DNA compounds comprising sequences encoding mannuronan C-5-epimerase

Abstract

DNA compounds encompassing sequences coding for enzymes having mannuronan C-5-epimerase activity are disclosed and a process for the preparation of such enzymes. The genetic sequences and enzymes prepared may be used in the production of alginates having a definite G/M ratio and block structure. Alginates having a definite G/M ratio may also be produced by selective inactivation of the genetic sequences.

Description

The present invention concerns DNA compounds encompassing sequences coding for enzymes having mannuronan C-5-epimerase activity, a process for the preparation of such enzymes, the use of said genetic sequences in production of alginates having a definite G/M ratio and block structure, and the production of alginates having a definite G/M ratio by inactivating said genetic sequences.

Throughout this application, reference is made to publications from the scientific and patent literature. Publications so cited are hereby incorporated in their entirety by reference.

In this application the term gene is used to indicate a genetic sequence which encodes a protein, independent of whether the protein encoded by this genetic sequence is expressed or not in the natural host organism under those conditions.

Alginates are a family of polysaccharides, which are synthesized in brown algae as well as in bacteria, such as Azotobacter vinelandii and Azotobacter chroococcum. Alginates are also synthesized by some strains of Pseudomonas sp.

Chemically, alginates are unbranched binary copolymers of 1-4 linked .beta.-D-mannuronic acid, termed M, and its C-5 epimer .alpha.-L-guluronic acid, termed G.

Alginates derived from seaweeds and Azotobacter are generally true block copolymers where the monomers are arranged in homopolymeric stretches of M, termed M blocks, and homopolymeric stretches of G, termed G blocks, interspaced with regions containing both monomers, normally termed alternating blocks or MG blocks. The composition and sequential structure of alginates vary widely depending on the source. Alginates produced by Pseudomonas, however, do not have any G blocks.

Several functional properties such as the capacity to form gels and the binding of water depend on the M/G ratio and on the length of the various blocks. A relatively high content of G blocks, for instance gives good gelling properties, due to ionic cross linking of chains which takes place when Ca.sup.2+ -ions are added to an alginate solution. The composition and block structure also influence on the immunological properties of alginates. �Otterlei et al, J. of Immunotherapy 10, 286-291, (1991)! have shown that alginates with a high content of mannuronic acid blocks are very potent nontoxic immunostimulants.

At present industrial production of alginates rely exclusively on algal sources. The range in composition is however limited as the highest content of guluronic acid to be found is 75% and the lowest 25%. Furthermore there are no suitable sources for alginate with a G content in the range of 42-54%. In the field of biotechnology or biomedicine, polymers with extreme compositions, such as a high G for immobilization of cells, �Martinsen A., Skj.ang.k-Br.ae butted.k G. and Smidsr.o slashed.d O., Biotechnol. Bioeng. 33, 79-86, (1989)! and a high M (90-100%) as immunostimulants �Otterlei et al, J. of Immunotherapy 10, 286-291, (1991)! are of major interest.

The key enzyme responsible for generation of the G blocks is called mannuronan C-5-epimerase. It was previously thought that only one enzyme having a certain amino acid sequence would exhibit this activity. It has now surprisingly been found that there exist at least five genes encoding enzymes having this activity. Some of these enzymes differ in molecular weight and amino acid sequence. The genes were found adjacent to each other in the bacterium Azotobacter vinelandii. It has also been found that the amino acid sequence of the enzyme affects the activity of the enzyme, not only in terms of potency but also in the type of alginate formed, for example, altering the content of guluronic acid and the single/block G content of the alginate.

In �Larsen, B. and Haug, A., Carbohydr. Res. 17, (1971), 287-296 and 297-308! the isolation of mannuronan C-5-epimerase from liquid cultures of Azotobacter vinelandii is reported. In the following, this epimerase will be termed mannuronan C-5-epimerase (2), and the DNA sequence encoding for it will correspondingly be denominated E2.

In �Skj.ang.k-Br.ae butted.k, G and Larsen, B, Carbohydr. Res. 103:133-136, (1982)!, the purification of mannuronan C-5-epimerase (2) by affinity chromatography on alginate sepharose is disclosed. In a separate paper �Skj.ang.k-Br.ae butted.k, G and Larsen, B., Carbohydrate Research, 139, (1985) 273-283! the characterization of this enzyme is disclosed. Further, the activity of the enzyme is described as an ability to epimerize both bacterial and algal alginate having a wide range in monomer composition and sequence of units.

From PCT/WO 86/03781 and Japanese Patent Application J63233797 it is known to produce alginic acid and/or alginate having a high content of guluronic acid by action of the enzyme (E2) on an alginic acid or alginate, whereby the G content increases.

In �Chitnis, C. E. and Ohman, D. E., J. Bacteriol., 172, p2894-2900, (1990)! the gene sequences involved in the introduction of guluronic acid into exopolysaccharides from Pseudomonas aeruginosa have been reported. However, the nature of the enzyme responsible for this process has not been identified. Since this genus of bacteria is unable to produce alginate containing G blocks �Skj.ang.k-Br.ae butted.k, G., Larsen, B. and Grasdalen, H. Carbohydr. Res. 54 (1986) 169-174! it is believed that the epimerization system in alginate producing Pseudomonas is fundamentally different from the epimerase in brown algae and in Azotobacter vinelandii. It seems likely that the Pseudomonas enzyme is a monomer epimerase acting at the sugar nucleotide level, and as such is unable to introduce G-blocks into already polymerized alginates.

Production of mannuronan C-5-epimerase from Azotobacter vinelandii culture is difficult due to a very low yield. It is also a major obstacle that the enzyme is secreted together with copious amounts of highly viscous alginate which hampers the purification of the enzyme. Although alginates are secreted by some bacteria, an industrial production based on these microorganisms has not been successful. The main reasons are due to the difficulties in controlling the composition and molecular size of the exopolysaccharides. The content of guluronic acid blocks in the alginate from Azotobacter vinelandii tends to be too low for making a polymer with good gelling properties.

Alginates with a high M content having immunogenic properties as reported above, are produced by Pseudomonas aeruginosa, but this organism is unattractive from a production point of view, as it is unstable in the production of the polymer. Further, the organism is known to be a secondary pathogen in patients suffering from cystic fibrosis.

Thus, in order to produce medical grade alginates with a defined monomer composition and sequential structure there is a need for improved methods for controlling the biosynthesis of alginate, through controlling the key enzyme, the mannuronan C-5-epimerase.

The present invention is directed to cloned DNA fragments encoding mannuronan C-5-epimerase. The invention is encompassed by vectors which contain DNA fragments encoding mannuronan C-5-epimerase linked to DNA elements which direct the expression of mannuronan C-5-epimerase from the cloned DNA encoding the protein. The invention also provides for microorganisms which express the mannuronan C-5-epimerase protein from the cloned DNA as a source of the purified protein and also as a source of alginates of altered composition. Strains in which the expression level of the mannuronan C-5-epimerase gene is altered or in which one, several or all of the mannuronan C-5-epimerase genes have been inactivated are also within the scope of the present invention. The invention further encompasses methods for producing alginates either very efficiently, or having altered composition, or both, by culturing microorganisms having altered levels of expression of a mannuronan C-5-epimerase gene.

The invention further features selection of epimerase to achieve a desired level of guluronic acid, and alter the single/block G characteristics of the enzyme. In a further embodiment, the invention features the production of synthetic proteins and DNA encoding such proteins which have mannuronan C-5-epimerase activity.

BRIEF DESCRIPTION OF FIGURES

FIG. 1 shows the amino acid sequence SEQ ID NO:6 of the N-terminal end of the 122 kd protein, and the nucleotide sequence of the corresponding oligonucleotide SEQ ID NO:16. The DNA probe was synthesized as a mixture (in equal ratios) of the 64 possible combinations that could be deduced from the first seven amino acids in the sequence of the 122 kd protein. N indicates that all four bases were used at this position.

FIG. 2 is a restriction endonuclase map of the combined inserts in plasmids pHE14, pHE16, pBD1, pHE18 and PML1.

The numbers at the bottom line indicate the molecular sizes in bp. The arrow indicates the localization and orientation of the sequence homologous to the synthetic oligonucleotide used for screening the library. The five genes (open reading frames) found by sequencing are marked by boxes and denoted E4, E1, E2, E3 and E5. E1 corresponds to Epimerase I.

FIG. 3 shows Mannuronan C-5-epimerase (1) activity of a portion of the E1 encoded protein as a function of cell growth. *: OD.sub.600 of cell culture. o: Epimerase activity given as dpm/ml of cell culture. The strain used in this experiment was DH5.alpha.(pHE5), and the extracts were incubated with the substrate for 23 hours.

FIG. 4 shows the kinetics of .sup.3 H release. The enzyme activity was assayed by using an extract prepared from IPTG-induced JM105 cells containing pHE5 (see legend to Table 3).

FIG. 5 shows the homologies between and within the different genes. Boxes with the same letter are homologous to each other. Gaps are introduced to optimize the alignment. E1-E4 are defined as appears from FIGS. 2 and 6.

FIGS. 6A-6E (SEQ ID NO:7) shows the alignment of the DNA sequences of the A blocks from E4, E1, E2 and E3 SEQ ID NO:7=Con, SEQ ID NO:17=E4A, SEQ ID NO:18=E1A1, SEQ ID NO:19=E1A2, SEQ ID NO:20=E2A, SEQ ID NO:21=E3A1, SEQ ID NO:22=E3A2.

FIG. 7A-7B (SEQ ID NO:8) shows the alignment of the deduced amino acid sequences of the A blocks from E4, E1, E2 and E3 SEQ ID NO:8=Con, SEQ ID NO:23=E4A, SEQ ID NO:24=E1A1, SEQ ID NO:25=E1A2, SEQ ID NO:26=E2A, SEQ ID NO:27=E3A1, SEQ ID NO:28=E3A2.

FIGS. 8A-8D (SEQ ID NO:9) shows the alignment of the DNA sequences of the R blocks from E4, E1, E2 and E3 SEQ ID NO:9=Con, SEQ ID NO:29=E4R1, SEQ ID NO:30=E1R1, SEQ ID NO:31=E1R2, SEQ ID NO:32=E1R3, SEQ ID NO:33=E1R4, SEQ ID NO:34=E2R1, SEQ ID NO:35=E2R2, SEQ ID NO:36=E2R3, SEQ ID NO:37=E2R4, SEQ ID NO:38=E3R1, SEQ ID NO:39=E3R2, SEQ ID NO:40=E3R3.

FIG. 9A-9B (SEQ ID NO:10) shows the alignment of the deduced amino acid sequences from the R blocks of E4, E1, E2 and E3 SEQ ID NO:10=Con, SEQ ID NO:41=E4R1, SEQ ID NO:42=E1R1, SEQ ID NO:43=E1R2, SEQ ID NO:44=E1R3, SEQ ID NO:45=E1R4, SEQ ID NO:46=E2R1, SEQ ID NO:47=E2R2, SEQ ID NO:48=E2R3, SEQ ID NO:49=E2R4, SEQ ID NO:50=E3R1, SEQ ID NO:51=E3R2, SEQ ID NO:52=E3R3.

FIG. 10A-10C shows .sup.1 H-NMR spectra of alginate epimerized by extracts from A: DH5.alpha.(pHE8) (truncated epimerase 1); B: JM109(pBD9); C: no extract. The peak to the left gives the signal from G-1; the peak in the centre gives the combined signal from GM-5 and M-1 and the peak to the right gives the signal from GG-5.

FIGS. 11A-11F (SEQ ID NO:1) shows the nucleotide sequence and corresponding amino acid sequence of E2.

Now according to the present invention genetic sequences have been found which encode enzymes having mannuronan C-5-epimerase activity (MG copolymers), and thus the first aspect of the invention is pure isolated DNA comprising nucleotide sequences encoding mannuronan C-5-epimerase activity (GGhorropolymers).

The sequence of amino acids proximal to the amino terminus of purified mannuronan C-5-epimerase protein was determined �G. Skj.ang.k Br.ae butted.k et al., Carbohydr. Res. 103:133-136 (1982)!. This data was used to derive a sequence for an oligonucleotide probe which was used to screen a gene library of Azotobacter vinelandii DNA. One result of this screening experiment was the surprising discovery of a second gene and thereafter three further genes including at least one genetic block A were found. Thus, altogether there appear to be at least five different genes encoding proteins having mannuronan C-5-epimerase activity. Accordingly, it is a second object of the present invention to provide for alternative DNA sequences encoding mannuronan C-5-epimerase.

Three different blocks of genetic sequences, designated A, R and S, are found in the genes. These genetic blocks are most commonly found in combinations wherein the A appears one time or two times, the R block appears from 0 to at least 5 times and the S block appears from 0-1 time.

There is a high degree of consensus in the nucleotide sequences of each block for the different genes (1-5). Accordingly, it is a third object of the present invention to provide for DNA sequences encoding mannuronan C-5-epimerase and comprising the DNA blocks A and/or S and/or R, wherein A may appear more than once and R if present may appear singly or in repeats of up to at least 5 or 6 times.

The consecutive order of the three blocks if all three blocks are present, is preferably A, R and S. However, it has been shown that a reversed consecutive order, wherein for instance R appears before A also gives a gene encoding a mannuronan C-5-epimerase. Thus, the invention further encompasses genetic sequences having any order and any number of the blocks A, R and S.

Another aspect of the present invention concerns the use of said genetic sequences for the preparation of the mannuronan C-5-epimerase in recombinant host cells. It is especially preferred to insert the gene into hosts such as bacteria, for instance Escherichia coli or Bacillus subtilis or in yeast. The cloning and expression of the genetic sequence as described above in E. coli is described in the Examples.

The present invention also encompasses recombinant expression plasmids that can be used to produce the mannuronan C-5-epimerase proteins in a host microorganism. Such expression plasmids are made by inserting a DNA fragment encoding mannuronan C-5-epimerase into a vector which contains appropriate expression elements, such as (but not limited to) a promoter, ribosome binding site, translational initiation site and transcription terminator. The expression plasmids can be adapted for transformation into many different commonly used host organisms in which it might be desired to produce the mannuronan C-5-epimerase.

The techniques for insertion of foreign genes into commonly employed hosts are known in the art, as described for instance in �METHODS IN ENZYMOLOGY, Vol. 185, Gene Expression Technology, Ed. D. V. Goeddel, Academic Press, Inc. (1990)!. Further by choice of a broad host range vector and a suitable promoter as known in the art, and described for instance in �J. L. Ramos et al, FEBS Letters, Vol. 226, 2, 241-246! it will be possible to insert and express the mannuronan C-5-epimerase genetic sequences in many different hosts.

This will make possible the production of large quantities of one or all of the pure enzymes having this activity, while avoiding the problems of separating the enzymes from the alginate.

By inserting a high copy-number vector comprising the genetic sequences encoding the epimerase into a natural alginate producing bacterium such as Azotobacter vinelandii an enhanced production of the enzymes would be possible.

Over expression of the epimerases in a natural host could also be achieved by using a promoter which drives high-level expression of the enzymes. By blocking other genetic sequences coding for the alginate production, the production of pure enzymes may be achieved.

Yet another aspect of the invention is the selective inactivation of the mannuronan C-5-epimerase genes in the natural host organism so as to provide for bacterial production of alginates having a low content of G blocks or even a pure poly-M alginate. This is accomplished by inserting nucleotides into one, several or all of the mannuronan C-5-epimerase genes in the natural host organism Azotobacter. It is especially preferred to insert a DNA fragment encoding a selectable marker gene, preferably a gene conferring antibiotic resistance. Insertion of a selectable marker allows selection of those bacteria in which the insertion has been successfully accomplished. By choosing different selectable markers, for example providing resistance to different antibiotics, it is possible to select recombinants that have incorporated inserted sequences into some or all of the mannuronan C-5-epimerase genes. Thus, selective production of bacterial strains in which one of the mannuronan C-5-epimerase genes, several or all of them have been inactivated is possible.

A second method of inactivating all of the epimerase genes is to transform a cell of the natural host strain, Azotobacter with a vector which expresses an antisense RNA which specifically binds to mRNA transcribed from these genes. Use of promoters of varying strength to drive expression of the antisense RNA in the creation of the vectors used to transform the cells allows production of strains having varying ratios of G-blocks to M-blocks in the alginate produced. Use of inducible promoters to drive expression of the antisense RNA allows the creation of strains which can produce alginates of variable composition, depending on culture conditions. Clearly, if the recombinant host organism is the natural host, Azotobacter, it is possible to enhance production of one epimerase gene while leaving expression of the others at their normal level, thus producing a strain which makes an alginate having an altered ratio of G blocks to M blocks. A strain which makes alginate having 0-25% M blocks is preferred.

Alternatively, all but one of the epimerase genes can be inactivated, as described above, and the expression of the remaining epimerase gene can be controlled by a regulated promoter. A strain carrying such a complement of epimerase genes would thus produce alginates having a high content of G-blocks, especially from 75-98%. Another means for making a strain for producing alginates having a high G-block content is to inactivate all but one of the mannuronan C-5-epimerase genes by insertion and control the remaining gene by antisense RNA, using an inducible promoter to regulate transcription of the antisense RNA gene. A still further means for making a strain for producing alginates having a high G-block content is by inactivating all naturally occurring genes and introducing a regulated gene through a vector.

Thus the present invention also includes a process for the construction of a recombinant host cell capable of expressing mannuronan C-5-epimerase activity by transforming said host cell with a recombinant DNA expression vector that comprises: (a) a promoter and translational activating sequence that function in said host cell; and (b) a DNA sequence encoding mannuronan C-5-epimerase comprising at least a DNA block A and/or a DNA block S and/or a DNA block R, positioned for expression from said promoter and translational activity sequence.

Also the present invention encompasses a process for the bacterial production of pure poly-M alginate or tailored alginates having a lower G block content, preferably in the range from 0-25%, by blocking the DNA sequences encoding the enzymes in a natural host by insertion of a foreign genetic sequence into one, several or all genetic sequences encoding mannuronan C-5-epimerase.

Other methods for achieving the same end will be known for persons skilled in the art and are hereby included into the scope of the present invention.

A further aspect of the invention are the novel enzymes having mannuronan C-5-epimerase activity. The amino acid sequences and their degree of homology will appear from FIGS. 6-9 (SEQ ID NOS:7-10 and SEQ ID NO:1).

Also as known by a person skilled in the art, variations in the nucleotide sequence which nevertheless encode proteins having the same activity as the wild-type mannuronan C-5-epimerase are encompassed within this invention.

Variations within the amino acid sequence may also encompass deletions, substitutions and additions which do not substantially change the biological activity.

Also, it is possible to make a synthetic DNA sequence encoding a mannuronan C-5-epimerase by techniques well known in the art. See for instance, �Itakura et al., Science 198:1056 (1977)! and �Crea et al. (Proc. Natl. Acad. Sci. USA 75:5765 (1978)! and also U.S. Pat. Nos. 4,800,159 and 4,683,202 and also published European patent application EP-A-0258017. Synthetic enzymes may be made by incorporating different combinations of the A, R and S elements, to maintain epimerase activity. The resultant alginate composition can be varied by enzyme selection.

Materials and General Methods

Bacterial strains, plasmids, and phage. Strains, plasmids, and phages are listed in Table 1.

The bacterial strain of A. vinelandii used in these experiments, is freely available from Bj.o slashed.rn Larsen, Inst. of Biotechnology, Lab. for Marine Biochemistry, 7034 Trondheim--NTH, Norway or Svein Valla, Unigen, Center for Molecular Biology, University of Trondheim, 7005 Trondheim, Norway and has been deposited Oct. 4, 1993 at the Belgian Coordinated Collections of Microorganisms (BCCM) at the Laboratorium voor Microbiologie (LMG) at Universiteit Gent (RUG) under the accession number LMG P-14235,

K. L. Ledeganckstraat 35, B-9000 Gent. Other strains of A.vinelandii mentioned in Example 9 have the following ATCC numbers: ATCC 478, ATCC 12837 and ATCC 12518. Plasmids/strains DH5.alpha.(pHE14), JM109(pHE16), JM109(pBD1), JM109(pHE18) and SURE.TM.(pML1) have been deposited Oct. 5, 1993 at BCCM at the Laboratorium voor Moleculaire Biologie (LMBP) (same address as LMG) and have the following accession numbers LMBP 2932, 2933, 2934, 2935 and LMBP 2936.

Growth of bacteria and phages. A. vinelandii was grown at 30.degree. C. with shaking in a nitrogen-free medium (9.8 mM K.sub.2 HPO.sub.4 /KH.sub.2 PO.sub.4, 0.8 mM MgSO.sub.4 7H.sub.2 O, 3.4 mM NaCl, 0.34 mM CaCl.sub.2, 8.7 .mu.M Na.sub.2 MoO.sub.4 2H.sub.2 O, 54 .mu.M FeSO.sub.4 7H.sub.2 O, 1% sorbitol, pH 7.4). E. coli was grown in LB-medium �Sambrook J, Fritsch, E. F. and Maniatis T., Molecular cloning, A laboratory manual, 2nd ed., Cold Spring Harbour Laboratory Press, New York, (1989)! with shaking at 37.degree. C. When the cells were to be used for growth of phages the LB-medium was supplemented with 2.5 mM CaCl.sub.2, 10 mM MgCl.sub.2, and 0.4% maltose. Phages were plated on strain Q359 on L-agar (LB-medium supplemented with 2% agar). Phage LB-medium supplemented with either 0.8% agar (titrations and gene library amplification) or 0.8% agarose (screening of gene library and preparation of phage lysates) was used for overlaying agar.

