGlyValProGlnIleGluValThrPheGlu 
151015 
IleAspValAsnGlyIleLeuArgValThrAlaGluAspLysGlyThr 
202530 
GlyAsnLysAsnLysIleThrIleThrAsnAspGlnAsnArgLeuThr 
354045 
ProGluGluIleGluArgMetValAsnAspAlaGluLysPheAlaGlu 
505560 
GluAspLysLysLeuLysGluArgIleAspThrArgAsnGluLeuGlu 
65707580 
SerTyrAlaTyrSerLeuLysAsnGlnIleGlyAspLysGluLysLeu 
859095 
GlyGlyLysLeuSerSerGluGlyLysGluThrMetGluLysAlaVal 
100105110 
GluGluLysIleGluTrpLeuGluSerHisGlnAspAlaAspIleGlu 
115120125 
AspPheLysAlaLysLysLysGluLeuGluGluIleValGlnProIle 
130135140 
IleSerLysLeuTyrGlySerGlyGlyProProProThrGlyGluGlu 
145150155160 
AspThrSerGluLysAspGluLeu 
165 
(2) INFORMATION FOR SEQ ID NO:11: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 654 amino acids 
(B) TYPE: amino acid 
(C) STRANDEDNESS: single 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: 
MetLysPheProMetValAlaAlaAlaLeuLeuLeuLeuCysAlaVal 
151015 
ArgAlaGluGluGluAspLysLysGluAspValGlyThrValValGly 
202530 
IleAspLeuGlyThrThrTyrSerCysValGlyValPheLysAsnGly 
354045 
ArgValGluIleIleAlaAsnAspGlnGlyAsnArgIleThrProSer 
505560 
TyrValAlaPheThrProGluGlyGluArgLeuIleGlyAspAlaAla 
65707580 
LysAsnGlnLeuThrSerAsnProGluAsnThrValPheAspAlaLys 
859095 
ArgLeuIleGlyArgThrTrpAsnAspProSerValGlnGlnAspIle 
100105110 
LysPheLeuProPheLysValValGluLysLysThrLysProTyrIle 
115120125 
GlnValAspIleGlyGlyGlyGlnThrLysThrPheAlaProGluGlu 
130135140 
IleSerAlaMetValLeuThrLysMetLysGluThrAlaGluAlaTyr 
145150155160 
LeuGlyLysLysValThrHisAlaValValThrValProAlaTyrPhe 
165170175 
AsnAspAlaGlnArgGlnAlaThrLysAspAlaGlyThrIleAlaGly 
180185190 
LeuAsnValMetArgIleIleAsnGluProThrAlaAlaAlaIleAla 
195200205 
TyrGlyLeuAspLysArgGluGlyGluLysAsnIleLeuValPheAsp 
210215220 
LeuGlyGlyGlyThrPheAspValSerLeuLeuThrIleAspAsnGly 
225230235240 
ValPheGluValValAlaThrAsnGlyAspThrHisLeuGlyGlyGlu 
245250255 
AspPheAspGlnArgValMetGluHisPheIleLysLeuTyrLysLys 
260265270 
LysThrGlyLysAspValArgLysAspAsnArgAlaValGlnLysLeu 
275280285 
ArgArgGluValGluLysAlaLysArgAlaLeuSerSerGlnHisGln 
290295300 
AlaArgIleGluIleGluSerPhePheGluGlyGluAspPheSerGlu 
305310315320 
ThrLeuThrArgAlaLysPheGluGluLeuAsnMetAspLeuPheArg 
325330335 
SerThrMetLysProValGlnLysValLeuGluAspSerAspLeuLys 
340345350 
LysSerAspIleAspGluIleValLeuValGlyGlySerThrArgIle 
355360365 
ProLysIleGlnGlnLeuValLysGluPhePheAsnGlyLysGluPro 
370375380 
SerArgGlyIleAsnProAspGluAlaValAlaTyrGlyAlaAlaVal 
385390395400 
GlnAlaGlyValLeuSerGlyAspGlnAspThrGlyAspLeuValLeu 
405410415 
LeuAspValCysProLeuThrLeuGlyIleGluThrValGlyGlyVal 
420425430 
MetThrLysLeuIleProArgAsnThrValValProThrLysLysSer 
435440445 
GlnIlePheSerThrAlaSerAspAsnGlnProThrValThrIleLys 
450455460 
ValTyrGluGlyGluArgProLeuThrLysAspAsnHisLeuLeuGly 
465470475480 
ThrPheAspLeuThrGlyIleProProAlaProArgGlyValProGln 
485490495 
IleGluValThrPheGluIleAspValAsnGlyIleLeuArgValThr 
500505510 
AlaGluAspLysGlyThrGlyAsnLysAsnLysIleThrIleThrAsn 
515520525 
AspGlnAsnArgLeuThrProGluGluIleGluArgMetValAsnAsp 
530535540 
AlaGluLysPheAlaGluGluAspLysLysLeuLysGluArgIleAsp 
545550555560 
ThrArgAsnGluLeuGluSerTyrAlaTyrSerLeuLysAsnGlnIle 
565570575 
GlyAspLysGluLysLeuGlyGlyLysLeuSerSerGluAspLysGlu 
580585590 
ThrMetGluLysAlaValGluGluLysIleGluTrpLeuGluSerHis 
595600605 
GlnAspAlaAspIleGluAspPheLysAlaLysLysLysGluLeuGlu 
610615620 
GluIleValGlnProIleIleSerLysLeuTyrGlySerAlaGlyPro 
625630635640 
ProProThrGlyGluGluAspThrSerGluLysAspGluLeu 
645650 
(2) INFORMATION FOR SEQ ID NO:12: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 5470 base pairs 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: double 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 593..715 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 806..1036 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 1402..1539 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 2175..2289 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 2378..2764 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 2878..3115 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 3400..3568 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 4535..5095 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: 
CCCGGGGTCACTCCTGCTGGACCTACTCCGACCCCCTAGGCCGGGAGTGAAGGCGGGACT60 
TGTGCGGTTACCAGCGGAAATGCCTCGGGGTCAGAAGTCGCAGGAGAGATAGACAGCTGC120 
TGAACCAATGGGACCAGCGGATGGGGCGGATGTTATCTACCATTGGTGAACGTTAGAAAC180 
GAATAGCAGCCAATGAATCAGCTGGGGGGGCGGAGCAGTGACGTTTATTGCGGAGGGGGC240 
CGCTTCGAATCGGCGGCGGCCAGCTTGGTGGCCTGGGCCAATGAACGGCCTCCAACGAGC300 
AGGGCCTTCACCAATCGGCGGCCTCCACGACGGGGCTGGGGGAGGGTATATAAGCCGAGT360 
AGGCGACGGTGAGGTCGACGCCGGCCAAGACAGCACAGACAGATTGACCTATTGGGGTGT420 
TTCGCGAGTGTGAGAGGGAAGCGCCGCGGCCTGTATTTCTAGACCTGCCCTTCGCCTGGT480 
TCGTGGCGCCTTGTGACCCCGGGCCCCTGCCGCCTGCAAGTCGAAATTGCGCTGTGCTCC540 
TGTGCTACGGCCTGTGGCTGGACTGCCTGCTGCTGCCCAACTGGCTGGCAAGATGAAGCT600 
CTCCCTGGTGGCCGCGATGCTGCTGCTGCTCAGCGCGGCGCGGGCCGAGGAGGAGGACAA660 
GAAGGAGGACGTGGGCACGGTGGTCGGCATCGACTTGGGGACCACCTACTCCTGGTAAGT720 
GGGGTTGCGGATGAGGGGGACGGGGCGTGGCGCTGGCTGGCGTGAGAAGTGCGGTGCTGA780 
TGTCCCTCTGTCGGGTTTTTGCAGCGTCGGCGTGTTCAAGAACGGCCGCGTGGAGATCAT840 
CGCCAACGATCAGGGCAACCGCATCACGCCGTCCTATGTCGCCTTCACTCCTGAAGGGGA900 
ACGTCTGATTGGCGATGCCGCCAAGAACCAGCTCACCTCCAACCCCGAGAACACGGTCTT960 
TGACGCCAAGCGGCTCATCGGCCGCACGTGGAATGACCCGTCTGTGCAGCAGGACATCAA1020 
GTTCTTGCCGTTCAAGGTTCGACCGGTTTTCCTCATCCAGTTAGAGAACGGGTGGGTGGT1080 
GGGAGTATTTAGAGTTATAAGTCTCTGGAAAAGTGTTGAGACAACAGTTGAAGGTTATAG1140 
ACATGATGTATGTAATAACTTTAATACTATTAGTATGTTACAAAACTTAAGACAGTTGCT1200 
GTCGTACTGTCTACGATAGTTTAGGAATAAAAGACCGATTAAAACTGAACTTTGTAAGAC1260 
ACCTATACTCCCTGAAGTATTTCTAGTCAATTTGCAGCCCCAAGGGACCAAAATAAACCA1320 
AATTGTGGGGATGGTAGTGGGTCTTTTAAACTTTGAGATGTCATTGTATCTGTGTCTGAA1380 
AACAATAATTCTTTAAAATAGGTGGTTGAAAAGAAAACTAAACCATACATTCAAGTTGAT1440 
ATTGGAGGTGGGCAAACAAAGACATTTGCTCCTGAAGAAATTTCTGCCATGGTTCTCACT1500 
AAAATGAAAGAAACCGCTGAGGCTTATTTGGGAAAGAAGGTAAATATTTCTAGAACAATG1560 
TTAAGTATTTTTTGATCATTAGTATTCTCGGTTGGCTGTTATGTATAGAAGCCTTCGTGA1620 
AGGGTTTCAAAAATTTTAATCAGAATGGTATTCATGCTTGTCACGGTTTAATTATTGAGT1680 
CCCTTTACTATAAGCCAAACAAAAATAGACTTTTCATGTATTATTTAATGCTTACAATTC1740 
CAGGAACAATAAAATTTTATATGTTGTATTCATCAATAATTGGCTTAAAAACTAAAGTGA1800 
TGGTTTGACTGTAATTTTTTTTTTTTGAGATGGAGTCTTGCTCTGTTGCCCAGGCTGGAC1860 
TGCAGTGGCACGATCTCAGCTCACTGCAACCTCTGCCTCCCGGGTTAAGCAGCTCTCCTG1920 
CCTCAGCCTCCAAGTAATGGAACGACAGGCACACCACCACAGCTGGCTAATTTTTTTTTT1980 
TTTTTTTAATTTTCAGTAGAGACAGGGTTTCTCCACATTGCCAGGCTGGTCTTGAAATCC2040 
TGCCCTCAGGTTGATCCTCCTGCCTAGCCTCCCAAAGTGCTGGATTATAGGCAGAAGCCA2100 
CCGCCTGGCCAGACTGTAATTTAAATAAGGGTTAAACTATGTGACAATACACTTAATTAT2160 
CTTTATCCTTTTAGGTTACCCATGCAGTTGTTACTGTACCAGCCTATTTTAATGATGCCC2220 
AACGCCAAGCAACCAAAGACGCTGGAACTATTGCTGGCCTAAATGTTATGAGGATCATCA2280 
ACGAGCCGTAAGTATGAAATTCAGGGATACGGCATATTTGCCAAATAGTGGAAATGTGAA2340 
GTACTGACAAAACTTTTCCCTTTTTCAATCTAATAGTACGGCAGCTGCTATTGCTTATGG2400 
CCTGGATAAGAGGGAGGGGGAGAAGAACATCCTGGTGTTTGACCTGGGTGGCGGAACCTT2460 
CGATGTGTCTCTTCTCACCATTGACAATGGTGTCTTCGAAGTTGTGGCCACTAATGGAGA2520 
TACTCATCTGGGTGGAGAAGACTTTGACCAGCGTGTCATGGAACACTTCATCAAACTGTA2580 
CAAAAAGAAGACGGGCAAAGATGTCAGGAAGGACAATAGAGCTGTGCAGAAACTCCGGCG2640 
CGAGGTAGAAAAGGCCAAGGCCCTGTCTTCTCAGCATCAAGCAAGAATTGAAATTGAGTC2700 
CTTCTATGAAGGAGAAGACTTTTCTGAGACCCTGACTCGGGCCAAATTTGAAGAGCTCAA2760 
CATGGTATGTTCCTTGTTTTCTGCTTTGCTAATGAGATCTCCTTAGACTCTGAATTCAGG2820 
ACATTGCATCTAGATACTTAGATAACAGACATCACAGTAACCATGTCTTTTTTCTAGGAT2880 
CTGTTCCGGTCTACTATGAAGCCCGTCCAGAAAGTGTTGGAAGATTCTGATTTGAAGAAG2940 
TCTGATATTGATGAAATTGTTCTTGTTGGTGGCTCGACTCGAATTCCAAAGATTCAGCAA3000 
CTGGTTAAAGAGTTCTTCAATGGCAAGGAACCATCCCGTGGCATAAACCCAGATGAAGCT3060 
GTAGCGTATGGTGCTGCTGTCCAGGCTGGTGTGCTCTCTGGTGATCAAGATACAGGTAGG3120 
TCATCATCGCAGCATCTTTCTTAGTGATTCAGTAGCTTGATGGAAGAGCTCGGTACCCCT3180 
ATTGCTTTAGAAAATACCAGAATATGAGCAACAAGGTCACACAGCTAGTAAAGGGTATAA3240 
GTGAAGACAAGACTGGGGTAGTCTCCAAGATCATTAGCAACTGTTTAATTCACTGCCTTT3300 
AAAATGTGTGTGTTAGAACCTAACCAAATGTTAGAGAGATAAACTTTACATAGCTCATAG3360 
GGAGAACTTGAATTAAAAGTTAAATAACTTATCCTTACAGGTGACCTGGTACTGCTTCAT3420 
GTATGTCCCCTTACACTTGGTATTGAAACTGTAGGAGGTGTCATGACCAAACTGATTCCA3480 
AGTAATACAGTGGTGCCTACCAAGAACTCTCAGATCTTTTCTACAGCTTCTGATAATCAA3540 
CCAACTGTTACAATCAAGGTCTATGAAGGTAATTACCTTAAGTTTGGTTAATATCATGGC3600 
TTTTTTTTTGAGATGAAGTCTTGCTCTGTTGCCCAGGCTGGACTGCAGTGGCACGATCTC3660 
GGCTCACTGCAAATTCTGTCTCCCGGGTTCAAGTGATTCTCCTGCCTCAGCCTCCAGAGT3720 
AGCTGGATTACAGCCTGACCACCACACCTGGCTAATTTCTGTATTTTTAGTAGAGGATGG3780 
GCTTTCACCATGTTTCCCAGGCTGGTCTCCAACTCCTGACCTCAGGTCATCTGCCTGCCT3840 
CCACCGTCCCGAAAGTACTGGGATTATAGCGTGAGCCACCACGCCAGATCTATCTATCAT3900 
GGCATATTTTAAAAGAACATGACTTAATATGTCCTATTGAAATGGCTAGGGAACTAAGTA3960 
ACTGCTGTTTTCAGATGGAGGTCTTAATTTGAATAATGTTGATATTAGATATTTAGCATT4020 
CTTTTTTTTTTTTTTTTAATGGAGTCTTGCTCTGTCGCCTAGGCTGGGGTGCAGTGGCAT4080 
GACTTGCAACCTCTGCCTCCCGAATAGCTGGGATTACAGGTGCCCACCATCACGCCCGGC4140 
TAAGTTTTGTATTTTTAGTAGAGGCGAGTTTCGCCATGTTGGCCAGGCTGGTCTTGAACC4200 
CCTAACCTCAGTGATCCCACGGTCACCGACCTGGCCTCCCAAAAGTACTGTACCCAGCCA4260 
ATGATTAGCATTCTCACTAATAATAGCATCTGAGCTGGCTCCTAGAGTACAAGAAAAAGG4320 
AGTTCACAGTACTTTAAAATAGATAAAATTCAGTTGAGTTAGTAACCTAACTCATTGTTA4380 
GTACTAGTTGCTGCTCCTTGTAGACCAATATGAAATTACTTTTAGCTCGATAAAACCAAA4440 
AGTGTCACTTTATGCTTCAGACTGAAATGCGGGGATCTAGATGTGCTAATGCTTGTCAGT4500 
AACAACTAACAAGTTTTTCTGTATGTAACTTCTAGGTGAAAGACCCCTGACAAAAGACAA4560 
TCATCTTCTGGGTACATTTGATCTGACTGGAATTCCTCCTGCTCCTCGTGGGGTCCCACA4620 
GATTGAAGTCACCTTTGAGATAGATGTGAATGGTATTCTTCGAGTGACAGCTGAAGACAA4680 
GGGTACAGGGAACAAAAATAAGATCACAATCACCAATGACCAGAATCGCCTGACACCTGA4740 
AGAAATCGAAAGGATGGTTAATGATGCTGAGAAGTTTGCTGAGGAAGACAAAAAGCTGAA4800 
GGAGCGCATTGATACTAGAAATGAGTTGGAAAGCTATGCCTATTCTCTAAAGAATCAGAT4860 