Standard recombinant DNA technology. Restriction endonuclease digestions, removal of cohesive DNA ends by using the 3' exonuclease activity of T4 DNA polymerase, ligations, agarose gel electrophoresis, and end-labelling with .sup.32 P were performed according to standard protocols �Sambrook J, Fritsch, E. F. and Maniatis T ., Molecular cloning, A laboratory manual, 2nd ed., Cold Spring Harbour Laboratory Press, New York, (1989)!. Transformations were performed as described by �Chung, C. T., Niemela S. L. and Miller R. H., Proc. Natl. Acad. Sci USA, 86, 2172-2175, (1989)!, and DNA sequencing was performed according to �Sanger F., Nicklen S., and Coulsom, A. R., Proc. Natl. Acad. Sci USA, 74, 4563 (1977)!.

Viscosimetric measurement. The alginate used in this experiment was obtained from Ascophyllum nodosum and had an intrinsic viscosity in 0.1 M NaCl of 17.6 dl/g at 25.degree. C. The viscosity was determined by an Ubbelhode viscosimeter.

NMR spectroscopy. The substrate used in these analyses was a low guluronic acid-containing alginate obtained from the brown algae Ascophyllum nodosum, and was prepared as described previously �Larsen, B., Proceedings of the Tenth International Seaweed Symposium, Ed: Levring, T.Gothenburg, p7-33, (1980)!. For the NMR analyses epimerase was obtained from IPTG-induced E. coli JM105 cells containing pHE5. 250 ml cell culture were harvested by centrifugation and resuspended in 20 ml of 10 mM Tris, 0.34 mM CaCl.sub.2, pH 7.0. After ultrasonication, the solution was centrifuged at 31.000.times.g for 1 hour. The supernatant was stored frozen at 70.degree. C. After thawing the supernatant was filtered through a membrane with pore size 0.22 .mu.m, and the enzyme was further purified on a Mono Q HR515 (Pharmacia) ion exchange column. The enzyme was eluted with a 0-1 M NaCl salt gradient (in the same buffer as the applied solution), and was collected in 2 ml at approximately 0.6 M NaCl. To each of two tubes was added 0.28 ml of this enzyme solution (0.9 mg/ml total protein), 1 ml alginate (7.5 mg/ml in H.sub.2 O), and 4.62 ml 2,3,6-trimethylpyridine buffer (see above). CaCl.sub.2 was then added to a total reaction volume of 6 ml such that one tube contained 0.85 .mu.M, and one contained 3.4 mM CaCl.sub.2. After incubation at 30.degree. C. for 20 hours Na.sub.2 EDTA (10 mM) was added to chelate the Ca.sup.2+ -ions, and the solutions were then dialyzed extensively against distilled water. The dialyzed alginate solutions were freeze-dried and then dissolved in D.sub.2 O. NMR spectroscopy of these solutions were finally performed according to �Grasdalen H., Larsen B., and Smidsr.o slashed.d O., Carbohydr. Res., 68, 23-31 (1979)! (Table 4). Further analysis was carried out in a similar fashion for DH5.alpha.(pHE8), JM109 (pHE16) and JM109(pBD9). The results in Table 4 conclusively demonstrate that the enzymatic activity is mannuronan C-5-epimerase activity. This activity is expressed from a number of the plasmids showing that an entire epimerase gene/protein is not required in order to maintain epimerase activity. The epimerase activity is Ca.sup.2+ dependent.

EXAMPLE 1

Purification of mannuronan C-5-epimerase (1), partial amino acid sequencing and synthesis of a mixed DNA probe. The enzyme was isolated from liquid cultures of A. vinelandii essentially as described in �Skj.ang.k-Br.ae butted.k, G. and Larsen, B. Carbohydrate Research, 103, (1982) 137-140!. The cells were removed by centrifugation and the enzyme was isolated by precipitation with 30% ammoniumsulphate and followed by centrifugation for 20 min. at 10000 rpm. The supernatant was then precipitated with 50% ammonium sulphate (final concentration), and the precipitate after centrifugation was dissolved in 0.05 M imidazole/HCl (pH 6.8) containing 0.34 mM CaCl.sub.2 and 0.5 mM dithiothreitol. This crude extract was then desalted on a prepacked column (PD-10) of Sephadex G-25 (Pharmacia) equilibrated with the same buffer. The extract was then applied on an alginate-Sepharose column. Proteins bound by non-specific interactions were eluted with 0.1 M NaCl. The epimerase was eluted as a sharp peak with 0.5 M NaCl. To make the enzyme pure enough for protein sequencing, it was dialyzed against TE-buffer overnight, freezedried, and further purified by SDS-PAGE gel electrophoresis (7.5% polyacrylamide in 25 mM Tris, 192 mM glycine, 0.1% sodium dodecyl sulphate, pH 8.3.) followed by electroblotting (in electrophoresis buffer without sodium dodecyl sulphate) onto a polyvinylidene difluoride membrane, poresize 0.45 .mu.m (Millipore). The membrane was stained with Coomassie brilliant blue and air dried, and the protein with Mw 122 kd was cut out for N-terminal sequencing on a model 477A protein sequencing apparatus from Applied Biosystems. A DNA oligonucleotide was synthesized on the basis of the amino acid sequence information, and this oligonucleotide was used as a probe for screening of the gene library after end-labelling with .sup.32 P by polynucleotide kinase.

EXAMPLE 2

Isolation of DNA from A. vinelandii and construction of a gene library. A. vinelandii cells were harvested and washed once in 0.9% Nacl. They were then lysed according to �Hansen, J. B. and Olsen, R. H., J. Bacteriol., 135, 227-238, (1978)!, and the lysate was extracted twice with phenol and twice with chloroform. Nucleic acids were precipitated with ethanol, and the DNA was collected on a glass rod and dissolved in TE-buffer (10 mM Tris, 1 mM Na.sub.2 EDTA, pH 7.9). Further purification was obtained by CsCl/ethidium bromide density gradient centrifugation. After removal of the ethidium bromide by isopropanol extraction, the DNA solution was dialyzed against TE buffer.

The DNA (molecular size greater than 60 kb) was subjected to partial Sau3AI digestion under conditions maximizing the generation of 15-20 kb fragments. After ethanol precipitation the DNA was dissolved in 40 .mu.l TE buffer to give a concentration of 0.5 .mu.g/.mu.l. The DNA was then dephosphorylated with calf intestine phosphatase, followed by inactivation of the enzyme by incubation at 75.degree. C. for 10 minutes in the presence of 10 nM nitrilotriacetic acid. The dephosphorylated DNA was precipitated with ethanol and dissolved in 40 .mu.l 0.1.times.TE buffer.

EMBL3 vector DNA was digested with BanHI+EcoRI, followed by an isopropanol precipitation step under conditions leaving the short BamHI/EcoRI oligonucleotides in solution �Frischauf A., Lehrach H., Poustka A. and Murray N., J. Mol. Biol., 170, 827-842, (1983)!. The Sau3AI-digested and dephosphorylated A. vinelandii DNA (1.75 .mu.g) was then ligated with the BamHI/EcoRI-digested vector DNA (4.75 .mu.g), using T4 DNA ligase in a total reaction volume of 20 .mu.l. After ligations over night at 10.degree. C., 10 .mu.l of the ligation mixture was subjected to in vitro packaging in a Promega Biotech packaging system. The in vitro constructed phage particles were titrated on the E. coli strain Q359, and the library was finally amplified on Q359 in one cycle by plating on solid medium. Screening of the library was performed according to standard protocols �Sambrook, J., Fritsch, E. F. and Maniatis, T., Molecular Cloning, Laboratory Manual, 2nd Ed., Cold Spring Harbour Laboratory Press, (1989)!, except that the highest stringency wash was 3.2 M tetramethylammonium chloride at 50.degree. C. A total of 1.4.times.10.sup.5 primary recombinant phages were constructed, a library complexity far above what is required for obtaining representativity of A. vinelandii genes.

EXAMPLE 3

Measurements of epimerase activity from mannuronan C-5-epimerase (1). (5-.sup.3 H) alginate was prepared as described in �Skj.ang.k-Br.ae butted.k, G. and Larsen, B., Carbohydrate Res., 103, 133-136, (1982)!. The (5-.sup.3 H)alginate was produced by growing Azotobacter vinelandii in a medium consisting of D-Glucose (20 g), K.sub.2 HPO.sub.4 (1 g), MgSO.sub.4.7 H.sub.2 O (200 mg), FeSO.sub.4. 7 H.sub.2 O (50 mg), NaMoO.sub.4. 2 H.sub.2 O (5 mg), NH.sub.4 OAc (2.3 g) and CaCl.sub.2. 2 H.sub.2 O (50 mg) diluted to one liter with water. The cells were grown at 30.degree. C. with vigorous shaking. After 30 hours, D-�5-.sup.3 H!glucose was added to a concentration of 0.6 mg/ml (Specific activity, 0.7 .mu.Ci/mg) and the cells were allowed to grow for another 72 hours. The culture was cooled in an ice-bath, and the cells were removed by centrifugation. The supernatant solution was dialysed against 0.05 M sodium EDTA (3.times.5 liters) for 24 hours followed by exhaustive dialysis against distilled water. The sodium alginate was then precipitated with ethanol in the presence of 0.2% of sodium chloride. The specific activity of the label was 29 000 dpm/mg alginate. The composition of this alginate was also analyzed by NMR spectroscopy, and it was found to contain 59% mannuronic acid. Phage lysates were prepared by plating 105 phages per plate. Two ml 2,3,6-trimethylpyridine buffer (50 mM, pH 6.9) were added to each plate, and the softagarose/buffer mixture was scraped off, vortexed and centrifuged at 10,000 rpm for 10 min. The supernatant was used for incubations with (5-.sup.3 H)alginate by mixing 0.25 ml (5-.sup.3 H)alginate (2.5 mg/ml), 6,3 .mu.l 0.1 M CaCl.sub.2, and 1.45 ml phage lysate. The mixture was incubated at 30.degree. C. overnight, and the alginate was precipitated by addition of 15 .mu.l 5 M NaCl and 2 ml ethanol. After incubation at -20.degree. C. for 30 min. the solution was centrifuged at 10.000 rpm for 30 min., and 1 ml of the supernatant was used for determination of released .sup.3 H �Skj.ang.k-Br.ae butted.k, G. and Larsen, B., Carbohydrate Res., 103, 133-136, (1982)!. in a liquid scintillation counter.

For measurements of epimerase activity as release of .sup.3 H in cells containing recombinant plasmids, the cell cultures were harvested by centrifugation and resuspended in 2,3,6 trimethylpyridine buffer. When IPTG (3 mM) was used for induction of the lac-promoter, the inducer was added to exponentially growing cells and incubations were continued for 3 hours. Cells were disrupted by ultra-sonication, and varying amounts of the lysates were incubated with shaking together with 100 .mu.l (5-.sup.3 H) alginate (2.5 mg/ml) and 400 .mu.l 2,3,6-trimethylpyridine buffer (total volume 0.6 ml) in the presence of 3.3 mM CaCl.sub.2. The quantities of enzyme-containing cell extracts used were adjusted such that the measurements were performed under conditions where the enzyme represented the limiting factor. After incubation at 30.degree. C. at the times indicated in each case, the mixtures were precipitated with ethanol under the conditions described above for phage lysates and 1.0 ml of the supernatant was used for scintillation counting. Controls (using the appropriate host with the pUC18 vector) gave low backgrounds and these numbers were subtracted in the values presented in Table 3.

EXAMPLE 4

Molecular cloning of a DNA fragment expressing a mannuronan C-5-epimerase activity in E. coli. The A. vinelandii gene library was constructed by cloning partially Sau3AI-digested A. vinelandii DNA into the bacteriophage .lambda. vector EMBL3. In order to identify the epimerase gene in this library, we constructed a DNA probe based on the assumption that the previously purified 122 kd protein represented the epimerase �Skj.ang.k-Br.ae butted.k, G. and Larsen, B., Carbohydr. Res., 103, 137-149 (1982)!. Initially we tried to use the corresponding protein solution for determination of the N-terminal amino acid sequence of the 122 kd protein, but the results showed that the preparation was not sufficiently pure for this purpose. We therefore purified the protein further by SDS-polyacrylamide gel-electrophoresis, followed by electroblotting onto a membrane. The band containing the 122 kd protein was cut out from this membrane and subjected to N-terminal amino acid sequence analysis. Based on parts of this sequence, we synthesized the mixed DNA probe shown in FIG. 1 (SEQ ID NO:6).

The DNA probe synthesized as in Example 1 was labelled with .sup.32 P and then used for screening of the A. vinelandii gene library. Clones which hybridized reproducibly against the labelled probe were identified at a frequency of approximately 10.sup.-3, and six such clones were selected for further studies. Phage lysates were prepared from each of the six clones, and each lysate was assayed for epimerase activity (Table 2). As can be seen, the lysates prepared from all six clones appeared to contain a weak enzyme activity that could represent the epimerase. This conclusion was further supported by the observation that control lysates prepared from randomly picked recombinant phages in the library, gave reproducibly lower activity, representing the background activity.

EXAMPLE 5

Subcloning of a DNA fragments encoding the epimerase. DNA from phage EP2 was partially digested with Sau3AI, and fragments ranging from 4 to 9 kb in size were subcloned in the BamHI site of plasmid pUC18. Cell extracts from DH5.alpha. transformants containing recombinant plasmids were assayed for epimerase activity, and the corresponding plasmids were also hybridized against the synthetic oligonucleotide used for screening of the gene library. The analysis of the cell extracts showed that one of them contained an enzymatic activity consistent with the assumption that a polypeptide having epimerase activity was expressed from the plasmid (pHE1) in this clone (see Table 3). We have also tried to centrifuge the extract at 30000 g for 3.5 hours, and found that the activity was not significantly reduced in the supernatant. Since we were unable to detect any significant activity in the culture medium, we conclude that the epimerase is localized intracellularly in E.coli. The insert in pHE1 also hybridized against the synthetic oligonucleotide used for screening, and pHE1 was therefore selected for further analysis.

EXAMPLE 6

Characterisation of the cloned DNA required for expression of the epimerase, and stability of the enzyme in vivo and in vitro. The insert in pHE1 is approximately 4 kb in size, and FIG. 2 shows the restriction map of this insert. Hybridization analysis of pHE1 with the original synthetic oligonucleotide showed that the sequence hybridizing to the oligonucleotide was localized downstream of the SphI site. The hybridizing sequence was further characterized by DNA sequencing, and this analysis showed that one of the potential reading frames of the sequence was in 100% agreement with the original N-terminal amino acid sequence of the 122 kd protein. Surprisingly, however, the orientation of the sequence was such that it would be transcribed out of the cloned fragment (see FIG. 2). This result thus indicated that the observed epimerase activity was not correlated with the sequence encoding the 122 kd protein, a conclusion that was further confirmed by the observation that the terminal 0.5 kb SphI fragment could be deleted (generating plasmid pHE7) without loss of the epimerase activity from the corresponding cell extract. In addition to the SphI deletion, we deleted (from pHE7) the 0.7 kb KpnI fragment at the opposite terminus of the insert, generating plasmid pHE5. As shown in Table 3, pHE5 (in DH5.alpha.) expressed the epimerase at a level approximately 27 times higher than the level of expression from pHE1.

During the expression studies described above we found that the measurements were quantitatively difficult to reproduce unless the time of harvesting the cells were kept as constant as possible. We have analyzed this problem more carefully by measuring the enzyme activity at different stages of growth of the E. coli cells. The results of such an analysis are shown in FIG. 3, and indicate that the enzymatic activities in the cell extracts are drastically reduced shortly after the cells have entered the stationary phase. To obtain optimal enzyme yields it is therefore important to harvest the cells at the end of the exponential phase or at the beginning of the stationary phase. The reason for the reduction of epimerase activity might potentially be due to proteolysis of the epimerase in stationary phase cells. To study the stability of the enzyme in vitro we have also analyzed the kinetics of .sup.3 H release in the DH5.alpha.(pHE5) extracts. As can be seen from FIG. 4, the enzyme activity is linear over at least 30 hours, demonstrating that the enzyme is very stable in vitro. A critical parameter for obtaining reproducible results is thus the time of harvesting of the cells.

EXAMPLE 7

Stimulation of the epimerase activity by induction of the lac promoter. The results described above showed that the levels of expression of the epimerase from pHE5 was significantly higher than in pHE1. The reasons for this could potentially be that the lac promoter was important for the expression, and we have therefore analysed this problem more closely. The analyses were performed in the E. coli strain JM105, a strain which expresses high levels of lac repressor, thus allowing a more repressed state of the promoter under uninduced conditions. When cell extracts prepared from uninduced and induced (with IPTG) cells of JM105(pHE1), a significant stimulation of enzyme activity was observed in the induced cells (Table 3). A similar experiment using JM105(pHE5) showed even greater stimulation of the expression of the epimerase upon addition of IPTG in this case. These experiments thus showed that the lac promoter probably is a key element, although not necessarily the only element, involved in the expression of the epimerase from pHE5. The experiments in addition showed that the direction of transcription is from the KpnI towards the SphI site in the insert. The epimerase gene is therefore transcribed in the same direction as the gene encoding the 122 kd protein, whose N-terminal amino acid sequence was used for the isolation of the cloned DNA.

Preliminary experiments on deleting more DNA from the SphI side of the insert indicated that very little could be deleted without loss of the epimerase activity. At the KpnI side, on the other hand, we found that significant deletions were tolerated. Table 3 shows the results of analysis of expression of the epimerase from a plasmid (pHE8) constructed by deleting the 0.8 kb KpnI/SacII fragment from pHE5. As can be seen, this deletion resulted in a very strong stimulation of the epimerase activity both in uninduced and induced cells. The expression from pHE8 is presumably based on initiation of translation from the Shine-Dalgarno sequence in the vector (localized between the lac promoter and the polylinker). Similarly, high levels of expression were obtained from pHE22 also due to the coding sequences being in frame with the Shine-Dalgano sequence. So far we have not obtained expression of the epimerase in constructs where deletions have extended beyond the SacII site.

EXAMPLE 8

Use of a different promotor than the lac promotor. The insert in pHE5 (EcoRI-HindIII) was sub-cloned into plasmid pT7-3 (a derivative of pT7-1 described by �Tabor, S., and C. C. Richardson (1985). Proc. Natl. Acad. Sci. 82, 1074-1078!), and the new plasmid was designated pLB1. The insert in pLB1 is localized downstream of the .phi.10 promoter in the vector. This promoter is only recognized by the bacteriophage T7 RNA polymerase, and expression of genes downstream of this promoter thereby becomes dependent on expression of this polymerase activity in the cells. The 442 bp (see FIG. 2) SacI- SpoI fragment was finally deleted from the insert in pLB1, generating plasmid pLB2. pLB2 was transformed into E. coli K38 (pGP1-2). Plasmid pGP1-2 encodes the gene for T7 RNA polymerase, and the expression of the gene is controlled by a temperature inducible repressor. K38(pLB1, pGP1-2) was grown in exponential phase at 30.degree. C. for 41/2 hours. One of two parallel cell cultures was then transferred to 42.degree. C. for 30 minutes to induce the T7-polymerase. The other parallel cell culture was grown at 30.degree. C. for 5 hours. The epimerase activities in the cells were measured as described in example 3, and the results of the measurements are shown in Table 3.

EXAMPLE 9

Cloning of mannuronan C-5-epimerase (2). Plasmid pHE12 was constructed by inserting a 6.2 kb XhoI fragment from the recombinant bacteriophage lambda derivative EP2 into pUC128. As can be seen from FIG. 2 the insert in pHE12 is partly overlapping with the insert in pHE1. Analysis of extracts prepared from cells containing pHE12 (as described for pHE1), showed that they expressed mannuronan C-5-epimerase activity (Table 3). Further analysis showed that the 2.5 kb SpoI-XhoI fragment could be deleted from the insert in pHE12 without affecting the expression of mannuronan C-5-epimerase. Further plasmids were constructed (see FIG. 2) and the activity analysed (see Table 3). This demonstrated that both the genes and gene fragments were able to express epimerase activity. The nucleotide sequences of the inserts were determined by the method of Sanger �Sanger, F., S. Nicklen, and A. R. Coulson. 1977. Proc. Natl. Acad. Sci. 74, 5436!. The nucleotide sequences are shown in FIG. 6.

EXAMPLE 10

Sequence Comparison. Five genes have been identified as shown in FIG. 2. The insert containing E5 is located about 5-10 kilobases away from the other genes. SEQ ID NO:1 shows the nucleotide sequence for the complete genes of E4, E1, E2 and a large portion of E3. SEQ ID NO:2-SEQ. ID NO:5 sequentially show the amino acid sequences for E4, E1, E2 and a large portion of E3. Detailed analysis of the nucleotide and amino acid sequences revealed highly homologous regions within each gene and between the various genes. FIG. 5 characterises each of the genes by reference to the homologous blocks. Each of the genes has at least one A-element and at least one R-element.

E1, E2 and E4 all end with a reasonably homologous sequence termed the S-element (not shown in FIG. 5). The last 14 amino acids of the S-element of E1 and E2 are identical with one exception. FIGS. 6-9 show detailed analysis of the A- and R-elements within each gene by reference to the consensus sequence (con) (SEQ ID NO:7-SEQ ID NO:10). Each A-element is approximately 1,150 base pairs long and each R-element is approximately 450 base pairs long. Short oligonucleotides are present in E1, E2 and E3 between the second and third R-elements. Gaps have been introduced where necessary to maximise alignment (see in particular the third R-element of E2).

Hybridization with a probe made from the first part of the A-element to a Southern blot of A. vinelandii digested with restriction endonuclease Bg1II gave 5 distinct bands. One of these bands contained two A blocks, and another of these bands contained two different fragments with the same size. The number of bands were the same when other strains (ATCC 478, ATCC 12837 and ATCC 12518) of the same species were used. This implies that the bacterium contains at least 5 copies of the A-element, and that this is common to several independently isolated strains of A. vinelandii.

The first part of each R-element contains six perfect and imperfect repeats of a nonapeptide with the consensus sequence LXGGAGXDX (SEQ ID NO:11), except for the third R-element of E2 which lacks two of these repeats. FIG. 11 shows the complete nucleotide (SEQ ID NO:1) and corresponding amino acid sequence of E2. The nonapeptides have been marked with double lines for a good match with the consensus sequence and single lines for less good matches. This nonapeptide motif is characteristic of the haemolysin family of secreted proteins (Suh, Y. and Benedik, M. J., J. Bacteriol 174, (1992) 2361-2366). These proteins are all calcium dependent and are secreted by a pathway which does not involve cleavage of an N terminal signal peptide. For haemolysin secreted from E. coli it has been proposed that the nonamers are responsible for the binding of calcium ions (Ludwig, A. et al, Mol. Gen. Genet. 214, (1988) 553-561, Boehm, D. F. et al, Infect. Immun. 58 (1990) 1959-1964). It appears that the R-elements are involved in calcium ion binding, calcium being necessary for both enzyme activity and gel formation.

EXAMPLE 11

Making an altered epimerase.

As can be seen from Table 3, various elements may be deleted from the gene, while maintaining the expression of a protein having epimerase activity. Clearly, the latter portion of E1 having the sequence ARS has epimerase activity (see plasmid pHE8), although deletions in A2 of E1 are not tolerated (see Example 7). Additionally, E2 having the sequence ARRRRS (SEQ ID NO:12) also demonstrates epimerase activity. Additionally, fragments of E3 lacking a carboxy terminal and having the sequences ARRR (SEQ ID NO:15) and ARRRARR (SEQ ID NO:13) have expressed epimerase activity. (See plasmids pH18 and pBD6). Accordingly, it appears that an S-element is not essential for epimerase activity, although the presence of this element may affect activity. We therefore postulated that an epimerase may need at least one A-element and at least one R-element, and that it should be possible to make altered epimerases by combining these elements in different ways. To show this, we constructed a plasmid encoding an epimerase with the sequence RARS (SEQ ID NO:14):

The insert in pHE1 (EcoRI-HindIII) was subcloned into plasmid pTrc99A (Pharmacia), generating plasmid pHE21. This plasmid contains a trc-promoter in front of the epimerase I gene, a strong transcription termination signal downstream of the gene, and the lacI.sup.q -gene allowing induction with IPTG. pHE21 was digested with KpnI and SpoI, made blunt-ended with S1 nuclase and religated. The resulting plasmid, pHE22, expresses a protein having the carboxy terminal of epimerase 1, RARS. The epimerase activity was measured as in Example 3, see Table 3.

Given that epimerase activity is expressed from a number of the constructs, it seems likely that a number of synthetic enzymes may be produced having epimerase activity including differing numbers of A, R and S blocks. The presence of activity in pHE22 implies that it is not essential to have an amino-terminal A block, and so block order may also be altered.

EXAMPLE 12

The .sup.1 H-NMR spectra of alginate epimerased by extracts from plasmids pHE8 and pBD9 show that the proteins encoded by these plasmids have different enzyme activity, pHE8 producing epimerase with single G activity while pBD9 producing epimerase with G block activity. pHE8 encodes the carboxy terminal ARS of E1 whereas pBD9 encodes ARRRRS (SEQ ID NO:12) of E2. The naturally encoded epimerases may therefore have differing activity particularly in the distribution patterns of Gs. The different activity of the various epimerases encoded within the 5 genes could be used to create alignates having a desired structure, by selectively expressing a desired gene or genes. Alternatively, it may be possible to construct synthetic enzymes varying the A, R and S block content of each epimerase to provide enzymes having altered activity providing a further level of control in the production of desired alginates.