TGGAGATAAAGAAAAGCTGGGAGGTAAACTTTCCTCTGAAGATAAGGAGACCATGGAAAA4920 
AGCTGTAGAAGAAAAGATTGAATGGCTGGAAAGCCACCAAGATGCTGACATTGAAGACTT4980 
CAAAGCTAAGAAGAAGGAACTGGAAGAAATTGTTCAACCAATTATCAGCAAACTCTATGG5040 
AAGTGCAGGCCCTCCCCCAACTGGTGAAGAGGATACAGCAGAAAAAGATGAGTTGTAGAC5100 
ACTGATCTGCTAGTGCTGTAATATTGTAAATACTGGACTCAGGAACTTTTGTTAGGAAAA5160 
AATTGAAAGAACTTAAGTCTCGAATGTAATTGGAATCTTCACCTCAGAGTGGAGTTGAAA5220 
CTGCTATAGCCTAAGCGGCTGTTTACTGCTTTTCATTAGCAGTTGCTCACATGTCTTTGG5280 
GTGGGGGGGAGAAGAAGAATTGGCCATCTTAAAAAGCGGGTAAAAAACCTGGGTTAGGGT5340 
GTGTGTTCACCTTCAAAATGTTCTATTTAACAACTGGGTCATGTGCATCTGGTGTAGGAG5400 
GTTTTTTCTACCATAAGTGACACCAATAAATGTTTGTTATTTACACTGGTCTAATGTTTG5460 
TGAGAAGCTT5470 
(2) INFORMATION FOR SEQ ID NO:13: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 2089 base pairs 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: double 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(ix) FEATURE: 
(A) NAME/KEY: CDS 
(B) LOCATION: 66..2005 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: 
GAGGCAGCTGCCGGGCATTAGTGTGGTCTCGTCGTCAGCGCAGCTGGGCCTACACACAAG60 
CAACCATGTCTAAGGGACCTGCAGTTGGCATTGATCTCGGCACCACC107 
MetSerLysGlyProAlaValGlyIleAspLeuGlyThrThr 
1510 
TACTCCTGTGTGGGTGTCTTCCAGCATGGAAAGGTGGAAATTATTGCC155 
TyrSerCysValGlyValPheGlnHisGlyLysValGluIleIleAla 
15202530 
AATGACCAGGGTAACCGCACCACGCCAAGCTATGTTGCTTTCACGGAC203 
AsnAspGlnGlyAsnArgThrThrProSerTyrValAlaPheThrAsp 
354045 
ACAGAGAGATTAATTGGGGATGCGGCCAAGAATCAGGTTGCAATGAAC251 
ThrGluArgLeuIleGlyAspAlaAlaLysAsnGlnValAlaMetAsn 
505560 
CCCACCAACACAGTTTTTGATGCCAAACGTCTGATCGGGCGTAGGTTT299 
ProThrAsnThrValPheAspAlaLysArgLeuIleGlyArgArgPhe 
657075 
GATGATGCTGTTGTTCAGTCTGATATGAAGCACTGGCCCTTCATGGTG347 
AspAspAlaValValGlnSerAspMetLysHisTrpProPheMetVal 
808590 
GTGAATGATGCAGGCAGGCCCAAGGTCCAAGTCGAATACAAAGGGGAG395 
ValAsnAspAlaGlyArgProLysValGlnValGluTyrLysGlyGlu 
95100105110 
ACAAAAAGTTTCTACCCAGAGGAAGTGTCCTCCATGGTTCTGACAAAG443 
ThrLysSerPheTyrProGluGluValSerSerMetValLeuThrLys 
115120125 
ATGAAGGAAATTGCAGAAGCATACCTCGGAAAGACTGTTACCAACGCT491 
MetLysGluIleAlaGluAlaTyrLeuGlyLysThrValThrAsnAla 
130135140 
GTGGTCACAGTGCCCGCTTACTTCAATGACTCTCAGCGACAGGCAACA539 
ValValThrValProAlaTyrPheAsnAspSerGlnArgGlnAlaThr 
145150155 
AAAGATGCTGGAACTATTGCTGGCCTCAATGTACTTCGAATCATCAAT587 
LysAspAlaGlyThrIleAlaGlyLeuAsnValLeuArgIleIleAsn 
160165170 
GAACCAACTGCTGCTGCTATTGCTTATGGCTTAGATAAGAAGGTCGGA635 
GluProThrAlaAlaAlaIleAlaTyrGlyLeuAspLysLysValGly 
175180185190 
GCTGAAAGGAATGTGCTCATTTTTGACTTGGGAGGTGGCACTTTTGAT683 
AlaGluArgAsnValLeuIlePheAspLeuGlyGlyGlyThrPheAsp 
195200205 
GTGTCAATCCTCACTATTGAGGATGGAATTTTTGAGGTCAAATCAACA731 
ValSerIleLeuThrIleGluAspGlyIlePheGluValLysSerThr 
210215220 
GCTGGAGACACCCACTTAGGCGGAGAAGACTTTGATAACCGAATGGTC779 
AlaGlyAspThrHisLeuGlyGlyGluAspPheAspAsnArgMetVal 
225230235 
AATCATTTCATTGCTGAGTTCAAGCGAAAGCACAAGAAAGACATCAGT827 
AsnHisPheIleAlaGluPheLysArgLysHisLysLysAspIleSer 
240245250 
GAGAACAAGAGAGCTGTCCGCCGTCTCCGCACGGCCTGCGAGCGGGCC875 
GluAsnLysArgAlaValArgArgLeuArgThrAlaCysGluArgAla 
255260265270 
AAGCGCACCCTCTCCTCCAGCACCCAGGCCAGTATTGAGATTGATTCT923 
LysArgThrLeuSerSerSerThrGlnAlaSerIleGluIleAspSer 
275280285 
CTCTATGAGGGAATTGACTTCTATACCTCCATTACCCGTGCTCGATTT971 
LeuTyrGluGlyIleAspPheTyrThrSerIleThrArgAlaArgPhe 
290295300 
GAGGAGTTGAATGCTGACCTGTTCCGTGGCACACTGGACCCTGTAGAG1019 
GluGluLeuAsnAlaAspLeuPheArgGlyThrLeuAspProValGlu 
305310315 
AAGGCCCTTCGAGATGCCAAGCTGGACAAGTCACAGATCCATGATATT1067 
LysAlaLeuArgAspAlaLysLeuAspLysSerGlnIleHisAspIle 
320325330 
GTCTTGGTGGGTGGTTCTACCAGAATCCCCAAGATCCAGAAACTTCTG1115 
ValLeuValGlyGlySerThrArgIleProLysIleGlnLysLeuLeu 
335340345350 
CAAGACTTCTTCAATGGAAAAGAGCTGAACAAGAGCATTAACCCCGAT1163 
GlnAspPhePheAsnGlyLysGluLeuAsnLysSerIleAsnProAsp 
355360365 
GAAGCTGTTGCCTATGGTGCAGCTGTCCAGGCAGCCATTCTATCTGGA1211 
GluAlaValAlaTyrGlyAlaAlaValGlnAlaAlaIleLeuSerGly 
370375380 
GACAAGTCTGAGAACGTTCAGGATTTGCTGCTCTTGGATGTCACTCCT1259 
AspLysSerGluAsnValGlnAspLeuLeuLeuLeuAspValThrPro 
385390395 
CTTTCCCTTGGTATTGAAACTGCTGGCGGAGTCATGACTGTCCTCATC1307 
LeuSerLeuGlyIleGluThrAlaGlyGlyValMetThrValLeuIle 
400405410 
AAGCGCAATACCACCATCCCCACCAAGCAGACACAGACTCTCACCACC1355 
LysArgAsnThrThrIleProThrLysGlnThrGlnThrLeuThrThr 
415420425430 
TACTCTGACAACCAGCCTGGTGTACTCATTCAGGTGTATGAAGGTGAA1403 
TyrSerAspAsnGlnProGlyValLeuIleGlnValTyrGluGlyGlu 
435440445 
AGGGCCATGACCAAGGACAACAACCTGCTTGGAAAGTTCGAGCTCACA1451 
ArgAlaMetThrLysAspAsnAsnLeuLeuGlyLysPheGluLeuThr 
450455460 
GGCATCCCTCCAGCACCCCGTGGGGTTCCTCAGATTGAGGTTACTTTT1499 
GlyIleProProAlaProArgGlyValProGlnIleGluValThrPhe 
465470475 
GACATCGATGCCAATGGCATCCTCAATGTTTCTGCTGTAGATAAGAGC1547 
AspIleAspAlaAsnGlyIleLeuAsnValSerAlaValAspLysSer 
480485490 
ACAGGAAAGGAGAACAAGATCACCATCACCAATGACAAGGGCCGCTTG1595 
ThrGlyLysGluAsnLysIleThrIleThrAsnAspLysGlyArgLeu 
495500505510 
AGTAAGGAAGATATTGAGCGCATGGTCCAAGAAGCTGAGAAGTACAAG1643 
SerLysGluAspIleGluArgMetValGlnGluAlaGluLysTyrLys 
515520525 
GCTGAGGATGAGAAGCAGAGAGATAAGGTTTCCTCCAAGAACTCACTG1691 
AlaGluAspGluLysGlnArgAspLysValSerSerLysAsnSerLeu 
530535540 
GAGTCCTATGCCTTCAACATGAAAGCAACTGTGGAAGATGAGAAACTT1739 
GluSerTyrAlaPheAsnMetLysAlaThrValGluAspGluLysLeu 
545550555 
CAAGGCAAGATCAATGATGAGGACAAACAGAAGATTCTTGACAAGTGC1787 
GlnGlyLysIleAsnAspGluAspLysGlnLysIleLeuAspLysCys 
560565570 
AATGAAATCATCAGCTGGCTGGATAAGAACCAGACTGCAGAGAAGGAA1835 
AsnGluIleIleSerTrpLeuAspLysAsnGlnThrAlaGluLysGlu 
575580585590 
GAATTTGAGCATCAGCAGAAAGAACTGGAGAAAGTCTGCAACCCTATT1883 
GluPheGluHisGlnGlnLysGluLeuGluLysValCysAsnProIle 
595600605 
ATCACCAAGCTGTACCAGAGTGCAGGTGGCATGCCTGGAGGGATGCCT1931 
IleThrLysLeuTyrGlnSerAlaGlyGlyMetProGlyGlyMetPro 
610615620 
GGTGGCTTCCCAGGTGGAGGAGCTCCCCCATCTGGTGGTGCTTCTTCA1979 
GlyGlyPheProGlyGlyGlyAlaProProSerGlyGlyAlaSerSer 
625630635 
GGCCCCACCATTGAAGAGGTGGATTAAGTCAGTCCAAGAAGAAGGT2025 
GlyProThrIleGluGluValAsp 
640645 
GTAGCTTTGTTCCACAGGGACCCAAAAAGTAACATGGAATAATAAAACTATTTAAATTGG2085 
CACC2089 
(2) INFORMATION FOR SEQ ID NO:14: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 646 amino acids 
(B) TYPE: amino acid 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: 
MetSerLysGlyProAlaValGlyIleAspLeuGlyThrThrTyrSer 
151015 
CysValGlyValPheGlnHisGlyLysValGluIleIleAlaAsnAsp 
202530 
GlnGlyAsnArgThrThrProSerTyrValAlaPheThrAspThrGlu 
354045 
ArgLeuIleGlyAspAlaAlaLysAsnGlnValAlaMetAsnProThr 
505560 
AsnThrValPheAspAlaLysArgLeuIleGlyArgArgPheAspAsp 
65707580 
AlaValValGlnSerAspMetLysHisTrpProPheMetValValAsn 
859095 
AspAlaGlyArgProLysValGlnValGluTyrLysGlyGluThrLys 
100105110 
SerPheTyrProGluGluValSerSerMetValLeuThrLysMetLys 
115120125 
GluIleAlaGluAlaTyrLeuGlyLysThrValThrAsnAlaValVal 
130135140 
ThrValProAlaTyrPheAsnAspSerGlnArgGlnAlaThrLysAsp 
145150155160 
AlaGlyThrIleAlaGlyLeuAsnValLeuArgIleIleAsnGluPro 
165170175 
ThrAlaAlaAlaIleAlaTyrGlyLeuAspLysLysValGlyAlaGlu 
180185190 
ArgAsnValLeuIlePheAspLeuGlyGlyGlyThrPheAspValSer 
195200205 
IleLeuThrIleGluAspGlyIlePheGluValLysSerThrAlaGly 
210215220 
AspThrHisLeuGlyGlyGluAspPheAspAsnArgMetValAsnHis 
225230235240 
PheIleAlaGluPheLysArgLysHisLysLysAspIleSerGluAsn 
245250255 
LysArgAlaValArgArgLeuArgThrAlaCysGluArgAlaLysArg 
260265270 
ThrLeuSerSerSerThrGlnAlaSerIleGluIleAspSerLeuTyr 
275280285 
GluGlyIleAspPheTyrThrSerIleThrArgAlaArgPheGluGlu 
290295300 
LeuAsnAlaAspLeuPheArgGlyThrLeuAspProValGluLysAla 
305310315320 
LeuArgAspAlaLysLeuAspLysSerGlnIleHisAspIleValLeu 
325330335 
ValGlyGlySerThrArgIleProLysIleGlnLysLeuLeuGlnAsp 
340345350 
PhePheAsnGlyLysGluLeuAsnLysSerIleAsnProAspGluAla 
355360365 
ValAlaTyrGlyAlaAlaValGlnAlaAlaIleLeuSerGlyAspLys 
370375380 
SerGluAsnValGlnAspLeuLeuLeuLeuAspValThrProLeuSer 
385390395400 
LeuGlyIleGluThrAlaGlyGlyValMetThrValLeuIleLysArg 
405410415 
AsnThrThrIleProThrLysGlnThrGlnThrLeuThrThrTyrSer 
420425430 
AspAsnGlnProGlyValLeuIleGlnValTyrGluGlyGluArgAla 
435440445 
MetThrLysAspAsnAsnLeuLeuGlyLysPheGluLeuThrGlyIle 
450455460 
ProProAlaProArgGlyValProGlnIleGluValThrPheAspIle 
465470475480 
AspAlaAsnGlyIleLeuAsnValSerAlaValAspLysSerThrGly 
485490495 
LysGluAsnLysIleThrIleThrAsnAspLysGlyArgLeuSerLys 
500505510 
GluAspIleGluArgMetValGlnGluAlaGluLysTyrLysAlaGlu 
515520525 
AspGluLysGlnArgAspLysValSerSerLysAsnSerLeuGluSer 
530535540 
TyrAlaPheAsnMetLysAlaThrValGluAspGluLysLeuGlnGly 
545550555560 
LysIleAsnAspGluAspLysGlnLysIleLeuAspLysCysAsnGlu 
565570575 
IleIleSerTrpLeuAspLysAsnGlnThrAlaGluLysGluGluPhe 
580585590 
GluHisGlnGlnLysGluLeuGluLysValCysAsnProIleIleThr 
595600605 
LysLeuTyrGlnSerAlaGlyGlyMetProGlyGlyMetProGlyGly 
610615620 
PheProGlyGlyGlyAlaProProSerGlyGlyAlaSerSerGlyPro 
625630635640 
ThrIleGluGluValAsp 
645 
(2) INFORMATION FOR SEQ ID NO:15: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 5408 base pairs 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: double 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 1040..1244 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 1569..1772 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 2097..2249 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 2337..2892 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 3104..3306 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 3535..3733 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 3881..4113 
(ix) FEATURE: 
(A) NAME/KEY: exon 
(B) LOCATION: 4445..4629 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:15: 
GAGCTTGAAAGTTCCAGAACGCTGCGGTGAGTGCGTTATCGTGAGGCGGCGCGGTGGGGT60 
GGGTGCGGAAGGGGGCGAGGCGAGGAGTGGAGCCGCGTTGTGATTGTGATTGGGTCTTGT120 