TABLE 1______________________________________Bacterial strains, phages, and plasmidsStrain/phage/plasmid Remarks References______________________________________BacterialstrainsA. vinelandii Strain E Larsen and Haug (1971)E. coliQ359 supE hsdR .phi.80.sup..left brkt-top. P2 Karn et al. (1980)DH5.alpha. supE44.DELTA.lacU169 (.phi.80 lacZ.DELTA.M15) Bethesda Research hsdR17 recA1 endA1 gyrA96 Laboratories (1986) thi-1 relA1JM105 supE endA sbcB15 hsdR4 rpsL Yanisch-Perron thi.DELTA.(lac--proAB) et al (1985)JM109 recA1 supE44 endA1 hsdR17 Yanisch-Perron gyrA96 relA1 thi.DELTA.(lac--proAB) et al (1985) F�traD36 proAB .sup.+ lacI.sup.q lacZ.DELTA.M15!SURE .TM. e14.sup.- (mcrA), Greener (1990) .DELTA.(mcrCB--hsdSMR--mrr) 171, endA1, supE44, thi-1, gyrA96, relA1, lac, recB, recJ, sbcC, umuC:Tn5(kan.sup..left brkt-top.), uvrC, �F' proAB, lacI.sup.q Z.DELTA.M15, Tn10, (tet.sup..left brkt-top.)!.PhagesEMBL3 Bacteriophage .lambda. vector used for Frischauf et al. construction of A. vinelandii (1983) gene libraryEPx Randomly picked phage from See examples A. vinelandii gene libraryEP2, -3, Phages isolated from A. See examples-6, -7, vinelandii gene library-8 and -9 Identified by hybridization and expresses mannuronan C-5-epimerasePlasmidspUC18 Ampicillin resistance, ColE1 Norrander et al. repliconpUC128 Ampicillin resistance, ColE1 Keen et al. (1988) repliconpTrc99A Ampicillin resistance, ColE1 Pharmacia repliconpT7-3 Ampicillin resistance, ColE1 Tabor & Richardson replicon (1985)pGP1-2 Kanamycin resistance, P15A Tabor & Richardson replicon (1985)pBluescript Ampicillin resistance, ColE1II SK(+) repliconpHE1 Derivative of pUC18 where a See examples 4 kb Sau3A1 DNA fragment from phage EP2 was subcloned into the BamH1 polylinkerpHE7 Derivative of pHE1 where a 0.5 kb See examples Sph1 DNA fragment was deletedpHE5 Derivative of pHE7 where a 0.7 kb See examples Kpn1 DNA fragment was deletedpHE8 Derivative of pHE5 where a 0.8 kb See examples Kpn1/SacII DNA fragment was deleted. Cohesive ends were removed prior to ligation by using the 3' exonuclease activity of T4 DNA polymerasepLB1 Derivative of pT7-3, where the See examples 2,7 kb insert from pHE5 was cloned into the EcoRI/HindIII polylinkerpLB2 Derivative of pLB1, where a 0.4 kb See examples SacI/SpoI fragment was deletedpHE12 Derivative of pUC128, where a See examples 6.2 kb XhoI fragment from phage EP2 was subcloned into the XhoI polylinkerpBD1 Derivative of pHE12, where a See examples 2.0 kb SpoI/NsiI fragment was deletedpHE21 Derivative of pTrc99A, where the See examples 4.0 kb insert of pHE1 was cloned into the EcoRI/HindIII polylinkerpHE22 Derivative of pHE21, where a See examples 1.2 kb KpnI/SpoI fragment was deletedpHE2 Derivative of pUC18 where a See examples 6.0 kb SphI DNA fragment from phage EP6 was cloned into the SphI site in the polylinker of the vectorpHE16 Derivative of pHE2 where a See examples 1.5 kb EcoRI--SmaI DNA fragment was deletedpBD9 Derivative of pBD1 where a See examples 0.4 kb XhoI--FspI DNA fragment was deletedpBD6 Derivative of pHE12 where a See examples 3.4 kb XhoI--EspI DNA fragment was deletedpHE18 Derivative of pUC128 where an See examples 5.1 kb NotI--PvuII DNA fragment from EP6 was cloned into the NotI--EcoRV sites in the polylinker of the vectorpHE14 Derivative of pUC128 where a See examples 3.0 kb BglII DNA fragment from EP6 was cloned into the BamHI site of the polylinkerpML1 Derivative of pBluescript II See examples SK(+) where a 4.3 kb KpnI--SacII DNA fragment was cloned into the corresponding sites in the polylinker______________________________________ References to Table 1 Larsen, B., and A. Haug. 1971. Biosynthesis of alginate. Carbohydr. Res. 17:287-296. Karn, J., S. Brenner., L. Barnett, and G. Cesareni. 1980. Proc. Natl. Acad. Sci. U.S.A. 77:5172. Bethesda Research Laboratories. 1986. Bethesda Res. Lab. Focus 8(2):9. Yanisch-Perron, C., J. Vieira, and J. Messing. 1985. Gene 33:103-119. Frischauf, A., H. Lehrach, A. Poustka, and N. Murray. 1983. J. Mol. Biol. 170:827-842. Norrander, J., T. Kempe, J. Messing. 1983. Gene 26:101-106. Keen N. T, S. Tamaki, D. Kobayashi and D. Trollinger (1988). Gene 79:191-197. Tabor S. and C. C. Richardson (1985). Proc Natl Acad Sci USA 82:10741078. Greener, A. (1990) Strategies 3:5-6. TABLE 2______________________________________Putative mannuronan C-5-epimerase activityin recombinant phage lysates.Recombinant phage .sup.3 H release (dpm)______________________________________EPx 39EP2 91EP3 107EP6 74EP7 75EP8 245EP9 75______________________________________ EPx originated from the A. vinelandii gene library as a randomly picked plaque, while the other six phages were selected on the basis of the hybridization between their DNA and the labelled oligonucleotide used for screening of the library. TABLE 3______________________________________Activity of the mannuronan C-5-epimeraseexpressed from the plasmids. Released .sup.3 H/0D.sub.600 unit cell cultureEnzyme Strain No IPTG IPTG______________________________________Epimerase 1.sup.1 JM109 (pHE16) 10000 11000Epimerase 1.sup.2 DH5.alpha. (pHE1) 273 ndEpimerase 1.sup.3 DH5.alpha. (pHE5) 9700 ndEpimerase 1.sup.4 JM105 (pHE1) 637 2800Epimerase 1.sup.5 JM105 (pHE5) 4800 28500Epimerase 1.sup.6 JM105 (pHE8) 58900 181000Epimerase 1.sup.7 JM109 (pHE21) 93 1283Epimerase 1.sup.9 JM109 (pHE22) 5383 34611Epimerase 1.sup.9 K38 (pGP1-2, pLB2)* 2150 8333Epimerase 2.sup.1 JM109 (pHE12) nd 140Epimerase 2.sup.2 JM109 (pBD9) nd 6700Epimerase 3.sup.1 JM109 (pHE18) 551 2270Epimerase 3.sup.2 JM109 (pBD6) nd 530Epimerase 4 DH5.alpha. (pHE14) nd 3500______________________________________ The extracts were incubated with the alginate for 16 hours, and the numbers are given in dpm. nd = not determined. *Thge culture was not induced by IPTG, but by raising the temperature fro 30.degree. C. to 42.degree. C. TABLE 4______________________________________NMR analysis of the reaction product after incubation of therecombinant epimerase with a low guluronic acid containing substrate CaCl.sub.2 Frequences of M and G residuesStrain (mM) F.sub.M F.sub.G F.sub.MM F.sub.MG F.sub.GG F.sub.GG /F.sub.G______________________________________JM105(pHE5) 0.85 0.80 0.20 0.66 0.14 0.06 0.3JM105(pHE5) 3.4 0.71 0.29 0.45 0.26 0.03 0.10DH5.alpha.(pHE8) 3.4 0.72 0.28 0.52 0.20 0.08 0.29JM109(pHE16)* 3.4 0.82 0.18 0.79 0.03 0.15 0.83JM109(pBD9) 2.1 0.60 0.40 0.54 0.06 0.34 0.85______________________________________ *This spectrum was obtained at a 400 MHz instrument to be able to get relatively correct figures in spite of the lower conversion. __________________________________________________________________________# SEQUENCE LISTING- (1) GENERAL INFORMATION:- (iii) NUMBER OF SEQUENCES: 52- (2) INFORMATION FOR SEQ ID NO:1:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 12588 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (vi) ORIGINAL SOURCE:#vinelandiiA) ORGANISM: Azotobacter (B) STRAIN: E- (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 290..1951- (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 2227..6438- (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 6702..9695- (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 9973..12588- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:- GATCCGGCCG TCTGAGACGG CGCCTCCGGC CGTCGGCGAG TGCGCCGTTC GC - #CGACGGCC 60- GGGCGAACGG ATGAGGACTG CTCCACTCTC ACCCAGATAA GCGCGTGGGC CG - #TTTCATCC 120- GAGCGCCTTT CCGGGCCGCT TCGAAAGACC GCCACGAGGC ACTCTGTGCA AG - #GGCCAGGC 180- AGTCGCGTTG CAACCGGAGA CGGGACCGGC CCGTTCGGGC GTCGTCTCTT CC - #CGCTCCAC 240- TTTTTCCAGG CAGCTTCGGC TGCTCCACTC GGAACCGGGA AGCGGAGAT ATG - # GAT 295#Met Asp# 1- TAC AAC GTC AAG GAT TTC GGT GCA TTG GGC GA - #C GGC GTC AGC GAC GAC 343Tyr Asn Val Lys Asp Phe Gly Ala Leu Gly As - #p Gly Val Ser Asp Asp# 15- CGG GCC TCC ATC CAG GCG GCG ATC GAT GCC GC - #C TAC GCC GCC GGT GGC 391Arg Ala Ser Ile Gln Ala Ala Ile Asp Ala Al - #a Tyr Ala Ala Gly Gly# 30- GGT ACC GTC TAC CTG CCG GCC GGC GAG TAC CG - #G GTC AGC GCC GCC GGG 439Gly Thr Val Tyr Leu Pro Ala Gly Glu Tyr Ar - #g Val Ser Ala Ala Gly# 50- GAG CCG GGC GAC GGC TGC CTG ATG CTC AAG GA - #C GGC GTC TAC CTG GCC 487Glu Pro Gly Asp Gly Cys Leu Met Leu Lys As - #p Gly Val Tyr Leu Ala# 65- GGT GCC GGC ATG GGC GAG ACG GTG ATC AAG CT - #G ATC GAC GGC TCC GAC 535Gly Ala Gly Met Gly Glu Thr Val Ile Lys Le - #u Ile Asp Gly Ser Asp# 80- CAG AAG ATC ACC GGC ATG GTC CGC TCG GCC TA - #C GGC GAG GAA ACC AGC 583Gln Lys Ile Thr Gly Met Val Arg Ser Ala Ty - #r Gly Glu Glu Thr Ser# 95- AAC TTC GGC ATG CGC GAC CTG ACC CTC GAC GG - #C AAC CGC GAC AAC ACC 631Asn Phe Gly Met Arg Asp Leu Thr Leu Asp Gl - #y Asn Arg Asp Asn Thr# 110- AGC GGC AAG GTC GAC GGC TGG TTC AAC GGC TA - #T ATC CCC GGC GGG GAC 679Ser Gly Lys Val Asp Gly Trp Phe Asn Gly Ty - #r Ile Pro Gly Gly Asp115 1 - #20 1 - #25 1 -#30- GGC GCC GAC CGC GAC GTG ACC ATC GAG CGG GT - #G GAG GTC CGC GAG ATG 727Gly Ala Asp Arg Asp Val Thr Ile Glu Arg Va - #l Glu Val Arg Glu Met# 145- TCC GGC TAC GGC TTC GAC CCC CAC GAG CAG AC - #C ATC AAC CTG ACG ATC 775Ser Gly Tyr Gly Phe Asp Pro His Glu Gln Th - #r Ile Asn Leu Thr Ile# 160- CGC GAC AGC GTG GCC CAC GAC AAC GGC CTC GA - #C GGC TTC GTC GCC GAC 823Arg Asp Ser Val Ala His Asp Asn Gly Leu As - #p Gly Phe Val Ala Asp# 175- TAC CTG GTC GAC AGC GTG TTC GAG AAC AAC GT - #C GCC TAC GCC AAC GAC 871Tyr Leu Val Asp Ser Val Phe Glu Asn Asn Va - #l Ala Tyr Ala Asn Asp# 190- CGC CAC GGC TTC AAC GTG GTC ACC AGC ACC CA - #C GAT TTC GTC ATG ACC 919Arg His Gly Phe Asn Val Val Thr Ser Thr Hi - #s Asp Phe Val Met Thr195 2 - #00 2 - #05 2 -#10- AAC AAC GTC GCC TAC GGC AAC GGC AGC AGC GG - #C CTG GTG GTG CAG CGG 967Asn Asn Val Ala Tyr Gly Asn Gly Ser Ser Gl - #y Leu Val Val Gln Arg# 225- GGT CTG GAG GAC CTC GCG CTG CCC AGC AAC AT - #C CTG ATC GAC GGC GGC1015Gly Leu Glu Asp Leu Ala Leu Pro Ser Asn Il - #e Leu Ile Asp Gly Gly# 240- GCC TAC TAC GAC AAC GCC CGC GAA GGC GTG CT - #G CTC AAG ATG ACC AGC1063Ala Tyr Tyr Asp Asn Ala Arg Glu Gly Val Le - #u Leu Lys Met Thr Ser# 255- GAC ATC ACC CTG CAG AAC GCC GAT ATC CAC GG - #C AAC GGC TCC TCC GGG1111Asp Ile Thr Leu Gln Asn Ala Asp Ile His Gl - #y Asn Gly Ser Ser Gly# 270- GTG CGC GTC TAC GGC GCC CAG GAC GTG CAG AT - #C CTC GAT AAC CAG ATC1159Val Arg Val Tyr Gly Ala Gln Asp Val Gln Il - #e Leu Asp Asn Gln Ile275 2 - #80 2 - #85 2 -#90- CAC GAC AAC GCG CAG GCG GCC GCC GTG CCC GA - #G GTC CTG CTG CAG TCC1207His Asp Asn Ala Gln Ala Ala Ala Val Pro Gl - #u Val Leu Leu Gln Ser# 305- TTC GAC GAT ACC GCC GGG GCG TCC GGC ACC TA - #C TAC ACG ACC CTG AAC1255Phe Asp Asp Thr Ala Gly Ala Ser Gly Thr Ty - #r Tyr Thr Thr Leu Asn# 320- ACC CGG ATC GAG GGC AAC ACC ATC AGC GGC TC - #G GCC AAC TCC ACC TAC1303Thr Arg Ile Glu Gly Asn Thr Ile Ser Gly Se - #r Ala Asn Ser Thr Tyr# 335- GGC ATC CAG GAG CGC AAC GAC GGC ACC GAC TA - #C AGC AGC CTG ATC GAC1351Gly Ile Gln Glu Arg Asn Asp Gly Thr Asp Ty - #r Ser Ser Leu Ile Asp# 350- AAC GAC ATC GCC GGG GTG CAA CAG CCC ATC CA - #A CTG TAC GGA CCT CAC1399Asn Asp Ile Ala Gly Val Gln Gln Pro Ile Gl - #n Leu Tyr Gly Pro His355 3 - #60 3 - #65 3 -#70- TCG ACG GTA TCC GGC GAA CCC GGC GCG ACA CC - #G CAA CAG CCG TCC ACG1447Ser Thr Val Ser Gly Glu Pro Gly Ala Thr Pr - #o Gln Gln Pro Ser Thr# 385- GGA AGC GAC GGC GAG CCA CTG GTC GGC GGC GA - #C ACG GAC GAC CAG CTC1495Gly Ser Asp Gly Glu Pro Leu Val Gly Gly As - #p Thr Asp Asp Gln Leu# 400- CAG GGC GGC TCC GGC GCC GAT CGC CTG GAC GG - #C GGG GCC GGC GAC GAC1543Gln Gly Gly Ser Gly Ala Asp Arg Leu Asp Gl - #y Gly Ala Gly Asp Asp# 415- ATC CTC GAC GGC GGC GCC GGG CGC GAC CGG CT - #G AGC GGC GGC GCG GGC1591Ile Leu Asp Gly Gly Ala Gly Arg Asp Arg Le - #u Ser Gly Gly Ala Gly# 430- GCC GAC ACC TTC GTG TTC TCC GCC CGC GAG GA - #C AGC TAC CGT ACC GAC1639Ala Asp Thr Phe Val Phe Ser Ala Arg Glu As - #p Ser Tyr Arg Thr Asp435 4 - #40 4 - #45 4 -#50- ACG GCG GTG TTC AAC GAC CTG ATC CTC GAC TT - #C GAG GCC AGC GAG GAT1687Thr Ala Val Phe Asn Asp Leu Ile Leu Asp Ph - #e Glu Ala Ser Glu Asp# 465- CGC ATC GAC CTG TCC GCG CTG GGC TTT TCC GG - #C CTG GGC GAC GGC TAT1735Arg Ile Asp Leu Ser Ala Leu Gly Phe Ser Gl - #y Leu Gly Asp Gly Tyr# 480- GGC GGC ACC CTG CTC CTG AAG ACC AAC GCC GA - #G GGC ACG CGC ACC TAC1783Gly Gly Thr Leu Leu Leu Lys Thr Asn Ala Gl - #u Gly Thr Arg Thr Tyr# 495- CTG AAA AGC TTC GAG GCG GAT GCC GAG GGA CG - #G CGC TTC GAG GTC GCC1831Leu Lys Ser Phe Glu Ala Asp Ala Glu Gly Ar - #g Arg Phe Glu Val Ala# 510- CTG GAC GGC GAC CAC ACG GGC GAT CTT TCC GC - #C GCC AAT GTG GTC TTC1879Leu Asp Gly Asp His Thr Gly Asp Leu Ser Al - #a Ala Asn Val Val Phe515 5 - #20 5 - #25 5 -#30- GCC GCG ACC GGG ACG ACC ACC GAA CTC GAA GT - #G CTC GGC GAC AGC GGC1927Ala Ala Thr Gly Thr Thr Thr Glu Leu Glu Va - #l Leu Gly Asp Ser Gly# 545- ACG CAG GCC GGG GCG ATC GTC TAG CGCGTCCCGC TC - #CGACACAT AGCCGGTCGT1981Thr Gln Ala Gly Ala Ile Val * 550- CGGCAAGGCG GCCGGCCGCC GGCTGCCCCG AAGTTTCCAA TCTAATCTCA CC - #TACAGACA2041- GGCGCGTTCC GGTGCGCCCG AGCGCCGCCC CCGGGAACGA CCGGCAGGGC GT - #GTTTGTGC2101- GCAAGGTGCA GGCGGTCGCG CTCGAAGCCA GAGGCAGGGA AAACCTTTTC CG - #GCAGTCGT2161- CTCTTCCTTC TCCACTTCCC AGGCAGCCCT GGGCCGAGCA ACACGACGGG AT - #TAGGAAGC2221#GCA CTG GGC GAT GGC 2268 GAT TTC GGA#Phe Gly Ala Leu Gly Asp Glys Asp# 10- GTC AGC GAC GAC ACG GCG GCC ATC CAG GCG GC - #G ATC GAC GCC GCC CAC2316Val Ser Asp Asp Thr Ala Ala Ile Gln Ala Al - #a Ile Asp Ala Ala His# 30- GCG GCG GGC GGC GGC ACC GTC TAC CTG CCG GC - #C GGC GAA TAT CGG GTC2364Ala Ala Gly Gly Gly Thr Val Tyr Leu Pro Al - #a Gly Glu Tyr Arg Val# 45- AGC GGC GGC GAG GAG CCT TCC GAT GGT TGT CT - #G ACC ATC AAG AGC AAC2412Ser Gly Gly Glu Glu Pro Ser Asp Gly Cys Le - #u Thr Ile Lys Ser Asn# 60- GTC CAT ATC GTC GGC GCC GGG ATG GGC GAG AC - #G GTG ATC AAG ATG GTC2460Val His Ile Val Gly Ala Gly Met Gly Glu Th - #r Val Ile Lys Met Val# 75- GAC GGC TGG ACG CAG AAC GTC ACC GGC ATG GT - #G CGC TCG GCC TAC GGC2508Asp Gly Trp Thr Gln Asn Val Thr Gly Met Va - #l Arg Ser Ala Tyr Gly# 90- GAG GAA ACC AGC AAC TTC GGC ATG AGC GAC CT - #G ACC CTC GAC GGC AAC2556Glu Glu Thr Ser Asn Phe Gly Met Ser Asp Le - #u Thr Leu Asp Gly Asn#110- CGC GAC AAC CTG TCC GCC AAG GTC GAC GGC TG - #G TTC AAC GGC TAC ATC2604Arg Asp Asn Leu Ser Ala Lys Val Asp Gly Tr - #p Phe Asn Gly Tyr Ile# 125- CCC GGC CAG GAC GGC GCC GAT CGC GAC GTG AC - #C CTG GAG CGG GTG GAA2652Pro Gly Gln Asp Gly Ala Asp Arg Asp Val Th - #r Leu Glu Arg Val Glu# 140- ATC CGC GAG ATG TCC GGC TAC GGT TTC GAC CC - #C CAC GAG CAG ACC ATC2700Ile Arg Glu Met Ser Gly Tyr Gly Phe Asp Pr - #o His Glu Gln Thr Ile# 155- AAC CTG ACG ATC CGC GAC AGC GTG GCC