AAGGGCAGCCGGACTCTATTGGCCGGGAACCTAATGCAGGAAGCAGGCGGACCCCTTCTG180 
GAAGGTTCTAAGATAGGGTATAAGAGGCAGGGTGGCGGGCGGAAACCGGTGCTCAGTTGA240 
ACTGCGCTGCAGCTCTTGGTTTTTTGTGGCTTCCTTCGTTATTGGAGCCAGGCCTACACC300 
CCAGGTAAAACCTCTGCTCAAGAGTTGGGTTGTGGGTCTGGGAGCGTGCAGCCTCCACAC360 
AGGCCTGTTGGGCTTGCTGAGGCTTGGGGGTTCTGAGAATCTCGTCGAGGCGAGTGTGCG420 
GCTCCTTCTACCGGCTTAAAGGGCCTCAGTTTTCGGTGGGATGGCAGCGGTATTTGGTTG480 
CAGCCGGCAGACGGAAATGTAGGGAGTGGGCCGCATGGCCCCAGGGGAGGCTGGGAGACG540 
CCCGGCCGCGTGGCGGGGGAGGGTTGCTGCATCGGTTTGCCTGGCGCGCGGGGAAGTGGA600 
GCCAGCGTTTTCTTTCACCCAGTTCCCTGCTTAGTCCAGTCCCACCGTGGTTCTTCAGAG660 
CTGTTCTTGGCGTGCTTCCAGTATGGGGGTACATTCCGGAGTAGTTAAAAGCCCGTTGAC720 
TCCCGGGGGGCACTGGCACCTGGCGAGGGAGGGGAACAGACAGTGCTCAGTTCGGGGTAA780 
GACCACGTGTTGAGCAACGCCCCACGCCGTCTGGGTCGATGGGTCCTTCATCTAGGGCGT840 
GCTGTGCTGCGGTTGGCACGGCAACCTGGACTGCAGCACTAGTTCTGGACCTCGCGCGTG900 
CTTAGACAGGAGGTGATGGGCACTATTACCTCTTGGCAGTGGCCATACGTTTTTCCTGGT960 
TAAGTGTTCTGTTAAGGGATGAGGGAAATATTTTGATTAATTGAATTTTTAAACCAGATT1020 
TTTCTTTTTTTCAGCAACCATGTCCAAGGGACCTGCAGTTGGTATTGATCTTGGCACCAC1080 
CTACTCTTGTGTGGGTGTTTTCCAGCACGGAAAAGTCGAGATAATTGCCAATGATCAGGG1140 
AAACCGAACCACTCCAAGCTATGTCGCCTTTACGGACACTGAACGGTTGATCGGTGATGC1200 
CGCAAAGAATCAAGTTGCAATGAACCCCACCAACACAGTTTTTGGTGAGTTCCTAATTTT1260 
AAATGACAGAACAAATATAAACAGGGCTAGGAAGCACAAAAGTTTATGAAACGTGAGGAG1320 
GGAACTTTTTGATTTTAGAAAAACTGAGCTGAGAGACTTGTTATCAAGTCTGTTATAAAA1380 
CAGGTTGTAGAAACCTTTCAGGCTGAAATCTGGATAACGTAGGAGGTTGAAGTTTGAACC1440 
TTTGCTAGGTATATGGTAGTTGAATTCACCTACCTATGAACTGTTAGGTATTTGAGTAAT1500 
CATGGACTTGAGTTTTATCTGAAGAGCTATGAAATTGAAAGTGTTTTCATTTGACACCTT1560 
TTACAGATGCCAAACGTCTGATTGGACGCAGATTTGATGATGCTGTTGTCCAGTCTGATA1620 
TGAAACATTGGCCCTTTATGGTGGTGAATGATGCTGGCAGGCCCAAGGTCCAAGTAGAAT1680 
ACAAGGGAGAGACCAAAAGCTTCTATCCAGAGGAGGTGTCTTCTATGGTTCTGACAAAGA1740 
TGAAGGAAATTGCAGAAGCCTACCTTGGGAAGGTGAGGTTGGTTTTTCAGTATGGGGTGC1800 
ATTCCGGAGTAGTTAAAAGCCCGATGACTCCCGGGGGCACTGGCACCTGGCGAGGGAGGG1860 
GAACAGATGGGGCTCAGCTCAGGGTTAAGACCACGTGCCCAACAGTGCCCTAGGCTCTCT1920 
AGGTAGATGGGTCTGTCAACACCAGAAACCAGTGAATCTTGACAATTACACAGTAATTTA1980 
CATTTTGGTGGGGGGGGTGCTCCAGCTGTTGTTTCACCAGCATTAATCCATTTGCTGGAG2040 
TTTGCATATATGTAAGTATAATAGTTACCAATCTGTGGTCTTTTCCTTATTCCTAGACTG2100 
TTACCAATGCTGTGGTCACAGTGCCAGCTTACTTTAATGACTCTCAGCGTCAGGCTACCA2160 
AAGATGCTGGAACTATTGCTGGTCTCAATGTACTTAGAATTATTAATGAGCCAACTGCTG2220 
CTGCTATTGCTTACGGCTTAGACAAAAAGGTATGTACCATTTGTGATGCAAGTTCGGATT2280 
ATTTTAAGATTAATTTGATCCATCGTAAATTTAAATGAGATTGTTTTTAACGGCAGGTTG2340 
GAGCAGAAAGAAACGTGCTCATCTTTGACCTGGGAGGTGGCACTTTTGATGTGTCAATCC2400 
TCACTATTGAGGATGGAATCTTTGAGGTCAAGTCTACAGCTGGAGACACCCACTTGGGTG2460 
GAGAAGATTTTGACAACCGAATGGTCAACCATTTTATTGCTGAGTTTAAGCGCAAGCATA2520 
AGAAGGACATCAGTGAGAACAAGAGAGCTGTAAGACGCCTCCGTACTGCTTGTGAACGTG2580 
CTAAGCGTACCCTCTCTTCCAGCACCCAGGCCAGTATTGAGATCGATTCTCTCTATGAAG2640 
GAATCGACTTCTATACCTCCATTACCCGTGCCCGATTTGAAGAACTGAATGCTGACCTGT2700 
TCCGTGGCACCCTGGACCCAGTAGAGAAAGCCCTTCGAGATGCCAAACTAGACAAGTCAC2760 
AGATTCATGATATTGTCCTGGTTGGTGGTTCTACTCGTATCCCCAAGATTCAGAAGCTTC2820 
TCCAAGACTTCTTCAATGGAAAAGAACTGAATAAGAGCATCAACCCTGATGAAGCTGTTG2880 
CTTATGGTGCAGGTAACAATGGTATCTCAATTAACCCTAAAGGCAGGCAGGCCCAAGGTG2940 
ACTCGCTGTGATGAGTGATTGTTAAACATTCGTAGTTTCCACCAAAAGCTTGGCTAATGA3000 
TGGCAACACCTTCCTTGGATGTCTGAGCGAGTGATAGTTAAAACAGGAGCTATGTACTGG3060 
GTTTTCTTTTAACTTCTTTTAACGTTAACTTTTTGTTTGCTAGCTGTCCAGGCAGCCATC3120 
TTGTCTGGAGACAAGTCTGAGAATGTTCAAGATTTGCTGCTCTTGGATGTCACTCCTCTT3180 
TCCCTTGGTATTGAAACTGCTGGTGGAGTCATGACTGTCCTCATCAAGCGTAATACCACC3240 
ATTCCTACCAAGCAGACACAGACCTTCACTACCTATTCTGACAACCAGCCTGGTGTGCTT3300 
ATTCAGGTATGTTTCTGTACTTCTCTTGTTTGGCTTACTGATAACAGATAAAGGGAAGTC3360 
TTGACTGACTCGCTATGATGATGGATTCCAAAACCATTCGTAGTTTCCACCAGAAAGTCT3420 
TATGTTGGCCAGTTCCTTCCTTGGATGTTTGAGCGACCATTCTTCCTTAGCAGGACCCTA3480 
GCACTGTCACAGACCTGGAGTCCATTGTAGTAATTTGTTTTATTTCCTACCAAGGTTTAT3540 
GAAGGCGAGCGTGCCATGACAAAGGATAACAACCTGCTTGGCAAGTTTGAACTCACAGGC3600 
ATACCTCCTGCACCCCGAGGTGTTCCTCAGATTGAAGTCACTTTTGACATTGATGCCAAT3660 
GGTATACTCAATGTCTCTGCTGTGGACAAGAGTACGGGAAAAGAGAACAAGATTACTATC3720 
ACTAATGACAAGGGTAAGGAGGCACTGTCATCTGGTCTTGACAGGGATAATGGTATTTCA3780 
ATTGAGTTACTGGTGAATAAGGGCGTCTAGCTAAGAGAAACTAGAGTTACACATACACAG3840 
GTAATTTAAGGCTTTTACTTAGAGTTAATTTCTTTCCTAGGCCGTTTGAGCAAGGAAGAC3900 
ATTGAACGTATGGTCCAGGAAGCTGAGAAGTACAAAGCTGAAGATGAGAAGCAGAGGGAC3960 
AAGGTGTCATCCAAGAATTCACTTGAGTCCTATGCCTTCAACATGAAAGCAACTGTTGAA4020 
GATGAGAAACTTCAAGGCAAGATTAACGATGAGGACAAACAGAAGATTCTGGACAAGTGT4080 
AATGAAATTATCAACTGGCTTGATAAGAATCAGGTTTGTGTTTTTTTTTTTTTTTTTCCT4140 
CCCCCACGCAATGGAGGGGAAGGGGATGGTAAACCAAGCTTGAGCTGGATTTCAGTGTAG4200 
GGTCACAATGATGAATGGTCCAAAACATTCGCGGTTTCCACCAGAATTCAAGGTGTTGGC4260 
AACTACCTTCCTTGGATGTCTGAGTGACCCAAGATGTTAAGGAAGAATAAGGCCCTATTT4320 
TAATGTTGGTATGGGCCCTCTTGTAAGAGTTTGCTCCAGACTTTTAGTATCAGATTGCGT4380 
CAGGGAGAAAGAAGGGTTATTAACATTAAAAGAACTTGCAGTAATTCCTTTTTCTCTTCC4440 
TCAGACTGCTGAGAAGGAAGAATTTGAACATCAACAGAAAGAGCTGGAGAAAGTTTGCAA4500 
CCCCATCATCACCAAGCTGTACCAGAGTGCAGGAGGCATGCCAGGAGGAATGCCTGGGGG4560 
ATTTCCTGGTGGTGGAGCTCCTCCCTCTGGTGGTGCTTCCTCAGGGCCCACCATTGAAGA4620 
GGTTGATTAAGCCAACCAAGTGTAGATGTAGCATTGTTCCACACATTTAAAACATTTGAA4680 
GGACCTAAATTCGTAGCAAATTCTGTGGCAGTTTTAAAAAGTTAAGCTGCTATAGTAAGT4740 
TACTGGGCATTCTCAATACTTGAATATGGAACATATGCACAGGGGAAGGAAATAACATTG4800 
CACTTTATACACTGTATTGTAAGTGGAAAATGCAATGTCTTAAATAAAACTATTTAAAAT4860 
TGGCACCATACAATTGCTTTGAGTCTTTAAATAATCTCCCAGGCCAGCGGTGGGAGAAGT4920 
AGGCTTAGGTGATTATGTGACTCTTACTTTCTCCTTCCTCTTAAGCTTGAGTTAACAAGG4980 
GCTGGGTGGCAAGTTGCCCTTCAGAGCATGTGGATGGTACATTTTGGAATTCAGAGCTTT5040 
GAGAAGGGGAGCATAAGAAATTGGATCTGGATCAAACTAACCTTAGTCCTTAGGCTGGAG5100 
AGGCAGAAGCTGACTTAATGGTGTTTTCTAAACTTATTCTGTGTGTAAGCCTGCCTAGGA5160 
GCAGAGGCTTTCCTGGAGGGTTGTGCTAGATGAGTAAGAATTTAGATACAGAATCAAATA5220 
ATGGGCAGTGAATATTAAGCTACATGGCAGAGGTATCTGAATGTCAATCCCTTATATGAG5280 
CCACTGCCCTGTGGGCTTCCATTTCTTCTGAGTTAAGATTATTCAGAAGGTCGGGGATTG5340 
GAGCTAAGCTGCCACCTGGTTAATTAAGGTCCCAACAGTGAGTTGTGATAGCCTAGGGGA5400 
GCAGGCTG5408 
(2) INFORMATION FOR SEQ ID NO:16: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 666 amino acids 
(B) TYPE: amino acid 
(C) STRANDEDNESS: single 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: 
GluThrArgArgPheValCysAspGluArgArgAlaGlyGlyMetArg 
151015 
HisLeuLeuLeuAlaLeuLeuLeuLeuGlyGlyAlaArgAlaAspAsp 
202530 
GluGluLysLysGluAspValGlyThrValValGlyIleAspLeuGly 
354045 
ThrThrTyrSerCysValGlyValPheLysAsnGlyArgValGluIle 
505560 
IleAlaAsnAspGlnGlyAsnArgIleThrProSerTyrValAlaPhe 
65707580 
ThrProGluGlyGluArgLeuIleGlyAspAlaAlaLysAsnGlnLeu 
859095 
ThrSerAsnProGluAsnThrValPheAspAlaLysArgLeuIleGly 
100105110 
ArgThrTrpAsnAspProSerValGlnGlnAspIleLysTyrLeuPro 
115120125 
PheLysValValGluLysLysAlaLysProHisIleGlnValAspVal 
130135140 
GlyGlyGlyGlnThrLysThrPheAlaProGluGluIleSerAlaMet 
145150155160 
ValLeuThrLysMetLysGluThrAlaGluAlaTyrLeuGlyLysLys 
165170175 
ValThrHisAlaValValThrValProAlaTyrPheAsnAspAlaGln 
180185190 
ArgGlnAlaThrLysAspAlaGlyThrIleAlaGlyLeuAsnValMet 
195200205 
ArgIleIleAsnGluProThrAlaAlaAlaIleAlaTyrGlyLeuAsp 
210215220 
LysArgGluGlyGluLysAsnIleLeuValPheAspLeuGlyGlyGly 
225230235240 
ThrPheAspValSerLeuLeuThrIleAspAsnGlyValPheGluVal 
245250255 
ValAlaThrAsnGlyAspThrHisLeuGlyGlyGluAspPheAspGln 
260265270 
ArgValMetGluHisPheIleLysLeuTyrLysLysLysThrGlyLys 
275280285 
AspValArgLysAspAsnArgAlaValGlnLysLeuArgArgGluVal 
290295300 
GluLysAlaLysArgAlaLeuSerSerGlnHisGlnAlaArgIleGlu 
305310315320 
IleGluSerPhePheGluGlyGluAspPheSerGluThrLeuThrArg 
325330335 
AlaLysPheGluGluLeuAsnMetAspLeuPheArgSerThrMetLys 
340345350 
ProValGlnLysValLeuGluAspSerAspLeuLysLysSerAspIle 
355360365 
AspGluIleValLeuValGlyGlySerThrArgIleProLysIleGln 
370375380 
GlnLeuValLysGluPhePheAsnGlyLysGluProSerArgGlyIle 
385390395400 
AsnProAspGluAlaValAlaTyrGlyAlaAlaValGlnAlaGlyVal 
405410415 
LeuSerGlyAspGlnAspThrGlyAspLeuValLeuLeuAspValCys 
420425430 
ProLeuThrLeuGlyIleGluThrValGlyGlyValMetThrLysLeu 
435440445 
IleProArgAsnThrValValProThrLysLysSerGlnIlePheSer 
450455460 
ThrAlaSerAspAsnGlnProThrValThrIleLysValTyrGluGly 
465470475480 
GluArgProLeuThrLysAspAsnHisLeuLeuGlyThrPheAspLeu 
485490495 
ThrGlyIleProProAlaProArgGlyValProGlnIleGluValThr 
500505510 
PheGluIleAspValAsnGlyIleLeuArgValThrAlaGluAspLys 
515520525 
GlyThrGlyAsnLysAsnLysIleThrIleThrAsnAspGlnAsnArg 
530535540 
LeuThrProGluGluIleGluArgMetValAsnAspAlaGluLysPhe 
545550555560 
AlaGluGluAspLysLysLeuLysGluArgIleAspAlaArgAsnGlu 
565570575 
LeuGluSerTyrAlaTyrSerLeuLysAsnGlnIleGlyAspLysGlu 
580585590 
LysLeuGlyGlyLysLeuSerSerGluAspLysGluThrIleGluLys 
595600605 
AlaValGluGluLysIleGluTrpLeuGluSerHisGlnAspAlaAsp 
610615620 
IleGluAspPheLysSerLysLysLysGluLeuGluGluValValGln 
625630635640 
ProIleValSerLysLeuTyrGlySerAlaGlyProProProThrGly 
645650655 
GluGluGluAlaAlaGluLysAspGluLeu 
660665 
(2) INFORMATION FOR SEQ ID NO:17: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 2403 base pairs 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: double 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: 
AAGGGGTTGACCGTCCGTCGGCACACCACTTATAATGCGGGGTGCAAGCCCCCCGTCTAA60 
AATTTTTTTTTTTTCCATTTTTGTCGTTATTGTTATTTCCCGTTTTTTGTTTTTTTTGAT120 
TTTTTCGGAGCGACAAACCTTTCGAAACACGTGTCCTGAAAATTATCCTGGGCTGCACGT180 
GATAATATGTTACCCTGTCGGGCGGCGCCTCTTTTTCCCTTTTCTCTCACTAGTCTCTTT240 
TTCCAATTTGCCACCGTGTAGCATTTTGTTGTGCTGTTACAACCACAACAAAACGAAAAA300 
CCCGTATGGACATACATATATATATATATATATATATATATATATTTTGTTACGCGTGCA360 
TTTTCTTGTTGCAAGCAGCATGTCTAATTGGTAATTTTAAAGCTGCCAAGCTCTACATAA420 
AGAAAAACATACATCTATCCCGTTATGAAGTTTTCTGCTGGTGCCGTCCTGTCATGGTCC480 
TCCCTGCTGCTCGCCTCCTCTGTTTTCGCCCAACAAGAGGCTGTGGCCCCTGAAGACTCC540 