CAC GA - #C AAC AGC CTC GAC GGC2748Asn Leu Thr Ile Arg Asp Ser Val Ala His As - #p Asn Ser Leu Asp Gly# 170- TTC GTC GCC GAC TAC CAG GTC GGC GGG GTG TT - #C GAG AAC AAC GTC TCG2796Phe Val Ala Asp Tyr Gln Val Gly Gly Val Ph - #e Glu Asn Asn Val Ser175 1 - #80 1 - #85 1 -#90- TAC AAC AAC GAC CGC CAC GGC TTC AAC ATC GT - #C ACC AGC ACC AAC GAC2844Tyr Asn Asn Asp Arg His Gly Phe Asn Ile Va - #l Thr Ser Thr Asn Asp# 205- TTC GTC CTG AGC AAC AAC GTC GCC TAC GGC AA - #C GGC GGC GCC GGC CTG2892Phe Val Leu Ser Asn Asn Val Ala Tyr Gly As - #n Gly Gly Ala Gly Leu# 220- GTG GTG CAG CGC GGC TCG TAC GAC CTG CCC CA - #T CCC TAC GAC ATC CTG2940Val Val Gln Arg Gly Ser Tyr Asp Leu Pro Hi - #s Pro Tyr Asp Ile Leu# 235- ATC GAC GGC GGC GCC TAC TAC GAC AAC GCC TT - #G GAA GGC GTG CAG CTC2988Ile Asp Gly Gly Ala Tyr Tyr Asp Asn Ala Le - #u Glu Gly Val Gln Leu# 250- AAG ATG GCC CAC GAC GTC ACC CTG CAG AAC GC - #C GAG ATC TAC GGC AAC3036Lys Met Ala His Asp Val Thr Leu Gln Asn Al - #a Glu Ile Tyr Gly Asn255 2 - #60 2 - #65 2 -#70- GGC CTG TAC GGG GTG CGC GTC TAC GGC GCC CA - #G GAC GTG CAG ATC CTC3084Gly Leu Tyr Gly Val Arg Val Tyr Gly Ala Gl - #n Asp Val Gln Ile Leu# 285- GAC AAC CAG ATC CAC GAC AAT TCG CAG AAC GG - #C GCC TAT GCC GAA GTC3132Asp Asn Gln Ile His Asp Asn Ser Gln Asn Gl - #y Ala Tyr Ala Glu Val# 300- CTG CTG CAG TCC TAC GAC GAC ACC GCC GGG GT - #G TCC GGC AAC TTT TAC3180Leu Leu Gln Ser Tyr Asp Asp Thr Ala Gly Va - #l Ser Gly Asn Phe Tyr# 315- GTC ACC ACC GGC ACC TGG CTC GAA GGC AAC GT - #C ATC AGC GGC TCG GCC3228Val Thr Thr Gly Thr Trp Leu Glu Gly Asn Va - #l Ile Ser Gly Ser Ala# 330- AAT TCC ACC TAC GGC ATC CAG GAG CGC GCC GA - #C GGC ACC GAC TAC AGC3276Asn Ser Thr Tyr Gly Ile Gln Glu Arg Ala As - #p Gly Thr Asp Tyr Ser335 3 - #40 3 - #45 3 -#50- AGC CTC TAC GCC AAC AGC ATC GAC GGT GTG CA - #G ACC GGG GCG GTA CGG3324Ser Leu Tyr Ala Asn Ser Ile Asp Gly Val Gl - #n Thr Gly Ala Val Arg# 365- CTG TAT GGC GCC AAC TCG ACG GTT TCC AGC CA - #G TCC GGC AGT GGC CAG3372Leu Tyr Gly Ala Asn Ser Thr Val Ser Ser Gl - #n Ser Gly Ser Gly Gln# 380- CAG GCG ACC CTC GAA GGC AGC GCG GGC AAC GA - #T GCG CTG AGC GGG ACC3420Gln Ala Thr Leu Glu Gly Ser Ala Gly Asn As - #p Ala Leu Ser Gly Thr# 395- GAG GCC CAC GAG ACG CTG CTC GGC CAG GCC GG - #C GAC GAC CGC CTG AAC3468Glu Ala His Glu Thr Leu Leu Gly Gln Ala Gl - #y Asp Asp Arg Leu Asn# 410- GGC GAT GCC GGC AAC GAC ATC CTC GAC GGC GG - #G GCA GGG CGC GAC AAC3516Gly Asp Ala Gly Asn Asp Ile Leu Asp Gly Gl - #y Ala Gly Arg Asp Asn415 4 - #20 4 - #25 4 -#30- CTG ACC GGC GGC GCG GGC GCC GAC ACC TTC CG - #C TTC TCC GCG CGC ACC3564Leu Thr Gly Gly Ala Gly Ala Asp Thr Phe Ar - #g Phe Ser Ala Arg Thr# 445- GAC AGC TAC CGC ACC GAC AGC GCC AGC TTC AA - #C GAC CTG ATC ACC GAC3612Asp Ser Tyr Arg Thr Asp Ser Ala Ser Phe As - #n Asp Leu Ile Thr Asp# 460- TTC GAC GCC GAC GAG GAC AGC ATC GAC CTG TC - #C GCG CTG GGC TTC ACC3660Phe Asp Ala Asp Glu Asp Ser Ile Asp Leu Se - #r Ala Leu Gly Phe Thr# 475- GGC CTG GGC GAC GGC TAC AAT GGC ACC CTG CT - #G CTG AAG ACC AAC GCC3708Gly Leu Gly Asp Gly Tyr Asn Gly Thr Leu Le - #u Leu Lys Thr Asn Ala# 490- GAG GGT ACG CGC ACC TAC CTG AAG AGC TAC GA - #A GCG GAC GCC CAG GGC3756Glu Gly Thr Arg Thr Tyr Leu Lys Ser Tyr Gl - #u Ala Asp Ala Gln Gly495 5 - #00 5 - #05 5 -#10- CGG CGC TTC GAG ATC GCC CTG GAC GGC AAC TT - #C ACC GGT CTG TTC AAC3804Arg Arg Phe Glu Ile Ala Leu Asp Gly Asn Ph - #e Thr Gly Leu Phe Asn# 525- GAC AAC AAC CTG TTG TTC GAC GCC GCT CCG GC - #C ACC GGT ACC GAG GGC3852Asp Asn Asn Leu Leu Phe Asp Ala Ala Pro Al - #a Thr Gly Thr Glu Gly# 540- AGC GAC AAC CTG CTC GGC ACC GAC GCC GGG GA - #A ACC CTC CTG GGC TAC3900Ser Asp Asn Leu Leu Gly Thr Asp Ala Gly Gl - #u Thr Leu Leu Gly Tyr# 555- GGC GGC AAC GAC ACC CTC AAC GGC GGG GCC GG - #C GAC GAC ATC CTG GTC3948Gly Gly Asn Asp Thr Leu Asn Gly Gly Ala Gl - #y Asp Asp Ile Leu Val# 570- GGC GGC GCC GGG CGC GAC AGC CTG ACC GGC GG - #C GCC GGG GCG GAC GTG3996Gly Gly Ala Gly Arg Asp Ser Leu Thr Gly Gl - #y Ala Gly Ala Asp Val575 5 - #80 5 - #85 5 -#90- TTC CGC TTC GAC GCG CTG TCC GAC AGC CAG CG - #C AAC TAC ACC ACC GGC4044Phe Arg Phe Asp Ala Leu Ser Asp Ser Gln Ar - #g Asn Tyr Thr Thr Gly# 605- GAC AAC CAG GCC GAC CGC ATT CTC GAC TTC GA - #C CCG ACC CTG GAC AGG4092Asp Asn Gln Ala Asp Arg Ile Leu Asp Phe As - #p Pro Thr Leu Asp Arg# 620- ATC GAC GTG TCG GCG CTG GGC TTC ACC GGG CT - #G GGC AAC GGC CGC AAC4140Ile Asp Val Ser Ala Leu Gly Phe Thr Gly Le - #u Gly Asn Gly Arg Asn# 635- GGC ACC CTC GCC GTG GTG CTC AAC AGC GCC GG - #C GAC CGC ACC GAT CTG4188Gly Thr Leu Ala Val Val Leu Asn Ser Ala Gl - #y Asp Arg Thr Asp Leu# 650- AAG AGC TAC GAC ACC GAC GCC AAC GGC TAC AG - #C TTC GAG CTT TCC CTC4236Lys Ser Tyr Asp Thr Asp Ala Asn Gly Tyr Se - #r Phe Glu Leu Ser Leu655 6 - #60 6 - #65 6 -#70- GCG GGC AAC TAC CAG GGG CAG CTC AGC GCC GA - #G CAG TTC GTT TTC GCG4284Ala Gly Asn Tyr Gln Gly Gln Leu Ser Ala Gl - #u Gln Phe Val Phe Ala# 685- ACG TCT CAG GGG GGA CAG ATG ACG ATT ATC GA - #A GGC ACC GAC GGC AAC4332Thr Ser Gln Gly Gly Gln Met Thr Ile Ile Gl - #u Gly Thr Asp Gly Asn# 700- GAT ACC TTG CAG GGC ACC GAG GCC AAC GAG CG - #G CTC CTC GGC CTG GAC4380Asp Thr Leu Gln Gly Thr Glu Ala Asn Glu Ar - #g Leu Leu Gly Leu Asp# 715- GGC CGG GAC AAC CTG AAC GGC GGC GCC GGC GA - #C GAC ATC CTC GAC GGC4428Gly Arg Asp Asn Leu Asn Gly Gly Ala Gly As - #p Asp Ile Leu Asp Gly# 730- GGA GCG GGG CGC GAC ACC CTG ACC GGC GGC AC - #G GGG GCC GAC ACC TTC4476Gly Ala Gly Arg Asp Thr Leu Thr Gly Gly Th - #r Gly Ala Asp Thr Phe735 7 - #40 7 - #45 7 -#50- CTG TTC TCC ACG CGT ACC GAC AGC TAC CGC AC - #C GAC AGC GCC AGC TTC4524Leu Phe Ser Thr Arg Thr Asp Ser Tyr Arg Th - #r Asp Ser Ala Ser Phe# 765- AAC GAC CTG ATC ACC GAC TTC GAT CCC ACC CA - #G GAC CGC ATC GAC CTG4572Asn Asp Leu Ile Thr Asp Phe Asp Pro Thr Gl - #n Asp Arg Ile Asp Leu# 780- TCC GGC CTG GGC TTC AGC GGT TTC GGC AAC GG - #C TAC GAC GGC ACC CTG4620Ser Gly Leu Gly Phe Ser Gly Phe Gly Asn Gl - #y Tyr Asp Gly Thr Leu# 795- CTG CTG CAG GTC AAC GCC GCG GGC ACC CGC AC - #C TAC CTG AAG AGT TTC4668Leu Leu Gln Val Asn Ala Ala Gly Thr Arg Th - #r Tyr Leu Lys Ser Phe# 810- GAG GCC GAT GCC AAC GGC CAG CGC TTC GAG AT - #C GCC CTG GAC GGC GAC4716Glu Ala Asp Ala Asn Gly Gln Arg Phe Glu Il - #e Ala Leu Asp Gly Asp815 8 - #20 8 - #25 8 -#30- TTC AGC GGC CAA TTG GAC AGC GGC AAC GTG AT - #C TTC GAG CCC GCC GTG4764Phe Ser Gly Gln Leu Asp Ser Gly Asn Val Il - #e Phe Glu Pro Ala Val# 845- TTC AAT GCC AAG GAC TTC GGC GCG CTG GGC GA - #C GGC GCC AGC GAC GAC4812Phe Asn Ala Lys Asp Phe Gly Ala Leu Gly As - #p Gly Ala Ser Asp Asp# 860- CGG CCG GCC ATC CAG GCG GCG ATC GAC GCC GC - #C TAC GCG GCC GGT GGC4860Arg Pro Ala Ile Gln Ala Ala Ile Asp Ala Al - #a Tyr Ala Ala Gly Gly# 875- GGC ACC GTC TAC CTG CCG GCC GGC GAG TAC CG - #G GTC AGC CCC ACC GGG4908Gly Thr Val Tyr Leu Pro Ala Gly Glu Tyr Ar - #g Val Ser Pro Thr Gly# 890- GAG CCG GGC GAC GGC TGC CTG ATG CTC AAG GA - #C GGC GTC TAC CTG GCC4956Glu Pro Gly Asp Gly Cys Leu Met Leu Lys As - #p Gly Val Tyr Leu Ala895 9 - #00 9 - #05 9 -#10- GGC GAC GGC ATA GGC GAA ACG GTC ATC AAG CT - #G ATC GAC GGC TCC GAC5004Gly Asp Gly Ile Gly Glu Thr Val Ile Lys Le - #u Ile Asp Gly Ser Asp# 925- CAG AAG ATC ACC GGC ATG GTG CGC TCG GCC TA - #T GGC GAA GAG ACC AGC5052Gln Lys Ile Thr Gly Met Val Arg Ser Ala Ty - #r Gly Glu Glu Thr Ser# 940- AAC TTC GGC ATG AGC GAC CTG ACC CTC GAC GG - #C AAC CGC GAC AAC ACC5100Asn Phe Gly Met Ser Asp Leu Thr Leu Asp Gl - #y Asn Arg Asp Asn Thr# 955- AGC GGC AAG GTC GAC GGC TGG TTC AAC GGC TA - #C ATC CCC GGC CAG GAC5148Ser Gly Lys Val Asp Gly Trp Phe Asn Gly Ty - #r Ile Pro Gly Gln Asp# 970- GGC GCC GAC CGC AAC GTG ACC ATC GAG CGG GT - #G GAA ATC CGC GAG ATG5196Gly Ala Asp Arg Asn Val Thr Ile Glu Arg Va - #l Glu Ile Arg Glu Met975 9 - #80 9 - #85 9 -#90- TCC GGC TAT GGC TTC GAT CCG CAC GAG CAG AC - #C ATC AAC CTG ACG ATC5244Ser Gly Tyr Gly Phe Asp Pro His Glu Gln Th - #r Ile Asn Leu Thr Ile# 10050- CGC GAC AGC GTG GCC CAC GAC AAC GGC CTC GA - #C GGC TTC GTC GCC GAC5292Arg Asp Ser Val Ala His Asp Asn Gly Leu As - #p Gly Phe Val Ala Asp# 10205- TAC CTG GTC GAC AGC GTG TTC GAG AAC AAC GT - #C GCC TAC AAC AAC GAC5340Tyr Leu Val Asp Ser Val Phe Glu Asn Asn Va - #l Ala Tyr Asn Asn Asp# 10350- CGC CAC GGC TTC AAC ATC GTC ACC AGC ACC TA - #C GAT TTC GTC ATG ACC5388Arg His Gly Phe Asn Ile Val Thr Ser Thr Ty - #r Asp Phe Val Met Thr# 10505- AAC AAC GTC GCC TAC GGC AAC GGC GGC GCC GG - #C CTG ACG ATC CAG CGG5436Asn Asn Val Ala Tyr Gly Asn Gly Gly Ala Gl - #y Leu Thr Ile Gln Arg# 10701060 - # 1065- GGC TCG GAG GAC CTG GCC CAG CCG ACC GAT AT - #C CTG ATC GAC GGC GGC5484Gly Ser Glu Asp Leu Ala Gln Pro Thr Asp Il - #e Leu Ile Asp Gly Gly# 10850- GCC TAC TAC GAC AAC GCC CTG GAA GGC GTG CT - #G TTC AAG ATG ACC AAC5532Ala Tyr Tyr Asp Asn Ala Leu Glu Gly Val Le - #u Phe Lys Met Thr Asn# 11005- AAC GTC ACC CTG CAG AAC GCC GAG ATC TAC GG - #C AAC GGC TCC TCC GGC5580Asn Val Thr Leu Gln Asn Ala Glu Ile Tyr Gl - #y Asn Gly Ser Ser Gly# 11150- GTG CGC CTG TAC GGC ACG GAG GAC GTG CAG AT - #C CTC GAC AAC CAG ATC5628Val Arg Leu Tyr Gly Thr Glu Asp Val Gln Il - #e Leu Asp Asn Gln Ile# 11305- CAC GAC AAT TCG CAG AAC GGC ACC TAT CCG GA - #A GTC CTG CTG CAG GCC5676His Asp Asn Ser Gln Asn Gly Thr Tyr Pro Gl - #u Val Leu Leu Gln Ala# 11501140 - # 1145- TTC GAC GAC AGC CAG GTC ACC GGT GAG CTG TA - #C GAG ACC CTG AAC ACC5724Phe Asp Asp Ser Gln Val Thr Gly Glu Leu Ty - #r Glu Thr Leu Asn Thr# 11650- CGG ATC GAA GGC AAT CTC ATC GAC GCT TCG GA - #C AAC GCC AAC TAT GCG5772Arg Ile Glu Gly Asn Leu Ile Asp Ala Ser As - #p Asn Ala Asn Tyr Ala# 11805- GTG CGC GAG CGC GAC GAC GGC AGC GAC TAC AC - #C ACG CTC GTG GAC AAC5820Val Arg Glu Arg Asp Asp Gly Ser Asp Tyr Th - #r Thr Leu Val Asp Asn# 11950- GAC ATC AGC GGC GGC CAG GTC GCC TCG GTG CA - #G CTT TCC GGC GCC CAT5868Asp Ile Ser Gly Gly Gln Val Ala Ser Val Gl - #n Leu Ser Gly Ala His# 12105- TCG AGT CTT TCC GGC GGC ACC GTC GAA GTG CC - #G CAG GGG ACC GAC GGC5916Ser Ser Leu Ser Gly Gly Thr Val Glu Val Pr - #o Gln Gly Thr Asp Gly# 12301220 - # 1225- AAC GAC GTG CTG GTC GGC AGC GAT GCC AAC GA - #C CAG CTC TAC GGC GGA5964Asn Asp Val Leu Val Gly Ser Asp Ala Asn As - #p Gln Leu Tyr Gly Gly# 12450- GCC GGC GAC GAC CGC CTG GAC GGC GGC GCC GG - #T GAC GAC CTG CTC GAC6012Ala Gly Asp Asp Arg Leu Asp Gly Gly Ala Gl - #y Asp Asp Leu Leu Asp# 12605- GGC GGA GCG GGG CGC GAC GAC CTG ACC GGC GG - #C ACG GGT GCC GAC ACC6060Gly Gly Ala Gly Arg Asp Asp Leu Thr Gly Gl - #y Thr Gly Ala Asp Thr# 12750- TTC GTG TTC GCC GCG CGT ACC GAT AGC TAC CG - #C ACC GAC GCG GGG GTG6108Phe Val Phe Ala Ala Arg Thr Asp Ser Tyr Ar - #g Thr Asp Ala Gly Val# 12905- TTC AAC GAC CTG ATC CTC GAC TTC GAC GCC AG - #C GAG GAC CGC ATC GAC6156Phe Asn Asp Leu Ile Leu Asp Phe Asp Ala Se - #r Glu Asp Arg Ile Asp# 13101300 - # 1305- CTG TCC GCC CTG GGT TTC AGC GGC TTC GGC GA - #C GGC TAC AAC GGC ACC6204Leu Ser Ala Leu Gly Phe Ser Gly Phe Gly As - #p Gly Tyr Asn Gly Thr# 13250- CTG CTG GTG CAG CTC AGC AGC GCC GGA ACC CG - #T ACC TAC CTC AAG AGC6252Leu Leu Val Gln Leu Ser Ser Ala Gly Thr Ar - #g Thr Tyr Leu Lys Ser# 13405- TAC GAG GAG GAC CTC GAG GGC CGG CGC TTC GA - #G GTC GCC CTG GAC GGC6300Tyr Glu Glu Asp Leu Glu Gly Arg Arg Phe Gl - #u Val Ala Leu Asp Gly# 13550- GAC CAC ACG GGC GAT CTT TCC GCC GCC AAT GT - #G GTT TTC GCC GAC GAC6348Asp His Thr Gly Asp Leu Ser Ala Ala Asn Va - #l Val Phe Ala Asp Asp# 13705- GGC TCG GCC GCC GTG GCG AGC AGC GAT CCC GC - #C GCC ACA CAG TTG GAG6396Gly Ser Ala Ala Val Ala Ser Ser Asp Pro Al - #a Ala Thr Gln Leu Glu# 13901380 - # 1385- GTG GTC GGC AGC AGC GGC ACC CAG ACC GAT CA - #A CTC GCC TGA#6438Val Val Gly Ser Ser Gly Thr Gln Thr Asp Gl - #n Leu Ala *# 1400- TCCGACCCCG CCCATACCCG CCCGGCCATT CCGGCCGGGC GAACCAATGG TC - #TTCAGGCC6498- AGTCTCAGGC ACAGCAGCGC GCGAGCCGCT TCGCTTTGTC CGCCCCCCGC TT - #TTCTCGCT6558- GAACGCGACG ATCGCCGGGC GCCGGGGAAG GGTTCGCCGC ATGCCGAGCC GG - #GGACGGGA6618- AAAGCCTGTT CGACCAGTCG ACTCTTCCTC CCTTCACTTT CCAGGCAGCC TG - #CGGGCTGC6678- GCAGTAACGG AACAGGAAGC AGC ATG GAT TAC AAC GTC AA - #A GAT TTC GGG6728#Asp Phe Gly Asp Tyr Asn Val Lys# 5 1- GCG CTG GGC GAT GGC GTC AGC GAC GAT ACG GC - #C GCC ATC CAG GCG GCG6776Ala Leu Gly Asp Gly Val Ser Asp Asp Thr Al - #a Ala Ile Gln Ala Ala# 25- ATC GAT GCC GCC TAC GCG GCC GGC GGC GGC AC - #C GTC TAC CTG CCG GCC6824Ile Asp Ala Ala Tyr Ala Ala Gly Gly Gly Th - #r Val Tyr Leu Pro Ala# 40- GGC GAA TAC CGG GTC AGC GGC GGC GAG GAG CC - #T TCC GAT GGT TGC CTG6872Gly Glu Tyr Arg Val Ser Gly Gly Glu Glu Pr - #o Ser Asp Gly Cys Leu# 55- ACC ATC AAG AGC AAC GTC CAT ATC GTC GGC GC - #G GGG ATG GGC GAG ACG6920Thr Ile Lys Ser Asn Val His Ile Val Gly Al - #a Gly Met Gly Glu Thr# 70- GTC ATC AAG CTG GTC GAC GGC TGG GAT CAG GA - #C GTC ACC GGC ATC GTC6968Val Ile Lys Leu Val Asp Gly Trp Asp Gln As - #p Val Thr Gly Ile Val# 85- CGC TCG GCC TAC GGC GAG GAG ACC AGC AAC TT - #C GGC ATG AGC GAC CTG7016Arg Ser Ala Tyr Gly Glu Glu Thr Ser Asn Ph - #e Gly Met Ser Asp Leu#105- ACC CTC GAC GGC AAC CGC GAC AAC ACC AGC GG - #C AAG GTC GAC GGC TGG7064Thr Leu Asp Gly Asn Arg Asp Asn Thr Ser Gl - #y Lys Val Asp Gly Trp# 120- TTC AAC GGC TAC ATT CCC GGC GAG GAC GGC GC - #C GAC CGC GAC GTG ACC7112Phe Asn Gly Tyr Ile Pro Gly Glu Asp Gly Al - #a Asp Arg Asp Val Thr# 135- CTG GAG CGG GTG GAA ATC CGT GAA ATG TCC GG - #T TAC GGT TTC GAT CCG7160Leu Glu Arg Val Glu Ile Arg Glu Met Ser Gl - #y Tyr Gly Phe Asp Pro# 150- CAC GAG CAG ACC ATC AAC CTG ACG ATC CGC GA - #C AGC GTG GCC CAC GAC7208His Glu Gln Thr Ile Asn Leu Thr Ile Arg As - #p Ser Val Ala His Asp# 165- AAC GGC CTC GAC GGC TTC GTC GCC GAT TTC CA - #G ATC GGC GGG GTG TTC7256Asn Gly Leu Asp Gly Phe Val Ala Asp Phe Gl - #n Ile Gly Gly Val Phe170 1 - #75 1 - #80 1 -#85- GAG AAC AAC GTC TCG TAC AAC AAC GAC CGC CA - #C GGC TTC AAC ATC GTC7304Glu Asn Asn Val Ser Tyr Asn Asn Asp Arg Hi - #s Gly Phe Asn Ile Val# 200- ACC AGC ACC AAC GAC TTC GTC CTG AGC AAC AA - #C GTC GCC TAC GGC AAC7352Thr Ser Thr Asn Asp Phe Val Leu Ser Asn As - #n Val Ala Tyr Gly Asn# 215- GGC GGC GCC GGC CTG GTG GTG CAG CGC GGC TC - #G TCC GAC GTG GCG CAC7400Gly Gly Ala Gly Leu Val Val Gln Arg Gly Se - #r Ser Asp Val Ala His# 230- CCC TAC GAC ATC CTG ATC GAC GGC GGC GCC TA - #C TAC GAC AAC GGC CTG7448Pro Tyr Asp Ile Leu Ile Asp Gly Gly Ala Ty - #r Tyr Asp Asn Gly Leu# 245- GAA GGC GTG CAG ATC AAG ATG GCC CAC GAC GT - #C ACC CTG CAG AAC GCC7496Glu Gly Val Gln Ile Lys Met Ala His Asp Va - #l Thr Leu Gln Asn Ala250 2 - #55 2 - #60 2 -#65- GAG ATC TAC GGC AAC GGC CTA TAC GGG GTG CG - #C GTC TAC GGC GCC GAG7544Glu Ile Tyr Gly Asn Gly Leu Tyr Gly Val Ar - #g Val Tyr Gly Ala Glu# 280- GAT GTG CAG ATC CTC GAC AAC TAC ATC CAC GA - #C AAT TCG CAG AAC GGT7592Asp Val Gln Ile Leu Asp Asn Tyr Ile His As - #p Asn Ser Gln Asn Gly# 295- TCC TAC GCG GAA ATC CTC CTG CAG TCC TAC GA - #C GAT ACC GCC GGG GTG7640Ser Tyr Ala Glu Ile Leu Leu Gln Ser Tyr As - #p Asp Thr Ala Gly Val# 310- TCC GGC AAT TTC TAC ACC ACC ACC GGC ACC TG - #G ATC GAA GGC AAC ACC7688Ser Gly Asn Phe Tyr Thr Thr Thr Gly Thr Tr - #p Ile Glu Gly Asn Thr# 325- ATC GTC GGC TCG GCC AAC TCC ACC TAT GGC AT - #C CAG GAG CGC GAC GAC7736Ile Val Gly Ser Ala Asn Ser Thr Tyr Gly Il - #e Gln Glu Arg Asp Asp330 3 - #35 3 - #40 3 -#45- GGC ACC GAC TAC AGC AGC CTC TAC GCC AAC AG - #C GTC AGC AAT GTG CAG7784Gly Thr Asp Tyr Ser Ser Leu Tyr Ala Asn Se - #r Val Ser Asn Val Gln# 360- AAC GGC TCG GTG CGC CTC TAC GGC GCC AAC TC - #C GTC GTC TCC GAC CTG7832Asn Gly Ser Val Arg Leu Tyr Gly Ala Asn Se - #r Val Val Ser Asp Leu# 375- CCC GGC ACC GGC CAG CAG GCG ACC CTC GAA GG - #C ACG GCC GGC AAC GAC7880Pro Gly Thr Gly Gln Gln Ala Thr Leu Glu Gl - #y Thr Ala Gly Asn Asp# 390- ACG CTT GGC GGC AGC GAC GCC CAC GAG ACG CT - #G CTC GGG CTG GAC GGC7928Thr Leu Gly Gly Ser Asp Ala His Glu Thr Le - #u Leu Gly Leu Asp Gly# 405- AAC GAC CGC CTG AAC GGC GGC GCC GGC AAC GA - #C ATC CTC GAC GGC GGC7976Asn Asp Arg Leu Asn Gly Gly Ala Gly Asn As - #p Ile Leu Asp Gly Gly410 4 - #15 4 - #20 4 -#25- GCC GGG CGC GAC AAC CTG ACC GGC GGC GCG GG - #C GCC GAC CTG TTC CGC8024Ala Gly Arg Asp Asn Leu Thr Gly Gly Ala Gl - #y Ala Asp Leu Phe Arg# 440- GTC TCC GCG CGC ACC GAC AGC TAC CGC ACC GA - #C AGC GCC AGC TTC AAC8072Val Ser Ala Arg Thr Asp Ser Tyr Arg Thr As - #p Ser Ala Ser Phe Asn# 455- GAC CTG ATC ACC GAC TTC GAC GCC AGC CAG GA - #C CGC ATC GAC CTG TCC8120Asp Leu Ile Thr Asp Phe Asp Ala Ser Gln As - #p Arg Ile Asp Leu Ser# 470- GCG CTG GGC TTC ACC GGG CTG GGC GAC GGC TA - #T AAC GGC ACC CTG CTG8168Ala Leu Gly Phe Thr Gly Leu Gly Asp Gly Ty - #r Asn Gly Thr Leu Leu# 485- CTG CAG GTC AGC GCC GAC GGC AGC CGC ACC TA - #T CTG AAG AGC CTG GAG8216Leu Gln Val Ser Ala Asp Gly Ser Arg Thr Ty - #r Leu Lys Ser Leu Glu490 4 - #95 5 - #00 5 -#05- GCG GAT GCC GAG GGG CGG CGT TTC GAG ATC GC - #C CTG GAC GGC AAC TTC8264Ala Asp Ala Glu Gly Arg Arg Phe Glu Ile Al - #a Leu Asp Gly Asn Phe# 520- GCC GGC CTG CTC GGT GCC GGC AAC CTG CTC TT - #C GAG CGC ACC GCC ATC8312Ala Gly Leu Leu Gly Ala Gly Asn Leu Leu Ph - #e Glu Arg Thr Ala Ile# 535- GAG GGG GAT GCC GGC GAC AAC GCC CTG CTC GG - #T ACC TCG GCC GCC GAG8360Glu Gly Asp Ala Gly Asp Asn Ala Leu Leu Gl - #y Thr Ser Ala Ala Glu# 550- ACA TTG CTC GGC CAC GCC GGC AAC GAC ACG CT - #C GAC GGC GGG GCC GGC8408Thr Leu Leu Gly His Ala Gly Asn Asp Thr Le - #u Asp Gly Gly Ala Gly# 565- GAC GAC ATC CTG GTC GGC GGC GCC GGG CGC GA - #C AGC CTC ACC GGC GGC8456Asp Asp Ile Leu Val Gly Gly Ala Gly Arg As - #p Ser Leu Thr Gly Gly570 5 - #75 5 - #80 5 -#85- GCC GGA GCG GAC GTG TTC CGC TTC GAC GCG CT - #G TCC GAC AGC CAG CGC8504Ala Gly Ala Asp Val Phe Arg Phe Asp Ala Le - #u Ser Asp Ser Gln Arg# 600- AAC TAC GAC ATC GGC GAC AAC CAG GGC GAC CG - #C ATC GCC GAC TTC GCG8552Asn Tyr Asp Ile Gly Asp Asn Gln Gly Asp Ar - #g Ile Ala Asp Phe Ala# 615- GTG GGC GAA GAC AAG CTC GAC GTA TCG GCG CT - #G GGC TTC ACC GGG CTG8600Val Gly Glu Asp Lys Leu Asp Val Ser Ala Le - #u Gly Phe Thr Gly Leu# 630- GGC GAC GGC TAC AAC GGC ACC CTC GCC CTG GT - #G CTC AAC AGC GCC GGC8648Gly Asp Gly Tyr Asn Gly Thr Leu Ala Leu Va - #l Leu Asn Ser Ala Gly# 645- GAC CGC ACC TAC GTG AAA AGC TAC GAG AAC GG - #C GCC GAC GGC TAC CGC8696Asp Arg Thr Tyr Val Lys Ser Tyr Glu Asn Gl - #y Ala Asp Gly Tyr Arg650 6 - #55 6 - #60 6 -#65- TTC GAG TTT TCC CTC GAC GGC AAC TAT CTG GA - #G CTA CTC GGC AAC GAG8744Phe Glu Phe Ser Leu Asp Gly Asn Tyr Leu Gl - #u Leu Leu Gly Asn Glu# 680- GAT TTC ATC TTC GCC ACG CCC AGC GGC CAG CA - #A CTC CTC GAA GGC AGC8792Asp Phe Ile Phe Ala Thr Pro Ser Gly Gln Gl - #n Leu Leu Glu Gly Ser# 695- GCC GGC AAC GAC AGC CTG CAG GGC ACG GCC GC - #C GAC GAG GTG ATC CAC8840Ala Gly Asn Asp Ser Leu Gln Gly Thr Ala Al - #a Asp Glu Val Ile His# 710- GGC GGC GGC GGG CGC GAC ACG CTG GCC GGA GG - #G GCC GGG GCC GAC GTG8888Gly Gly Gly Gly Arg Asp Thr Leu Ala Gly Gl - #y Ala Gly Ala Asp Val# 725- TTC CGC TTT AGC GAA CTG ACC GAC AGC TAC CG - #A GAC AGT GCC AGC TAT8936Phe Arg Phe Ser Glu Leu Thr Asp Ser Tyr Ar - #g Asp Ser Ala Ser Tyr730 7 - #35 7 - #40 7 -#45- GCC GAT CTG ATC ACT GAC TTC GAT GCC AGC GA - #G GAT CGT ATC GAC CTG8984Ala Asp Leu Ile Thr Asp Phe Asp Ala Ser Gl - #u Asp Arg Ile Asp Leu# 760- TCC GGC CTC GGC TTC AGC GGT CTG GGC AAC GG - #C TAC GGC GGT ACC CTG9032Ser Gly Leu Gly Phe Ser Gly Leu Gly Asn Gl - #y Tyr Gly Gly Thr Leu# 775- GCG CTG CAG GTG AAC AGC GCC GGT ACC CGC AC - #C TAC CTG AAG AGC TTC9080Ala Leu Gln Val Asn Ser Ala Gly Thr Arg Th - #r Tyr Leu Lys Ser Phe# 790- GAG ACC AAC GCC GCC GGC GAG CGT TTC GAG AT - #C GCC CTG GAC GGC GAC9128Glu Thr Asn Ala Ala Gly Glu Arg Phe Glu Il - #e Ala Leu Asp Gly Asp# 805- CTG TCC GCG CTC GGC GGG GCC AAC CTG ATC CT - #C GAC GCG CGT ACC GTA9176Leu Ser Ala Leu Gly Gly Ala Asn Leu Ile Le - #u Asp Ala Arg Thr Val810 8 - #15 8 - #20 8 -#25- CTG GCG GGC GGC GAC GGC AAC GAC ACG CTT TC - #C GGC AGC AGC GCG GCC9224Leu Ala Gly Gly Asp Gly Asn Asp Thr Leu Se - #r Gly Ser Ser Ala Ala# 840- GAG GAA CTG CTC GGC GGG GTC GGC AAC GAC AG - #C CTG GAC GGC GGC GCC9272Glu Glu Leu Leu Gly Gly Val Gly Asn Asp Se - #r Leu Asp Gly Gly Ala# 855- GGC AAC GAC ATC CTC GAC GGC GGG GCG GGG CG - #C GAC ACC CTG AGT GGC9320Gly Asn Asp Ile Leu Asp Gly Gly Ala Gly Ar - #g Asp Thr Leu Ser Gly# 870- GGC AGC GGC AGC GAC ATC TTC CGC TTC GGC GG - #C GCG CTC GAC AGC TTC9368Gly Ser Gly Ser Asp Ile Phe Arg Phe Gly Gl - #y Ala Leu Asp Ser Phe# 885- CGC AAC TAC GCC AGC GGG ACG AAC GGC ACC GA - #C AGC ATC ACC GAC TTC9416Arg Asn Tyr Ala Ser Gly Thr Asn Gly Thr As - #p Ser Ile Thr Asp Phe890 8 - #95 9 - #00 9 -#05- ACC CCC GGC GAG GAT CTG ATC GAC CTC TCC GT - #G CTC GGC TAC ACC GGG9464Thr Pro Gly Glu Asp Leu Ile Asp Leu Ser Va - #l Leu Gly Tyr Thr Gly# 920- CTG GGC GAC GGC TAC AAC GGT ACC CTG GCG AT - #A GTG CTG AAC GAC GCC9512Leu Gly Asp Gly Tyr Asn Gly Thr Leu Ala Il - #e Val Leu Asn Asp Ala# 935- GGC ACC AAG ACC TAC CTG AAA AAC CGC GAG AG - #C GAC GCC GAA GGC AAC9560Gly Thr Lys Thr Tyr Leu Lys Asn Arg Glu Se - #r Asp Ala Glu Gly Asn# 950- CAG TTC GAG ATC GCC CTG GAG GGC AAC CAC GC - #C GAC CAG CTC GAT GCG9608Gln Phe Glu Ile Ala Leu Glu Gly Asn His Al - #a Asp Gln Leu Asp Ala# 965- AGC GAC TTC ATC TTC GCC ACG GCG GCC GCG AC - #C ACC GGA ATC GAG GTG9656Ser Asp Phe Ile Phe Ala Thr Ala Ala Ala Th - #r Thr Gly Ile Glu Val970 9 - #75 9 - #80 9 -#85- GTC GGC GGC AGC GGC ACC CAG ACC GAT CAG CT - #C GCC TGA TCCGACCCCG9705Val Gly Gly Ser Gly Thr Gln Thr Asp Gln Le - #u Ala *# 995- CCCGCACCCG CCCGGCCATT CCGGCCGGGC GAACCAATGG CCTTTTGATC AG - #TCTCAGGC9765- ACAGCAACGT GTGCGCCGCT TCGCTTGTTC GCCCTCCCGG CCTTGTTTCT CG - #CTGAAAGC9825- GACGATCGCC GGGGGCGTGC CGGGCGCGAG AAAAGGTTCG CCGTGTGCAA AG - #CCGGGGAC9885- GGGAAAAGCC TGTTCAAGTA GTCGACTCTT CCTTCTCCTT TTTCCTAGAC GG - #CCTCTTGG9945#AAA GAT TTC 9996 AAGCAGC ATG GAC TTC AAC GTC# Met Asp Phe As - #n Val Lys Asp Phe# 5 1- GGG GCA CTG GGC GAT GGC GCC AGC GAC GAC AC - #G GCG GCC ATC CAG GCG10044Gly Ala Leu Gly Asp Gly Ala Ser Asp Asp Th - #r Ala Ala Ile Gln Ala# 20- GCG ATC GAT GCC GCC CAC GCG GCG GGC GGC GG - #C ACC GTC TAC CTG CCG10092Ala Ile Asp Ala Ala His Ala Ala Gly Gly Gl - #y Thr Val Tyr Leu Pro# 40- GCT GGC GAG TAT CGG GTC AGC GGC GGC GAG GA - #G CCT TCC GAC GGC GCG10140Ala Gly Glu Tyr Arg Val Ser Gly Gly Glu Gl - #u Pro Ser Asp Gly Ala# 55- CTG ACC ATC AAG AGC AAC GTC TAT ATC GTC GG - #C GCC GGG ATG GGC GAG10188Leu Thr Ile Lys Ser Asn Val Tyr Ile Val Gl - #y Ala Gly Met Gly Glu# 70- ACG GTG ATC AAG ATG GTC GAC GGC TGG ACG CA - #G AAC GTC ACC GGC ATG10236Thr Val Ile Lys Met Val Asp Gly Trp Thr Gl - #n Asn Val Thr Gly Met# 85- GTG CGC TCG GCC TAT GGC GAG GAG ACC AGC AA - #C TTC GGC ATG AGC GAC10284Val Arg Ser Ala Tyr Gly Glu Glu Thr Ser As - #n Phe Gly Met Ser Asp# 100- CTG ACC CTC GAC GGC AAC CGC GAC AAC CTG TC - #C GCC AAG GTC GAC GGC10332Leu Thr Leu Asp Gly Asn Arg Asp Asn Leu Se - #r Ala Lys Val Asp Gly105 1 - #10 1 - #15 1 -#20- TGG TTC AAC GGC TAC ATT CCC GGC CAG GAC GG - #T GCC GAT CGC GAC GTG10380Trp Phe Asn Gly Tyr Ile Pro Gly Gln Asp Gl - #y Ala Asp Arg Asp Val# 135- ACC CTG GAG CGG GTG GAA ATC CGC GAA ATG TC - #C GGT TAC GGT TTC GAT10428Thr Leu Glu Arg Val Glu Ile Arg Glu Met Se - #r Gly Tyr Gly Phe Asp# 150- CCG CAC GAG CAG ACC ATC AAC CTG ACG ATC CG - #C GAC AGC GTG GCC CAC10476Pro His Glu Gln Thr Ile Asn Leu Thr Ile Ar - #g Asp Ser Val Ala His# 165- GAC AAC GGC CTC GAC GGC TTC GTC GCC GAC TA - #C CAG GTC GGC GGG GTG10524Asp Asn Gly Leu Asp Gly Phe Val Ala Asp Ty - #r Gln Val Gly Gly Val# 180- TTC GAG AAC AAC GTC TCG TAC AAC AAC GAC CG - #C CAC GGC TTC AAC ATC10572Phe Glu Asn Asn Val Ser Tyr Asn Asn Asp Ar - #g His Gly Phe Asn Ile185 1 - #90 1 - #95 2 -#00- GTC ACC AGC ACC AAC GAC TTC GTC CTG AGC AA - #C AAC GTC GCC TAC GGC10620Val Thr Ser Thr Asn Asp Phe Val Leu Ser As - #n Asn Val Ala Tyr Gly# 215- AAC GGC GGC GCC GGC CTG GTG GTG CAG CGC GG - #C TCG TAC GAC CTG CCC10668Asn Gly Gly Ala Gly Leu Val Val Gln Arg Gl - #y Ser Tyr Asp Leu Pro# 230- CAT CCC TAC GAC ATC CTG ATC GAC GGC GGC GC - #C TAC TAC GAC AAC GCC10716His Pro Tyr Asp Ile Leu Ile Asp Gly Gly Al - #a Tyr Tyr Asp Asn Ala# 245- TTG GAA GGC GTG CAG CTC AAG ATG ACC CAC GA - #C GTC ACC CTG CAG AAC10764Leu Glu Gly Val Gln Leu Lys Met Thr His As - #p Val Thr Leu Gln Asn# 260- GCC GAG ATC TAT GGC AAC GGC CTG TAC GGG GT - #G CGC GTC TAC GGC GCC10812Ala Glu Ile Tyr Gly Asn Gly Leu Tyr Gly Va - #l Arg Val Tyr Gly Ala265 2 - #70 2 - #75 2 -#80- CAG GAC GTG CAA CTC CTC GAT AAC CAG ATC CA - #C GAC AAT TCG CAG AAC10860Gln Asp Val Gln Leu Leu Asp Asn Gln Ile Hi - #s Asp Asn Ser Gln Asn# 295- GGC GCC TAT GCC GAA GTC CTG CTG CAG TCC TA - #C GAC GAC ACC GCC GGG10908Gly Ala Tyr Ala Glu Val Leu Leu Gln Ser Ty - #r Asp Asp Thr Ala Gly# 310- GTG TCC GGC AAC TTT TAC GTC ACC ACC GGC AC - #C TGG CTC GAA GGC AAC10956Val Ser Gly Asn Phe Tyr Val Thr Thr Gly Th - #r Trp Leu Glu Gly Asn# 325- GTC ATC AGC GGC TCG GCC AAT TCC ACC TTC GG - #C ATC CAG GAG CGC GCC11004Val Ile Ser Gly Ser Ala Asn Ser Thr Phe Gl - #y Ile Gln Glu Arg Ala# 340- GAC GGC ACC GAC TAC AGC AGC CTT TAC GCC AA - #T ACC ATC GAC GGC GTG11052Asp Gly Thr Asp Tyr Ser Ser Leu Tyr Ala As - #n Thr Ile Asp Gly Val345 3 - #50 3 - #55 3 -#60- CAG AAC GGG ACG GTA CGG CTG TAT GGC GCC AA - #C TCC ACG GTT TCC GAG11100Gln Asn Gly Thr Val Arg Leu Tyr Gly Ala As - #n Ser Thr Val Ser Glu# 375- CAG CCC AGC AGC GGC CAG CAG GCG ACC CTC GA - #A GGC ACC GCG GGC AAC11148Gln Pro Ser Ser Gly Gln Gln Ala Thr Leu Gl - #u Gly Thr Ala Gly Asn# 390- GAC GTG CTC AGC GGA ACG GGT GCC CAC GAG CT - #G ATT CTC GGC CTG GCC11196Asp Val Leu Ser Gly Thr Gly Ala His Glu Le - #u Ile Leu Gly Leu Ala# 405- GGC AAC GAT CGC CTG GAC GGT GGC GCC GGC GA - #C GAC ACC CTC GAC GGC11244Gly Asn Asp Arg Leu Asp Gly Gly Ala Gly As - #p Asp Thr Leu Asp Gly# 420- GGC GCG GGG CGC GAT ACC CTG ACC GGC GGC GC - #G GGC GCC GAT ACC TTC11292Gly Ala Gly Arg Asp Thr Leu Thr Gly Gly Al - #a Gly Ala Asp Thr Phe425 4 - #30 4 - #35 4 -#40- CGC TTC TCT GCC CGC GAG GAC AGT CAC CGC AC - #C GAC AGC GCC AGC TTC11340Arg Phe Ser Ala Arg Glu Asp Ser His Arg Th - #r Asp Ser Ala Ser Phe# 455- ACC GAC CTG ATC ACC GAC TTC GAC GCC AGC CA - #G GAC CGC ATC GAC CTC11388Thr Asp Leu Ile Thr Asp Phe Asp Ala Ser Gl - #n Asp Arg Ile Asp Leu# 470- TCC GCG CTG GGC TTC ACC GGT CTG GGC AAC GG - #T TAT GAC GGC ACC CTG11436Ser Ala Leu Gly Phe Thr Gly Leu Gly Asn Gl - #y Tyr Asp Gly Thr Leu# 485- GCG GTG ACC ACC GGT TCC GGC GGC ACC CGC AC - #C TAC CTG AAG AGC TAC11484Ala Val Thr Thr Gly Ser Gly Gly Thr Arg Th - #r Tyr Leu Lys Ser Tyr# 500- GAG GTG GAC GCC CAG GGC CGG CGT TTC GAA AT - #C GCC CTG GAC GGC AAC11532Glu Val Asp Ala Gln Gly Arg Arg Phe Glu Il - #e Ala Leu Asp Gly Asn505 5 - #10 5 - #15 5 -#20- TTC GTC GGC CAG TTC AAC GAT GGC AAC CTG TT - #G TTC GAC GCC GCT CCG11580Phe Val Gly Gln Phe Asn Asp Gly Asn Leu Le - #u Phe Asp Ala Ala Pro# 535- GTC ACC GGT ACC GAG GGC AAC GAC AAC CTG TC - #C GGC ACC GAT GCC GGG11628Val Thr Gly Thr Glu Gly Asn Asp Asn Leu Se - #r Gly Thr Asp Ala Gly# 550- GAA ACC CTC CTG GGC TAC GGC GGC AAC GAC AC - #C CTC AAC GGC GGG GCC11676Glu Thr Leu Leu Gly Tyr Gly Gly Asn Asp Th - #r Leu Asn Gly Gly Ala# 565- GGC AAC GAC ATC CTG GTC GGC GGC GCC GGG CG - #C GAC ACC CTG ACC GGC11724Gly Asn Asp Ile Leu Val Gly Gly Ala Gly Ar - #g Asp Thr Leu Thr Gly# 580- GGC GCC GGG GCG GAC GTG TTC CGC TTC GAG GC - #G CTG TCC GAC AGC CAG11772Gly Ala Gly Ala Asp Val Phe Arg Phe Glu Al - #a Leu Ser Asp Ser Gln585 5 - #90 5 - #95 6 -#00- CGC AAC TAC ACC GCC GGC GAC AAC CAG GGC GA - #T TAC ATC ATC GAC TTC11820Arg Asn Tyr Thr Ala Gly Asp Asn Gln Gly As - #p Tyr Ile Ile Asp Phe# 615- GCC GTG GGC GAA GAC AGG ATC GAC GTA TCG GC - #G CTG GGT TAC ACC GGG11868Ala Val Gly Glu Asp Arg Ile Asp Val Ser Al - #a Leu Gly Tyr Thr Gly# 630- CTG GGC AAC GGC CGC AAC GGC ACC CTC GCC GT - #G GTG CTC AAC AGC GCC11916Leu Gly Asn Gly Arg Asn Gly Thr Leu Ala Va - #l Val Leu Asn Ser Ala# 645- GGC GAC CGC ACC TAC GTG AAG AGC TAC GAC AC - #T GAC GCC AAC GGC TAT11964Gly Asp Arg Thr Tyr Val Lys Ser Tyr Asp Th - #r Asp Ala Asn Gly Tyr# 660- AAC TTC GAG CTT TCC CTC GCG GGC AAC TAC CA - #G GGG CTG CTC GGC GCC12012Asn Phe Glu Leu Ser Leu Ala Gly Asn Tyr Gl - #n Gly Leu Leu Gly Ala665 6 - #70 6 - #75 6 -#80- GAA CAG TTC GTC TTC GCC ACG CCC CCG GAA CA - #G GCG ACC ATC GAG GGA12060Glu Gln Phe Val Phe Ala Thr Pro Pro Glu Gl - #n Ala Thr Ile Glu Gly# 695- ACC GAC GGC AAC GAC AGC TTG CAA GGG ACC GG - #G GCC GAC GAA CTG CTC12108Thr Asp Gly Asn Asp Ser Leu Gln Gly Thr Gl - #y Ala Asp Glu Leu Leu# 710- CTC GGT CTG GGC GGC CGG GAC AGC CTG AAC GG - #C GGC GCC GGC GAC GAT12156Leu Gly Leu Gly Gly Arg Asp Ser Leu Asn Gl - #y Gly Ala Gly Asp Asp# 725- GTC CTG GAT GGC GGG GCG GAG CGC GAC ACC CT - #G ACC GGC GGC ACG GGG12204Val Leu Asp Gly Gly Ala Glu Arg Asp Thr Le - #u Thr Gly Gly Thr Gly# 740- GCC GAC ACC TTC CTG TTC TCC GCG CGT ACC GA - #C AGC TAC CGC ACC GAC12252Ala Asp Thr Phe Leu Phe Ser Ala Arg Thr As - #p Ser Tyr Arg Thr Asp745 7 - #50 7 - #55 7 -#60- AGC GCC AGC TTC ACC GAC CTG ATC ACC GAC TT - #C GAT CCC GCC CAG GAT12300Ser Ala Ser Phe Thr Asp Leu Ile Thr Asp Ph - #e Asp Pro Ala Gln Asp# 775- CGC ATC GAC CTG TCC GGC CTG GGC TTC AGC GG - #T TTC GGC AAC GGC TAC12348Arg Ile Asp Leu Ser Gly Leu Gly Phe Ser Gl - #y Phe Gly Asn Gly Tyr# 790- GAC GGC ACC CTG CTG CTG CAG GTC AAC GCC GC - #G GGC ACC CGC ACC TAC12396Asp Gly Thr Leu Leu