GCTGTCGTTAAGTTGGCCACCGACTCTTTCAATGAATACATTCAGTCGCACGACTTGGTG600 
CTTGCGGAGTTTTTTGCTCCATGGTGTGGCCACTGTAAGAACATGGCTCCTGAATACGTT660 
AAAGCCGCCGAGACTTTAGTTGAGAAAAACATTACCTTGGCCCAGATCGACTGTACTGAA720 
AACCAGGATCTGTGTATGGAACACAACATTCCAGGGTTCCCAAGCTTGAAGATTTTCAAA780 
AACAGCGATGTTAACAACTCGATCGATTACGAGGGACCTAGAACTGCCGAGGCCATTGTC840 
CAATTCATGATCAAGCAAAGCCAACCGGCTGTCGCCGTTGTTGCTGATCTACCAGCTTAC900 
CTTGCTAACGAGACTTTTGTCACTCCAGTTATCGTCCAATCCGGTAAGATTGACGCCGAC960 
TTCAACGCCACCTTTTACTCCATGGCCAACAAACACTTCAACGACTACGACTTTGTCTCC1020 
GCTGAAAACGCAGACGATGATTTCAAGCTTTCTATTTACTTGCCCTCCGCCATGGACGAG1080 
CCTGTAGTATACAACGGTAAGAAAGCCGATATCGCTGACGCTGATGTTTTTGAAAAATGG1140 
TTGCAAGTGGAAGCCTTGCCCTACTTTGGTGAAATCGACGGTTCCGTTTTCGCCCAATAC1200 
GTCGAAAGCGGTTTGCCTTTGGGTTACTTGTTCTACAATGACGAGGAAGAATTGGAAGAT1260 
TACAAGCCTCTCTTTACCGAGTTGGCCAAAAAGAACAGAGGTCTAATGAACTTTGTTAGC1320 
ATCGATGCCAGAAAATTCGGCAGACACGCCGGCAACTTGAACATGAAGGAACAATTCCCT1380 
CTATTTGCCATCCACGACATGACTGAAGACTTGAAGTACGGTTTGCCTCAACTCTCTGAA1440 
GAGGCGTTTGACGAATTGAGCGACAAGATCGTGTTGGAGTCCAAGGCTATTGAATCTTTG1500 
GTTAAGGACTTCTTGAAAGGTGATGCCTCCCCAATCGTGAAGTCCCAAGAGATCTTCGAG1560 
AACCAAGATTCCTCTGTCTTCCAATTGGTCGGTAAGAACCATGACGAAATCGTCAACGAC1620 
CCAAAGAAGGACGTTCTTGTTTTGTACTATGCCCCATGGTGTGGTCACTGTAAGAGATTG1680 
GCCCCAACTTACCAAGAACTAGCTGATACCTACGCCAACGCCACAACCGACGTTTTGATT1740 
GCTAAACTAGACCACACTGAAAACGATGTCAGAGGCGTCGTAATTGAAGGTTACCCAACA1800 
ATCGTCTTATACCCAGGTGGTAAGAAGTCCGAATCTGTTGTGTACCAAGGTTCAAGATCC1860 
TTGGACTCTTTATTCGACTTCATCAAGGAAAACGGTCACTTCGACGTCGACGGTAAGGCC1920 
TTGTACGAAGAAGCCCAGGAAAAAGCTGCTGAGGAAGCCGATGCTGACGCTGAATTGGCT1980 
GACGAAGAAGATGCCATTCACGATGAATTGTAATTCTGATCACTTTGGTTTTTCATTAAA2040 
TAGAGATATATAAGAAATTTTCTAGGAAGTTTTTTTAAAAAAAATCATAAAAAGATAAAC2100 
GTTAAAATTCAAACACAATAGTCGTTCGCTATATTCGTCACACTGCACGAACGCCTTAGG2160 
GAAAGAGAAAATTGACCACGTAGTAATAATAAGTGCATGGCATCGTCTTTTACTTAAATG2220 
TGGACACTTGCTTTACTGCTTAGGAAACTACTTATCTCATCCTCCTCCATTCCCCTCCCT2280 
TTTCCAATTACCGTAATAAAAGATGGCTGTATTTACTCCTCCATCAGGTAATAGCAATTC2340 
CGACCATACTCACACACAAGATGACCACGACAAAGATGATATGATATCAAGAAATTCTAT2400 
ACA2403 
(2) INFORMATION FOR SEQ ID NO:18: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 504 amino acids 
(B) TYPE: amino acid 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: protein 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: 
MetLysPheSerAlaGlyAlaValLeuSerTrpSerSerLeuLeuLeu 
151015 
AlaSerSerValPheAlaGlnGlnGluAlaValAlaProGluAspSer 
202530 
AlaValValLysLeuAlaThrAspSerPheAsnGluTyrIleGlnSer 
354045 
HisAspLeuValLysAlaAlaGluThrLeuValGluLysAsnIleThr 
505560 
LeuAlaGlnIleAspCysThrGluAsnGlnAspLeuCysMetGluHis 
65707580 
AsnIleProGlyPheProSerLeuLysIlePheLysAsnSerAspVal 
859095 
AsnAsnSerIleAspTyrGluGlyProArgThrAlaGluAlaIleVal 
100105110 
GlnProMetIleLysGlnSerGlnProAlaValAlaValValAlaVal 
115120125 
ValAlaAspLeuProAlaTyrLeuAlaAsnGluThrPheValThrPro 
130135140 
ValIleValGlnSerGlyLysIleAspAlaAspPheAsnAlaThrPhe 
145150155160 
TyrSerMetAlaAsnLysHisPheAsnAspTyrAspPheValSerAla 
165170175 
GluAsnAlaAspAspAspPheLysLeuSerIleTyrLeuProSerAla 
180185190 
MetAspGluProValValTyrAsnGlyLysLysAlaAspIleAlaAsp 
195200205 
AlaAspValPheGluLysTrpLeuGlnValGluAlaLeuProTyrPhe 
210215220 
GlyGluIleAspGlySerValPheAlaGlnTyrValGluSerGlyLeu 
225230235240 
ProLeuGlyTyrLeuPheTyrAsnAspGluGluGluLeuGluGluTyr 
245250255 
LysProLeuPheThrGluLeuAlaLysLysAsnArgGlyLeuMetAsn 
260265270 
PheValSerIleAspAlaArgLysPheGlyArgHisAlaGlyAsnLeu 
275280285 
AsnMetLysGluGlnPheProLeuPheAlaIleHisAspMetThrGlu 
290295300 
AspLeuLysTyrGlyLeuProGlnLeuSerGluGluAlaPheAspGlu 
305310315320 
LeuSerAspLysIleValLeuGluSerLysAlaIleGluSerLeuVal 
325330335 
LysAspPheLeuLysGlyAspAlaSerProIleValLysSerGlnGlu 
340345350 
IlePheGluAsnGlnAspSerSerValPheGlnLeuValGlyLysAsn 
355360365 
HisAspGluIleValAsnAspProLysLysAspValLeuValLeuTyr 
370375380 
AlaProTrpCysGlyHisCysLysArgLeuAlaProThrTyrGlnGlu 
385390395400 
LeuAlaAspThrTyrAlaAsnAlaThrSerAspValLeuIleAlaLys 
405410415 
LeuAspHisThrGluAsnAspValArgGlyValValIleGluGlyTyr 
420425430 
ProThrIleValLeuTyrProGlyGlyLysLysSerGluSerValVal 
435440445 
TyrGlnGlySerArgSerLeuAspSerLeuPheAspProIleLysGlu 
450455460 
AsnGlyHisPheAspValAspGlyLysAlaLeuTyrGluGluAlaGln 
465470475480 
GluLysAlaAlaGluGluAlaAspAlaAspAlaGluLeuAlaAspGlu 
485490495 
GluAspAlaIleHisAspGluLeu 
500 
(2) INFORMATION FOR SEQ ID NO:19: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 2473 base pairs 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: double 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: 
CCCCGGCGCCAACCTAGCTGCCCCGCCCGCTGCCGACGTCCGACATGCTGAGCCGTGCTT60 
TGCTGTGCCTGGCCCTGGCCTGGGCGGCTAGGGTGGGCGCCGACGCTCTGGAGGAGGAGG120 
ACAACGTCTCGGTGCTGAAGAAGAGCAACTTCGCAGAGCCGGCGGCGCACAACTACCTGC180 
TGGTGGAGTTCTATGCCCCATGGTGTGGCCACTGCAAAGCATCGGCCCCAGAGTATGCCA240 
AAGCTGCTGCAAAACTGAAGGCAGAAGGACTCGAGATCCGACTAGCAAAGGTGGACGCCA300 
CAGAAGAGTCTGACCTGGCCCAGCAGTATGGTGTCCGTGGCTACCCCACAATCAAGTTCT360 
TCAAGAATGGAGACACAGCCTCCCCAAAGGAATATACAGCTGGCACGGAAGCTGACGACA420 
TTGTGAACTGGCTGAAGAAACGCACAGGCCCAGCAGCCACAACCCTGTCTGACACTGCAG480 
CTGCAGAGTCCTTGCTGGACTCAAGCGAAGTGACGGCTATCGGCTTCTTCAAGGACGCAG540 
GGTCAGACTCCGCCAAGCAGTTCTTGCTGGCAGCAGAGGCTGCTGATGACATACCTTTTG600 
GAATCACTTCCAATTGCGTGTTTTCCAAGTACCAGCTGGACAACGATGGGGTGGTCCTCT660 
TTAAGAAGTTTGATGAAGGCCGCAACAATTTTGAATGGTGAGATCACCAAGGAGAAGCTA720 
TTAGACTTCATCAAGCACAACCAGCTGCCTTTGGTCATCGAGTTCACTGAACAGACAGCT780 
CCAAAGATTTTCGGAGGTGAAATCAAGACACATATTCTGCTGTTCCTGCCCAAGAGTGTG840 
TCTGACTACGATGGCAAATTGAGCAACTTTAAGAAAGCGGCCGAGGGCTTTAAGGGCAAG900 
ATCCTGTTCATCTTCATCGATAGTGACCACACTGACAACCAGCGCATACTTGAGTTCTTT960 
GGCCTGAAGAAGGAGGAATGTCCAGCTGTGCGGCTTATTACCCTGGAGGAAGAGATGACC1020 
AAGTACAAACCGGAGTCAGACGAGCTGACAGCTGAGAAGATCACACAATTTTGCCACCAC1080 
TTCCTGGAGGGCAAGATCAAGCCCCACCTGATGAGCCAGGAACTGCCTGAAGACTGGGAC1140 
AAGCAGCCAGTGAAAGTGCTAGTTGGGAAAAACTTTGAGGAGGTTGCTTTTGATGAGAAA1200 
AAGAACGTGTTTGTTGAATTCTATGCTCCCTGGTGTGGTCACTGCAAGCAGCTAGCCCCG1260 
ATTTGGGATAAACTGGGAGAGACATACAAAGACCATGAGAATATCGTCATCGCTAAGATG1320 
GACTCAACAGCCAATGAGGTGGAAGCTGTGAAGCTGCACACCTTTCCCACACTCAAGTTC1380 
TTCCCAGCAAGTGCAGACAGAACGGTCATTGATTACAACGGTCAGCGGACACTAGATGGT1440 
TTTAAGAAATTCTTGGAGAGCGGTGGCCAGGATGGAGCGGGGGACAATGACGACCTCGAC1500 
CTAGAAGAAGCTTTAGAGCCAGATATGGAAGAAGACGACGATCAGAAAGCCGTGAAGGAT1560 
GAACTGTAGTCGAGAAGCCAGATCTGGCGCCCTGAACCCAAAACCTCGGTGGGCCATGTC1620 
CCAGCAGCCCACATCTCCGGAGCCTGAGCCTCACCCCAGGAGGGAGCGCCATCAGAACCC1680 
AGGGAATCTTTCTGAAGCCACACTCATCTGACACACGTACACTTAAACCTGTCTCTTCTT1740 
TTTTTGCTTTTCAATTTTGGAAAGGGATCTCTGTCCAGGCCAGCCCATCTTGAAGGGCTA1800 
CGTTTTGTTTTAATTGGTGGTGTACTTTTTTGTACGTGGATTTTGTCCCAAGTGCTTGCT1860 
ACCATATTTGGGGATTTCACACTGGTAATGTCTTTCCTGTTAGAGAGGTTTATGCTATCA1920 
CTTCAGATTTCGTCTGTGAGATCTTTCATCTTCCTGACATGTTCTCATGTCGAGGTACTT1980 
GTTCCACCACGCAGATTCCCCTGAGACCCCTTCCTGCCCTGCGCAGGAGGCGATCGTTCT2040 
GGGTCGTATGCTCTCTCTCTCTCCACCTTGTACTAGTGTTGCCATGACAGCTAGGCTTTT2100 
GTAGTTTGCATTTAACCTGGGGATTTCTGCATCCTGTCAGAGGCTGGGTCCCCACGTGTG2160 
GAAAAGAGACAGTGGTGGCTTGCTGCCAGGCACAGGCCAGGCCTGGACAGCTCTCACTCT2220 
TCTTAAGCCAGAACTACCGACCAGCCGGCCGGCTGTCCGCACATTACTCTGGCTCCTGGA2280 
TCCTCTTCCAGCATGGCATGTGGCCTGTGTGAGGCAGAACCGGGACCCTTGATTCCCAGA2340 
CTGGGAGTCAGCTAAGGACACTGGCGCTGAATGAAATGCCCATTCTCAAGGTCTATTTCT2400 
AAACCATAATGTTGGAATTGAACACATTGGCTAAATAAAGTTGAAATTTTACTACCATAA2460 
AAAAAAAAAAAAA2473 
(2) INFORMATION FOR SEQ ID NO:20: 
(i) SEQUENCE CHARACTERISTICS: 
(A) LENGTH: 510 amino acids 
(B) TYPE: amino acid 
(D) TOPOLOGY: linear 
(ii) MOLECULE TYPE: DNA (genomic) 
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: 
MetLeuSerArgAlaLeuLeuCysLeuAlaLeuAlaTrpAlaAlaArg 
151015 
ValGlyAlaAspAlaLeuGluGluGluAspAsnValLeuValLeuLys 
202530 
LysSerAsnPheAlaGluProAlaAlaHisAsnTyrLeuLeuValGlu 
354045 
PheTyrAlaProTrpCysGlyHisCysLysAlaLeuAlaProGluTyr 
505560 
AlaLysAlaAlaAlaLysLeuLysAlaGluGlySerGluIleArgLeu 
65707580 
AlaLysValAspAlaThrGluGluSerAspLeuAlaGlnGlnTyrGly 
859095 
ValArgGlyTyrProThrIleLysPhePheLysAsnGlyAspThrAla 
100105110 
SerProLysGluTyrThrAlaGlyArgGluAlaAspAspIleValAsn 
115120125 
TrpLeuLysLysArgThrGlyProAlaAlaThrThrLeuSerAspThr 
130135140 
AlaAlaAlaGluSerLeuValAspSerSerGluValThrValIleGly 
145150155160 
PhePheLysAspAlaGlySerAspSerAlaLysGlnPheLeuLeuAla 
165170175 
AlaGluAlaValAspAspIleProPheGlyIleThrSerAsnSerAsp 
180185190 
ValPheSerLysTyrGlnLeuAspLysAspGlyValValLeuPheLys 
195200205 
LysPheAspGluGlyArgAsnAsnPheGluGlyGluIleThrLysGlu 
210215220 
LysLeuLeuAspPheIleLysHisAsnGlnLeuProLeuValIleGlu 
225230235240 
PheThrGluGlnThrAlaProLysIlePheGlyGlyGluIleLysThr 
245250255 
HisIleLeuLeuPheLeuProLysSerValSerAspTyrAspGlyLys 
260265270 
LeuSerAsnPheLysLysAlaAlaGluGlyPheLysGlyLysIleLeu 
275280285 
PheIlePheIleAspSerAspHisThrAspAsnGlnArgIleLeuGlu 
290295300 
PhePheGlyLeuLysLysGluGluCysProAlaValArgLeuIleThr 
305310315320 
LeuGluGluGluMetThrLysTyrLysProGluSerAspGluLeuThr 
325330335 
AlaGluLysIleThrGlnPheCysHisHisPheLeuGluGlyLysIle 
340345350 
LysProHisLeuMetSerGlnIleGluLeuProGluAspTrpAspLys 
355360365 
GlnProValLysValLeuValGlyLysAsnPheGluGluValAlaPro 
370375380 
AspGluLysLysAsnValPheValGluPheTyrAlaProTrpCysGly 
385390395400 
HisCysLysGlnLeuAlaProIleTrpAspLysLeuGlyGluThrTyr 
405410415 
LysAspHisAspGluAsnIleValIleAlaLysMetAspSerThrAla 
420425430 
AsnGluValGluAlaValLysValHisSerPheProThrLeuLysPhe 
435440445 
PheProAlaSerAlaAspArgThrValIleAspTyrAsnGlyGluArg 
450455460 
ThrLeuAspGlyPheLysLysPheLeuGluSerGlyGlyGlnAspGly 
465470475480 
AlaGlyAspAsnAspAspLeuAspLeuGluGluAlaLeuGluProAsp 
485490495 
MetGluGluAspAspAspGlnLysAlaValLysAspGluLeu 
500505510 
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