Leu Gln Val Asn Ala Al - #a Gly Thr Arg Thr Tyr# 805- CTG AAG AGC CTC GAG GCC GAT GCC GAC GGC CA - #G CGC TTC GAG ATC GCC12444Leu Lys Ser Leu Glu Ala Asp Ala Asp Gly Gl - #n Arg Phe Glu Ile Ala# 820- CTG GAC GGC GAC TTC AGC GGC CAG TTG GAC AG - #C GGC AAC GTG ATC TTC12492Leu Asp Gly Asp Phe Ser Gly Gln Leu Asp Se - #r Gly Asn Val Ile Phe825 8 - #30 8 - #35 8 -#40- GAG GCC GGC GTG TTC AAT GCC AAG GAC TTC GG - #C GCG CTG GGC GAC GGC12540Glu Ala Gly Val Phe Asn Ala Lys Asp Phe Gl - #y Ala Leu Gly Asp Gly# 855- GCC AGC GAC GAC CGG CCG GCC ATC CAG GCG GC - #G ATC GAC GCC GCC TAC12588Ala Ser Asp Asp Arg Pro Ala Ile Gln Ala Al - #a Ile Asp Ala Ala Tyr# 870- (2) INFORMATION FOR SEQ ID NO:2:- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 553 ami - #no acids (B) TYPE: amino acid (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:- Met Asp Tyr Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Val Ser# 15- Asp Asp Arg Ala Ser Ile Gln Ala Ala Ile As - #p Ala Ala Tyr Ala Ala# 30- Gly Gly Gly Thr Val Tyr Leu Pro Ala Gly Gl - #u Tyr Arg Val Ser Ala# 45- Ala Gly Glu Pro Gly Asp Gly Cys Leu Met Le - #u Lys Asp Gly Val Tyr# 60- Leu Ala Gly Ala Gly Met Gly Glu Thr Val Il - #e Lys Leu Ile Asp Gly# 80- Ser Asp Gln Lys Ile Thr Gly Met Val Arg Se - #r Ala Tyr Gly Glu Glu# 95- Thr Ser Asn Phe Gly Met Arg Asp Leu Thr Le - #u Asp Gly Asn Arg Asp# 110- Asn Thr Ser Gly Lys Val Asp Gly Trp Phe As - #n Gly Tyr Ile Pro Gly# 125- Gly Asp Gly Ala Asp Arg Asp Val Thr Ile Gl - #u Arg Val Glu Val Arg# 140- Glu Met Ser Gly Tyr Gly Phe Asp Pro His Gl - #u Gln Thr Ile Asn Leu145 1 - #50 1 - #55 1 -#60- Thr Ile Arg Asp Ser Val Ala His Asp Asn Gl - #y Leu Asp Gly Phe Val# 175- Ala Asp Tyr Leu Val Asp Ser Val Phe Glu As - #n Asn Val Ala Tyr Ala# 190- Asn Asp Arg His Gly Phe Asn Val Val Thr Se - #r Thr His Asp Phe Val# 205- Met Thr Asn Asn Val Ala Tyr Gly Asn Gly Se - #r Ser Gly Leu Val Val# 220- Gln Arg Gly Leu Glu Asp Leu Ala Leu Pro Se - #r Asn Ile Leu Ile Asp225 2 - #30 2 - #35 2 -#40- Gly Gly Ala Tyr Tyr Asp Asn Ala Arg Glu Gl - #y Val Leu Leu Lys Met# 255- Thr Ser Asp Ile Thr Leu Gln Asn Ala Asp Il - #e His Gly Asn Gly Ser# 270- Ser Gly Val Arg Val Tyr Gly Ala Gln Asp Va - #l Gln Ile Leu Asp Asn# 285- Gln Ile His Asp Asn Ala Gln Ala Ala Ala Va - #l Pro Glu Val Leu Leu# 300- Gln Ser Phe Asp Asp Thr Ala Gly Ala Ser Gl - #y Thr Tyr Tyr Thr Thr305 3 - #10 3 - #15 3 -#20- Leu Asn Thr Arg Ile Glu Gly Asn Thr Ile Se - #r Gly Ser Ala Asn Ser# 335- Thr Tyr Gly Ile Gln Glu Arg Asn Asp Gly Th - #r Asp Tyr Ser Ser Leu# 350- Ile Asp Asn Asp Ile Ala Gly Val Gln Gln Pr - #o Ile Gln Leu Tyr Gly# 365- Pro His Ser Thr Val Ser Gly Glu Pro Gly Al - #a Thr Pro Gln Gln Pro# 380- Ser Thr Gly Ser Asp Gly Glu Pro Leu Val Gl - #y Gly Asp Thr Asp Asp385 3 - #90 3 - #95 4 -#00- Gln Leu Gln Gly Gly Ser Gly Ala Asp Arg Le - #u Asp Gly Gly Ala Gly# 415- Asp Asp Ile Leu Asp Gly Gly Ala Gly Arg As - #p Arg Leu Ser Gly Gly# 430- Ala Gly Ala Asp Thr Phe Val Phe Ser Ala Ar - #g Glu Asp Ser Tyr Arg# 445- Thr Asp Thr Ala Val Phe Asn Asp Leu Ile Le - #u Asp Phe Glu Ala Ser# 460- Glu Asp Arg Ile Asp Leu Ser Ala Leu Gly Ph - #e Ser Gly Leu Gly Asp465 4 - #70 4 - #75 4 -#80- Gly Tyr Gly Gly Thr Leu Leu Leu Lys Thr As - #n Ala Glu Gly Thr Arg# 495- Thr Tyr Leu Lys Ser Phe Glu Ala Asp Ala Gl - #u Gly Arg Arg Phe Glu# 510- Val Ala Leu Asp Gly Asp His Thr Gly Asp Le - #u Ser Ala Ala Asn Val# 525- Val Phe Ala Ala Thr Gly Thr Thr Thr Glu Le - #u Glu Val Leu Gly Asp# 540- Ser Gly Thr Gln Ala Gly Ala Ile Val545 5 - #50- (2) INFORMATION FOR SEQ ID NO:3:- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1403 am - #ino acids (B) TYPE: amino acid (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:- Met Asp Tyr Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Val Ser# 15- Asp Asp Thr Ala Ala Ile Gln Ala Ala Ile As - #p Ala Ala His Ala Ala# 30- Gly Gly Gly Thr Val Tyr Leu Pro Ala Gly Gl - #u Tyr Arg Val Ser Gly# 45- Gly Glu Glu Pro Ser Asp Gly Cys Leu Thr Il - #e Lys Ser Asn Val His# 60- Ile Val Gly Ala Gly Met Gly Glu Thr Val Il - #e Lys Met Val Asp Gly# 80- Trp Thr Gln Asn Val Thr Gly Met Val Arg Se - #r Ala Tyr Gly Glu Glu# 95- Thr Ser Asn Phe Gly Met Ser Asp Leu Thr Le - #u Asp Gly Asn Arg Asp# 110- Asn Leu Ser Ala Lys Val Asp Gly Trp Phe As - #n Gly Tyr Ile Pro Gly# 125- Gln Asp Gly Ala Asp Arg Asp Val Thr Leu Gl - #u Arg Val Glu Ile Arg# 140- Glu Met Ser Gly Tyr Gly Phe Asp Pro His Gl - #u Gln Thr Ile Asn Leu145 1 - #50 1 - #55 1 -#60- Thr Ile Arg Asp Ser Val Ala His Asp Asn Se - #r Leu Asp Gly Phe Val# 175- Ala Asp Tyr Gln Val Gly Gly Val Phe Glu As - #n Asn Val Ser Tyr Asn# 190- Asn Asp Arg His Gly Phe Asn Ile Val Thr Se - #r Thr Asn Asp Phe Val# 205- Leu Ser Asn Asn Val Ala Tyr Gly Asn Gly Gl - #y Ala Gly Leu Val Val# 220- Gln Arg Gly Ser Tyr Asp Leu Pro His Pro Ty - #r Asp Ile Leu Ile Asp225 2 - #30 2 - #35 2 -#40- Gly Gly Ala Tyr Tyr Asp Asn Ala Leu Glu Gl - #y Val Gln Leu Lys Met# 255- Ala His Asp Val Thr Leu Gln Asn Ala Glu Il - #e Tyr Gly Asn Gly Leu# 270- Tyr Gly Val Arg Val Tyr Gly Ala Gln Asp Va - #l Gln Ile Leu Asp Asn# 285- Gln Ile His Asp Asn Ser Gln Asn Gly Ala Ty - #r Ala Glu Val Leu Leu# 300- Gln Ser Tyr Asp Asp Thr Ala Gly Val Ser Gl - #y Asn Phe Tyr Val Thr305 3 - #10 3 - #15 3 -#20- Thr Gly Thr Trp Leu Glu Gly Asn Val Ile Se - #r Gly Ser Ala Asn Ser# 335- Thr Tyr Gly Ile Gln Glu Arg Ala Asp Gly Th - #r Asp Tyr Ser Ser Leu# 350- Tyr Ala Asn Ser Ile Asp Gly Val Gln Thr Gl - #y Ala Val Arg Leu Tyr# 365- Gly Ala Asn Ser Thr Val Ser Ser Gln Ser Gl - #y Ser Gly Gln Gln Ala# 380- Thr Leu Glu Gly Ser Ala Gly Asn Asp Ala Le - #u Ser Gly Thr Glu Ala385 3 - #90 3 - #95 4 -#00- His Glu Thr Leu Leu Gly Gln Ala Gly Asp As - #p Arg Leu Asn Gly Asp# 415- Ala Gly Asn Asp Ile Leu Asp Gly Gly Ala Gl - #y Arg Asp Asn Leu Thr# 430- Gly Gly Ala Gly Ala Asp Thr Phe Arg Phe Se - #r Ala Arg Thr Asp Ser# 445- Tyr Arg Thr Asp Ser Ala Ser Phe Asn Asp Le - #u Ile Thr Asp Phe Asp# 460- Ala Asp Glu Asp Ser Ile Asp Leu Ser Ala Le - #u Gly Phe Thr Gly Leu465 4 - #70 4 - #75 4 -#80- Gly Asp Gly Tyr Asn Gly Thr Leu Leu Leu Ly - #s Thr Asn Ala Glu Gly# 495- Thr Arg Thr Tyr Leu Lys Ser Tyr Glu Ala As - #p Ala Gln Gly Arg Arg# 510- Phe Glu Ile Ala Leu Asp Gly Asn Phe Thr Gl - #y Leu Phe Asn Asp Asn# 525- Asn Leu Leu Phe Asp Ala Ala Pro Ala Thr Gl - #y Thr Glu Gly Ser Asp# 540- Asn Leu Leu Gly Thr Asp Ala Gly Glu Thr Le - #u Leu Gly Tyr Gly Gly545 5 - #50 5 - #55 5 -#60- Asn Asp Thr Leu Asn Gly Gly Ala Gly Asp As - #p Ile Leu Val Gly Gly# 575- Ala Gly Arg Asp Ser Leu Thr Gly Gly Ala Gl - #y Ala Asp Val Phe Arg# 590- Phe Asp Ala Leu Ser Asp Ser Gln Arg Asn Ty - #r Thr Thr Gly Asp Asn# 605- Gln Ala Asp Arg Ile Leu Asp Phe Asp Pro Th - #r Leu Asp Arg Ile Asp# 620- Val Ser Ala Leu Gly Phe Thr Gly Leu Gly As - #n Gly Arg Asn Gly Thr625 6 - #30 6 - #35 6 -#40- Leu Ala Val Val Leu Asn Ser Ala Gly Asp Ar - #g Thr Asp Leu Lys Ser# 655- Tyr Asp Thr Asp Ala Asn Gly Tyr Ser Phe Gl - #u Leu Ser Leu Ala Gly# 670- Asn Tyr Gln Gly Gln Leu Ser Ala Glu Gln Ph - #e Val Phe Ala Thr Ser# 685- Gln Gly Gly Gln Met Thr Ile Ile Glu Gly Th - #r Asp Gly Asn Asp Thr# 700- Leu Gln Gly Thr Glu Ala Asn Glu Arg Leu Le - #u Gly Leu Asp Gly Arg705 7 - #10 7 - #15 7 -#20- Asp Asn Leu Asn Gly Gly Ala Gly Asp Asp Il - #e Leu Asp Gly Gly Ala# 735- Gly Arg Asp Thr Leu Thr Gly Gly Thr Gly Al - #a Asp Thr Phe Leu Phe# 750- Ser Thr Arg Thr Asp Ser Tyr Arg Thr Asp Se - #r Ala Ser Phe Asn Asp# 765- Leu Ile Thr Asp Phe Asp Pro Thr Gln Asp Ar - #g Ile Asp Leu Ser Gly# 780- Leu Gly Phe Ser Gly Phe Gly Asn Gly Tyr As - #p Gly Thr Leu Leu Leu785 7 - #90 7 - #95 8 -#00- Gln Val Asn Ala Ala Gly Thr Arg Thr Tyr Le - #u Lys Ser Phe Glu Ala# 815- Asp Ala Asn Gly Gln Arg Phe Glu Ile Ala Le - #u Asp Gly Asp Phe Ser# 830- Gly Gln Leu Asp Ser Gly Asn Val Ile Phe Gl - #u Pro Ala Val Phe Asn# 845- Ala Lys Asp Phe Gly Ala Leu Gly Asp Gly Al - #a Ser Asp Asp Arg Pro# 860- Ala Ile Gln Ala Ala Ile Asp Ala Ala Tyr Al - #a Ala Gly Gly Gly Thr865 8 - #70 8 - #75 8 -#80- Val Tyr Leu Pro Ala Gly Glu Tyr Arg Val Se - #r Pro Thr Gly Glu Pro# 895- Gly Asp Gly Cys Leu Met Leu Lys Asp Gly Va - #l Tyr Leu Ala Gly Asp# 910- Gly Ile Gly Glu Thr Val Ile Lys Leu Ile As - #p Gly Ser Asp Gln Lys# 925- Ile Thr Gly Met Val Arg Ser Ala Tyr Gly Gl - #u Glu Thr Ser Asn Phe# 940- Gly Met Ser Asp Leu Thr Leu Asp Gly Asn Ar - #g Asp Asn Thr Ser Gly945 9 - #50 9 - #55 9 -#60- Lys Val Asp Gly Trp Phe Asn Gly Tyr Ile Pr - #o Gly Gln Asp Gly Ala# 975- Asp Arg Asn Val Thr Ile Glu Arg Val Glu Il - #e Arg Glu Met Ser Gly# 990- Tyr Gly Phe Asp Pro His Glu Gln Thr Ile As - #n Leu Thr Ile Arg Asp# 10050- Ser Val Ala His Asp Asn Gly Leu Asp Gly Ph - #e Val Ala Asp Tyr Leu# 10205- Val Asp Ser Val Phe Glu Asn Asn Val Ala Ty - #r Asn Asn Asp Arg His# 10401030 - # 1035- Gly Phe Asn Ile Val Thr Ser Thr Tyr Asp Ph - #e Val Met Thr Asn Asn# 10550- Val Ala Tyr Gly Asn Gly Gly Ala Gly Leu Th - #r Ile Gln Arg Gly Ser# 10705- Glu Asp Leu Ala Gln Pro Thr Asp Ile Leu Il - #e Asp Gly Gly Ala Tyr# 10850- Tyr Asp Asn Ala Leu Glu Gly Val Leu Phe Ly - #s Met Thr Asn Asn Val# 11005- Thr Leu Gln Asn Ala Glu Ile Tyr Gly Asn Gl - #y Ser Ser Gly Val Arg# 11201110 - # 1115- Leu Tyr Gly Thr Glu Asp Val Gln Ile Leu As - #p Asn Gln Ile His Asp# 11350- Asn Ser Gln Asn Gly Thr Tyr Pro Glu Val Le - #u Leu Gln Ala Phe Asp# 11505- Asp Ser Gln Val Thr Gly Glu Leu Tyr Glu Th - #r Leu Asn Thr Arg Ile# 11650- Glu Gly Asn Leu Ile Asp Ala Ser Asp Asn Al - #a Asn Tyr Ala Val Arg# 11805- Glu Arg Asp Asp Gly Ser Asp Tyr Thr Thr Le - #u Val Asp Asn Asp Ile# 12001190 - # 1195- Ser Gly Gly Gln Val Ala Ser Val Gln Leu Se - #r Gly Ala His Ser Ser# 12150- Leu Ser Gly Gly Thr Val Glu Val Pro Gln Gl - #y Thr Asp Gly Asn Asp# 12305- Val Leu Val Gly Ser Asp Ala Asn Asp Gln Le - #u Tyr Gly Gly Ala Gly# 12450- Asp Asp Arg Leu Asp Gly Gly Ala Gly Asp As - #p Leu Leu Asp Gly Gly# 12605- Ala Gly Arg Asp Asp Leu Thr Gly Gly Thr Gl - #y Ala Asp Thr Phe Val# 12801270 - # 1275- Phe Ala Ala Arg Thr Asp Ser Tyr Arg Thr As - #p Ala Gly Val Phe Asn# 12950- Asp Leu Ile Leu Asp Phe Asp Ala Ser Glu As - #p Arg Ile Asp Leu Ser# 13105- Ala Leu Gly Phe Ser Gly Phe Gly Asp Gly Ty - #r Asn Gly Thr Leu Leu# 13250- Val Gln Leu Ser Ser Ala Gly Thr Arg Thr Ty - #r Leu Lys Ser Tyr Glu# 13405- Glu Asp Leu Glu Gly Arg Arg Phe Glu Val Al - #a Leu Asp Gly Asp His# 13601350 - # 1355- Thr Gly Asp Leu Ser Ala Ala Asn Val Val Ph - #e Ala Asp Asp Gly Ser# 13750- Ala Ala Val Ala Ser Ser Asp Pro Ala Ala Th - #r Gln Leu Glu Val Val# 13905- Gly Ser Ser Gly Thr Gln Thr Asp Gln Leu Al - #a# 1400- (2) INFORMATION FOR SEQ ID NO:4:- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 997 ami - #no acids (B) TYPE: amino acid (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:- Met Asp Tyr Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Val Ser# 15- Asp Asp Thr Ala Ala Ile Gln Ala Ala Ile As - #p Ala Ala Tyr Ala Ala# 30- Gly Gly Gly Thr Val Tyr Leu Pro Ala Gly Gl - #u Tyr Arg Val Ser Gly# 45- Gly Glu Glu Pro Ser Asp Gly Cys Leu Thr Il - #e Lys Ser Asn Val His# 60- Ile Val Gly Ala Gly Met Gly Glu Thr Val Il - #e Lys Leu Val Asp Gly# 80- Trp Asp Gln Asp Val Thr Gly Ile Val Arg Se - #r Ala Tyr Gly Glu Glu# 95- Thr Ser Asn Phe Gly Met Ser Asp Leu Thr Le - #u Asp Gly Asn Arg Asp# 110- Asn Thr Ser Gly Lys Val Asp Gly Trp Phe As - #n Gly Tyr Ile Pro Gly# 125- Glu Asp Gly Ala Asp Arg Asp Val Thr Leu Gl - #u Arg Val Glu Ile Arg# 140- Glu Met Ser Gly Tyr Gly Phe Asp Pro His Gl - #u Gln Thr Ile Asn Leu145 1 - #50 1 - #55 1 -#60- Thr Ile Arg Asp Ser Val Ala His Asp Asn Gl - #y Leu Asp Gly Phe Val# 175- Ala Asp Phe Gln Ile Gly Gly Val Phe Glu As - #n Asn Val Ser Tyr Asn# 190- Asn Asp Arg His Gly Phe Asn Ile Val Thr Se - #r Thr Asn Asp Phe Val# 205- Leu Ser Asn Asn Val Ala Tyr Gly Asn Gly Gl - #y Ala Gly Leu Val Val# 220- Gln Arg Gly Ser Ser Asp Val Ala His Pro Ty - #r Asp Ile Leu Ile Asp225 2 - #30 2 - #35 2 -#40- Gly Gly Ala Tyr Tyr Asp Asn Gly Leu Glu Gl - #y Val Gln Ile Lys Met# 255- Ala His Asp Val Thr Leu Gln Asn Ala Glu Il - #e Tyr Gly Asn Gly Leu# 270- Tyr Gly Val Arg Val Tyr Gly Ala Glu Asp Va - #l Gln Ile Leu Asp Asn# 285- Tyr Ile His Asp Asn Ser Gln Asn Gly Ser Ty - #r Ala Glu Ile Leu Leu# 300- Gln Ser Tyr Asp Asp Thr Ala Gly Val Ser Gl - #y Asn Phe Tyr Thr Thr305 3 - #10 3 - #15 3 -#20- Thr Gly Thr Trp Ile Glu Gly Asn Thr Ile Va - #l Gly Ser Ala Asn Ser# 335- Thr Tyr Gly Ile Gln Glu Arg Asp Asp Gly Th - #r Asp Tyr Ser Ser Leu# 350- Tyr Ala Asn Ser Val Ser Asn Val Gln Asn Gl - #y Ser Val Arg Leu Tyr# 365- Gly Ala Asn Ser Val Val Ser Asp Leu Pro Gl - #y Thr Gly Gln Gln Ala# 380- Thr Leu Glu Gly Thr Ala Gly Asn Asp Thr Le - #u Gly Gly Ser Asp Ala385 3 - #90 3 - #95 4 -#00- His Glu Thr Leu Leu Gly Leu Asp Gly Asn As - #p Arg Leu Asn Gly Gly# 415- Ala Gly Asn Asp Ile Leu Asp Gly Gly Ala Gl - #y Arg Asp Asn Leu Thr# 430- Gly Gly Ala Gly Ala Asp Leu Phe Arg Val Se - #r Ala Arg Thr Asp Ser# 445- Tyr Arg Thr Asp Ser Ala Ser Phe Asn Asp Le - #u Ile Thr Asp Phe Asp# 460- Ala Ser Gln Asp Arg Ile Asp Leu Ser Ala Le - #u Gly Phe Thr Gly Leu465 4 - #70 4 - #75 4 -#80- Gly Asp Gly Tyr Asn Gly Thr Leu Leu Leu Gl - #n Val Ser Ala Asp Gly# 495- Ser Arg Thr Tyr Leu Lys Ser Leu Glu Ala As - #p Ala Glu Gly Arg Arg# 510- Phe Glu Ile Ala Leu Asp Gly Asn Phe Ala Gl - #y Leu Leu Gly Ala Gly# 525- Asn Leu Leu Phe Glu Arg Thr Ala Ile Glu Gl - #y Asp Ala Gly Asp Asn# 540- Ala Leu Leu Gly Thr Ser Ala Ala Glu Thr Le - #u Leu Gly His Ala Gly545 5 - #50 5 - #55 5 -#60- Asn Asp Thr Leu Asp Gly Gly Ala Gly Asp As - #p Ile Leu Val Gly Gly# 575- Ala Gly Arg Asp Ser Leu Thr Gly Gly Ala Gl - #y Ala Asp Val Phe Arg# 590- Phe Asp Ala Leu Ser Asp Ser Gln Arg Asn Ty - #r Asp Ile Gly Asp Asn# 605- Gln Gly Asp Arg Ile Ala Asp Phe Ala Val Gl - #y Glu Asp Lys Leu Asp# 620- Val Ser Ala Leu Gly Phe Thr Gly Leu Gly As - #p Gly Tyr Asn Gly Thr625 6 - #30 6 - #35 6 -#40- Leu Ala Leu Val Leu Asn Ser Ala Gly Asp Ar - #g Thr Tyr Val Lys Ser# 655- Tyr Glu Asn Gly Ala Asp Gly Tyr Arg Phe Gl - #u Phe Ser Leu Asp Gly# 670- Asn Tyr Leu Glu Leu Leu Gly Asn Glu Asp Ph - #e Ile Phe Ala Thr Pro# 685- Ser Gly Gln Gln Leu Leu Glu Gly Ser Ala Gl - #y Asn Asp Ser Leu Gln# 700- Gly Thr Ala Ala Asp Glu Val Ile His Gly Gl - #y Gly Gly Arg Asp Thr705 7 - #10 7 - #15 7 -#20- Leu Ala Gly Gly Ala Gly Ala Asp Val Phe Ar - #g Phe Ser Glu Leu Thr# 735- Asp Ser Tyr Arg Asp Ser Ala Ser Tyr Ala As - #p Leu Ile Thr Asp Phe# 750- Asp Ala Ser Glu Asp Arg Ile Asp Leu Ser Gl - #y Leu Gly Phe Ser Gly# 765- Leu Gly Asn Gly Tyr Gly Gly Thr Leu Ala Le - #u Gln Val Asn Ser Ala# 780- Gly Thr Arg Thr Tyr Leu Lys Ser Phe Glu Th - #r Asn Ala Ala Gly Glu785 7 - #90 7 - #95 8 -#00- Arg Phe Glu Ile Ala Leu Asp Gly Asp Leu Se - #r Ala Leu Gly Gly Ala# 815- Asn Leu Ile Leu Asp Ala Arg Thr Val Leu Al - #a Gly Gly Asp Gly Asn# 830- Asp Thr Leu Ser Gly Ser Ser Ala Ala Glu Gl - #u Leu Leu Gly Gly Val# 845- Gly Asn Asp Ser Leu Asp Gly Gly Ala Gly As - #n Asp Ile Leu Asp Gly# 860- Gly Ala Gly Arg Asp Thr Leu Ser Gly Gly Se - #r Gly Ser Asp Ile Phe865 8 - #70 8 - #75 8 -#80- Arg Phe Gly Gly Ala Leu Asp Ser Phe Arg As - #n Tyr Ala Ser Gly Thr# 895- Asn Gly Thr Asp Ser Ile Thr Asp Phe Thr Pr - #o Gly Glu Asp Leu Ile# 910- Asp Leu Ser Val Leu Gly Tyr Thr Gly Leu Gl - #y Asp Gly Tyr Asn Gly# 925- Thr Leu Ala Ile Val Leu Asn Asp Ala Gly Th - #r Lys Thr Tyr Leu Lys# 940- Asn Arg Glu Ser Asp Ala Glu Gly Asn Gln Ph - #e Glu Ile Ala Leu Glu945 9 - #50 9 - #55 9 -#60- Gly Asn His Ala Asp Gln Leu Asp Ala Ser As - #p Phe Ile Phe Ala Thr# 975- Ala Ala Ala Thr Thr Gly Ile Glu Val Val Gl - #y Gly Ser Gly Thr Gln# 990- Thr Asp Gln Leu Ala 995- (2) INFORMATION FOR SEQ ID NO:5:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 872 amino (B) TYPE: amino acid (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:- Met Asp Phe Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Ala Ser# 15- Asp Asp Thr Ala Ala Ile Gln Ala Ala Ile As - #p Ala Ala His Ala Ala# 30- Gly Gly Gly Thr Val Tyr Leu Pro Ala Gly Gl - #u Tyr Arg Val Ser Gly# 45- Gly Glu Glu Pro Ser Asp Gly Ala Leu Thr Il - #e Lys Ser Asn Val Tyr# 60- Ile Val Gly Ala Gly Met Gly Glu Thr Val Il - #e Lys Met Val Asp Gly# 80- Trp Thr Gln Asn Val Thr Gly Met Val Arg Se - #r Ala Tyr Gly Glu Glu# 95- Thr Ser Asn Phe Gly Met Ser Asp Leu Thr Le - #u Asp Gly Asn Arg Asp# 110- Asn Leu Ser Ala Lys Val Asp Gly Trp Phe As - #n Gly Tyr Ile Pro Gly# 125- Gln Asp Gly Ala Asp Arg Asp Val Thr Leu Gl - #u Arg Val Glu Ile Arg# 140- Glu Met Ser Gly Tyr Gly Phe Asp Pro His Gl - #u Gln Thr Ile Asn Leu145 1 - #50 1 - #55 1 -#60- Thr Ile Arg Asp Ser Val Ala His Asp Asn Gl - #y Leu Asp Gly Phe Val# 175- Ala Asp Tyr Gln Val Gly Gly Val Phe Glu As - #n Asn Val Ser Tyr Asn# 190- Asn Asp Arg His Gly Phe Asn Ile Val Thr Se - #r Thr Asn Asp Phe Val# 205- Leu Ser Asn Asn Val Ala Tyr Gly Asn Gly Gl - #y Ala Gly Leu Val Val# 220- Gln Arg Gly Ser Tyr Asp Leu Pro His Pro Ty - #r Asp Ile Leu Ile Asp225 2 - #30 2 - #35 2 -#40- Gly Gly Ala Tyr Tyr Asp Asn Ala Leu Glu Gl - #y Val Gln Leu Lys Met# 255- Thr His Asp Val Thr Leu Gln Asn Ala Glu Il - #e Tyr Gly Asn Gly Leu# 270- Tyr Gly Val Arg Val Tyr Gly Ala Gln Asp Va - #l Gln Leu Leu Asp Asn# 285- Gln Ile His Asp Asn Ser Gln Asn Gly Ala Ty - #r Ala Glu Val Leu Leu# 300- Gln Ser Tyr Asp Asp Thr Ala Gly Val Ser Gl - #y Asn Phe Tyr Val Thr305 3 - #10 3 - #15 3 -#20- Thr Gly Thr Trp Leu Glu Gly Asn Val Ile Se - #r Gly Ser Ala Asn Ser# 335- Thr Phe Gly Ile Gln Glu Arg Ala Asp Gly Th - #r Asp Tyr Ser Ser Leu# 350- Tyr Ala Asn Thr Ile Asp Gly Val Gln Asn Gl - #y Thr Val Arg Leu Tyr# 365- Gly Ala Asn Ser Thr Val Ser Glu Gln Pro Se - #r Ser Gly Gln Gln Ala# 380- Thr Leu Glu Gly Thr Ala Gly Asn Asp Val Le - #u Ser Gly Thr Gly Ala385 3 - #90 3 - #95 4 -#00- His Glu Leu Ile Leu Gly Leu Ala Gly Asn As - #p Arg Leu Asp Gly Gly# 415- Ala Gly Asp Asp Thr Leu Asp Gly Gly Ala Gl - #y Arg Asp Thr Leu Thr# 430- Gly Gly Ala Gly Ala Asp Thr Phe Arg Phe Se - #r Ala Arg Glu Asp Ser# 445- His Arg Thr Asp Ser Ala Ser Phe Thr Asp Le - #u Ile Thr Asp Phe Asp# 460- Ala Ser Gln Asp Arg Ile Asp Leu Ser Ala Le - #u Gly Phe Thr Gly Leu465 4 - #70 4 - #75 4 -#80- Gly Asn Gly Tyr Asp Gly Thr Leu Ala Val Th - #r Thr Gly Ser Gly Gly# 495- Thr Arg Thr Tyr Leu Lys Ser Tyr Glu Val As - #p Ala Gln Gly Arg Arg# 510- Phe Glu Ile Ala Leu Asp Gly Asn Phe Val Gl - #y Gln Phe Asn Asp Gly# 525- Asn Leu Leu Phe Asp Ala Ala Pro Val Thr Gl - #y Thr Glu Gly Asn Asp# 540- Asn Leu Ser Gly Thr Asp Ala Gly Glu Thr Le - #u Leu Gly Tyr Gly Gly545 5 - #50 5 - #55 5 -#60- Asn Asp Thr Leu Asn Gly Gly Ala Gly Asn As - #p Ile Leu Val Gly Gly# 575- Ala Gly Arg Asp Thr Leu Thr Gly Gly Ala Gl - #y Ala Asp Val Phe Arg# 590- Phe Glu Ala Leu Ser Asp Ser Gln Arg Asn Ty - #r Thr Ala Gly Asp Asn# 605- Gln Gly Asp Tyr Ile Ile Asp Phe Ala Val Gl - #y Glu Asp Arg Ile Asp# 620- Val Ser Ala Leu Gly Tyr Thr Gly Leu Gly As - #n Gly Arg Asn Gly Thr625 6 - #30 6 - #35 6 -#40- Leu Ala Val Val Leu Asn Ser Ala Gly Asp Ar - #g Thr Tyr Val Lys Ser# 655- Tyr Asp Thr Asp Ala Asn Gly Tyr Asn Phe Gl - #u Leu Ser Leu Ala Gly# 670- Asn Tyr Gln Gly Leu Leu Gly Ala Glu Gln Ph - #e Val Phe Ala Thr Pro# 685- Pro Glu Gln Ala Thr Ile Glu Gly Thr Asp Gl - #y Asn Asp Ser Leu Gln# 700- Gly Thr Gly Ala Asp Glu Leu Leu Leu Gly Le - #u Gly Gly Arg Asp Ser705 7 - #10 7 - #15 7 -#20- Leu Asn Gly Gly Ala Gly Asp Asp Val Leu As - #p Gly Gly Ala Glu Arg# 735- Asp Thr Leu Thr Gly Gly Thr Gly Ala Asp Th - #r Phe Leu Phe Ser Ala# 750- Arg Thr Asp Ser Tyr Arg Thr Asp Ser Ala Se - #r Phe Thr Asp Leu Ile# 765- Thr Asp Phe Asp Pro Ala Gln Asp Arg Ile As - #p Leu Ser Gly Leu Gly# 780- Phe Ser Gly Phe Gly Asn Gly Tyr Asp Gly Th - #r Leu Leu Leu Gln Val785 7 - #90 7 - #95 8 -#00- Asn Ala Ala Gly Thr Arg Thr Tyr Leu Lys Se - #r Leu Glu Ala Asp Ala# 815- Asp Gly Gln Arg Phe Glu Ile Ala Leu Asp Gl - #y Asp Phe Ser Gly Gln# 830- Leu Asp Ser Gly Asn Val Ile Phe Glu Ala Gl - #y Val Phe Asn Ala Lys# 845- Asp Phe Gly Ala Leu Gly Asp Gly Ala Ser As - #p Asp Arg Pro Ala Ile# 860- Gln Ala Ala Ile Asp Ala Ala Tyr865 8 - #70- (2) INFORMATION FOR SEQ ID NO:6:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 15 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: peptide- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:- Met Asp Tyr Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Val# 15- (2) INFORMATION FOR SEQ ID NO:7:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1155 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:- ATGGATTACA ACGTCAAGGA TTTCGGGGCG CTGGGCGATG GCGTCAGCGA CG - #ACACGGCG 60- GCCATCCAGG CGGCGATCGA TGCCGCCTAC GCGGCCGGCG GCGGCACCGT CT - #ACCTGCCG 120- GCCGGCGAGT ACCGGGTCAG CGGCGGCGAG GAGCCTTCCG ACGGCTGCCT GA - #CCATCAAG 180- AGCAACGTCT ATATCGTCGG CGCCGGGATG GGCGAGACGG TGATCAAGCT GG - #TCGACGGC 240- TGGGAGCAGA ACGTCACCGG CATGGTGCGC TCGGCCTACG GCGAGGAGAC CA - #GCAACTTC 300- GGCATGAGCG ACCTGACCCT CGACGGCAAC CGCGACAACA CCAGCGGCAA GG - #TCGACGGC 360- TGGTTCAACG GCTACATCCC CGGCCAGGAC GGCGCCGACC GCGACGTGAC CC - #TGGAGCGG 420- GTGGAAATCC GCGAGATGTC CGGCTACGGT TTCGATCCGC ACGAGCAGAC CA - #TCAACCTG 480- ACGATCCGCG ACAGCGTGGC CCACGACAAC GGCCTCGACG GCTTCGTCGC CG - #ACTACCAG 540- GTCGGCGGGG TGTTCGAGAA CAACGTCTCG TACAACAACG ACCGCCACGG CT - #TCAACATC 600- GTCACCAGCA CCAACGACTT CGTCCTGAGC AACAACGTCG CCTACGGCAA CG - #GCGGCGCC 660- GGCCTGGTGG TGCAGCGCGG CTCGTACGAC CTGGCCCAGC CCTACGACAT CC - #TGATCGAC 720- GGCGGCGCCT ACTACGACAA CGCCCTGGAA GGCGTGCAGC TCAAGATGAC CC - #ACGACGTC 780- ACCCTGCAGA ACGCCGAGAT CTACGGCAAC GGCCTCTACG GGGTGCGCGT CT - #ACGGCGCC 840- CAGGACGTGC AGATCCTCGA CAACCAGATC CACGACAATT CGCAGAACGG CG - #CCTATGCC 900- GAAGTCCTGC TGCAGTCCTA CGACGACACC GCCGGGGTGT CCGGCAACTT CT - #ACGCCACC 960- ACCGGCACCT GGATCGAAGG CAACATCATC AGCGGCTCGG CCAACTCCAC CT - #ACGGCATC1020- CAGGAGCGCG ACGACGGCAC CGACTACAGC AGCCTCTACG CCAACAACAT CG - #GCGGTGTG1080- CAGAACGGGT CGGTACGGCT GTACGGCGCC AACTCGACGG TTTCCGGCCA GC - #CCGGCACC1140# 1155- (2) INFORMATION FOR SEQ ID NO:8:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 385 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: peptide- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:- Met Asp Phe Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Ala Ser# 15- Asp Asp Thr Ala Ala Ile Gln Ala Ala Ile As - #p Ala Ala Tyr Ala Ala# 30- Gly Gly Gly Thr Val Tyr Leu Pro Ala Gly Gl - #u Tyr Arg Val Ser Gly# 45- Gly Glu Glu Pro Ser Asp Gly Cys Leu Thr Il - #e Lys Ser Asn Val Tyr# 60- Ile Val Gly Ala Gly Met Gly Glu Thr Val Il - #e Lys Leu Val Asp Gly#80- Trp Asp Gln Asn Val Thr Gly Met Val Arg Se - #r Ala Tyr Gly Glu Glu# 95- Thr Ser Asn Phe Gly Met Ser Asp Leu Thr Le - #u Asp Gly Asn Arg Asp# 110- Asn Thr Ser Gly Lys Val Asp Gly Trp Phe As - #n Gly Tyr Ile Pro Gly# 125- Gln Asp Gly Ala Asp Arg Asp Val Thr Leu Gl - #u Arg Val Glu Ile Arg# 140- Glu Met Ser Gly Tyr Gly Phe Asp Pro His Gl - #u Gln Thr Ile Asn Leu145 1 - #50 1 - #55 1 -#60- Thr Ile Arg Asp Ser Val Ala His Asp Asn Gl - #y Leu Asp Gly Phe Val# 175- Ala Asp Tyr Gln Val Gly Gly Val Phe Glu As - #n Asn Val Ser Tyr Asn# 190- Asn Asp Arg His Gly Phe Asn Ile Val Thr Se - #r Thr Asn Asp Phe Val# 205- Leu Ser Asn Asn Val Ala Tyr Gly Asn Gly Gl - #y Ala Gly Leu Val Val# 220- Gln Arg Gly Ser Glu Asp Leu Ala His Pro Ty - #r Asp Ile Leu Ile Asp225 2 - #30 2 - #35 2 -#40- Gly Gly Ala Tyr Tyr Asp Asn Ala Leu Glu Gl - #y Val Gln Leu Lys Met# 255- Thr His Asp Val Thr Leu Gln Asn Ala Glu Il - #e Tyr Gly Asn Gly Leu# 270- Tyr Gly Val Arg Val Tyr Gly Ala Gln Asp Va - #l Gln Ile Leu Asp Asn# 285- Gln Ile His Asp Asn Ser Gln Asn Gly Ala Ty - #r Ala Glu Val Leu Leu# 300- Gln Ser Tyr Asp Asp Thr Ala Gly Val Ser Gl - #y Asn Phe Tyr Thr Thr305 3 - #10 3 - #15 3 -#20- Thr Gly Thr Trp Ile Glu Gly Asn Thr Ile Se - #r Gly Ser Ala Asn Ser# 335- Thr Tyr Gly Ile Gln Glu Arg Ala Asp Gly Th - #r Asp Tyr Ser Ser Leu# 350- Tyr Ala Asn Asp Ile Asp Gly Val Gln Thr Gl - #y Ser Val Arg Leu Tyr# 365- Gly Ala Asn Ser Thr Val Ser Gly Gln Pro Gl - #y Ser Gly Gln Gln Ala# 380- Thr- 385- (2) INFORMATION FOR SEQ ID NO:9:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 459 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:- CTCGAAGGCA CCGACGGCAA CGACACGCTG CGCGGCACCG AGGCCGACGA GA - #CGCTCCTC 60- GGCCAGGCCG GCAACGACCG CCTGAACGGC GGCGCCGGCG ACGACATCCT CG - #ACGGCGGC 120- GCCGGGCGCG ACACCCTGAC CGGCGGCGCG GGCGCCGACA CCTTCCGCTT CT - #CCGCGCGG 180- ACCGACAGCT ACCGCACCGA CAGCGCCGGC GACAGCTTCA ACGACCTGAT CA - #CCGACTTC 240- GACGCCAGCG AGGACCGCAT CGACCTGTCC GCGCTGGGCT TCACCGGGCT GG - #GCGACGGC 300- TACAACGGCA CCCTGCTGCT GGTGCTCAAC GCCGCCGGCA CCCGCACCTA CC - #TGAAGAGC 360- TACGAGGCGG ACGCCGAGGG CCAGCGCTTC GAGATCGCCC TGGACGGCAA CT - #ACACCGGC 420# 459 ACTT GGTCTTCGCC GCGGCCGCG- (2) INFORMATION FOR SEQ ID NO:10:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 153 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: peptide- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:- Leu Glu Gly Thr Asp Gly Asn Asp Thr Leu Se - #r Gly Thr Asp Ala His# 15- Glu Thr Leu Leu Gly Leu Ala Gly Asn Asp Ar - #g Leu Asn Gly Gly Ala# 30- Gly Asp Asp Ile Leu Asp Gly Gly Ala Gly Ar - #g Asp Thr Leu Thr Gly# 45- Gly Ala Gly Ala Asp Thr Phe Arg Phe Ser Al - #a Arg Thr Asp Ser Tyr# 60- Arg Thr Asp Ser Ala Gly Asp Ser Phe Asn As - #p Leu Ile Thr Asp Phe#80- Asp Ala Ser Glu Asp Arg Ile Asp Leu Ser Al - #a Leu Gly Phe Thr Gly# 95- Leu Gly Asp Gly Tyr Asn Gly Thr Leu Leu Le - #u Gln Leu Asn Ser Ala# 110- Gly Thr Arg Thr Tyr Leu Lys Ser Tyr Glu Al - #a Asp Ala Glu Gly Arg# 125- Arg Phe Glu Ile Ala Leu Asp Gly Asn Phe Th - #r Gly Leu Leu Gly Ala# 140- Glu Asn Phe Ile Phe Ala Thr Thr Pro145 1 - #50- (2) INFORMATION FOR SEQ ID NO:11:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 9 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: peptide- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:- Leu Xaa Gly Gly Ala Gly Xaa Asp Xaa1 5- (2) INFORMATION FOR SEQ ID NO:12:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 6 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: peptide- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:- Ala Arg Arg Arg Arg Ser1 5- (2) INFORMATION FOR SEQ ID NO:13:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 7 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: peptide- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:- Ala Arg Arg Arg Ala Arg Arg1 5- (2) INFORMATION FOR SEQ ID NO:14:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 4 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: peptide- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:- Arg Ala Arg Ser- (2) INFORMATION FOR SEQ ID NO:15:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 4 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: peptide- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:- Ala Arg Arg Arg1- (2) INFORMATION FOR SEQ ID NO:16:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 20 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: other nucleic acid#= "Oligonucleotide"PTION: /desc- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:# 20 ARGA- (2) INFORMATION FOR SEQ ID NO:17:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1176 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:- ATGGATTACA ACGTCAAGGA TTTCGGTGCA TTGGGCGACG GCGTCAGCGA CG - #ACCGGGCC 60- TCCATCCAGG CGGCGATCGA TGCCGCCTAC GCCGCCGGTG GCGGTACCGT CT - #ACCTGCCG 120- GCCGGCGAGT ACCGGGTCAG CGCCGCCGGG GAGCCGGGCG ACGGCTGCCT GA - #TGCTCAAG 180- GACGGCGTCT ACCTGGCCGG TGCCGGCATG GGCGAGACGG TGATCAAGCT GA - #TCGACGGC 240- TCCGACCAGA AGATCACCGG CATGGTCCGC TCGGCCTACG GCGAGGAAAC CA - #GCAACTTC 300- GGCATGCGCG ACCTGACCCT CGACGGCAAC CGCGACAACA CCAGCGGCAA GG - #TCGACGGC 360- TGGTTCAACG GCTATATCCC CGGCGGGGAC GGCGCCGACC GCGACGTGAC CA - #TCGAGCGG 420- GTGGAGGTCC GCGAGATGTC CGGCTACGGC TTCGACCCCC ACGAGCAGAC CA - #TCAACCTG 480- ACGATCCGCG ACAGCGTGGC CCACGACAAC GGCCTCGACG GCTTCGTCGC CG - #ACTACCTG 540- GTCGACAGCG TGTTCGAGAA CAACGTCGCC TACGCCAACG ACCGCCACGG CT - #TCAACGTG 600- GTCACCAGCA CCCACGATTT CGTCATGACC AACAACGTCG CCTACGGCAA CG - #GCAGCAGC 660- GGCCTGGTGG TGCAGCGGGG TCTGGAGGAC CTCGCGCTGC CCAGCAACAT CC - #TGATCGAC 720- GGCGGCGCCT ACTACGACAA CGCCCGCGAA GGCGTGCTGC TCAAGATGAC CA - #GCGACATC 780- ACCCTGCAGA ACGCCGATAT CCACGGCAAC GGCTCCTCCG GGGTGCGCGT CT - #ACGGCGCC 840- CAGGACGTGC AGATCCTCGA TAACCAGATC CACGACAACG CGCAGGCGGC CG - #CCGTGCCC 900- GAGGTCCTGC TGCAGTCCTT CGACGATACC GCCGGGGCGT CCGGCACCTA CT - #ACACGACC 960- CTGAACACCC GGATCGAGGG CAACACCATC AGCGGCTCGG CCAACTCCAC CT - #ACGGCATC1020- CAGGAGCGCA ACGACGGCAC CGACTACAGC AGCCTGATCG ACAACGACAT CG - #CCGGGGTG1080- CAACAGCCCA TCCAACTGTA CGGACCTCAC TCGACGGTAT CCGGCGAACC CG - #GCGCGACA1140# 1176 CGGG AAGCGACGGC GAGCCA- (2) INFORMATION FOR SEQ ID NO:18:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1155 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:- ATGGATTACA ACGTCAAGGA TTTCGGAGCA CTGGGCGATG GCGTCAGCGA CG - #ACACGGCG 60- GCCATCCAGG CGGCGATCGA CGCCGCCCAC GCGGCGGGCG GCGGCACCGT CT - #ACCTGCCG 120- GCCGGCGAAT ATCGGGTCAG CGGCGGCGAG GAGCCTTCCG ATGGTTGTCT GA - #CCATCAAG 180- AGCAACGTCC ATATCGTCGG CGCCGGGATG GGCGAGACGG TGATCAAGAT GG - #TCGACGGC 240- TGGACGCAGA ACGTCACCGG CATGGTGCGC TCGGCCTACG GCGAGGAAAC CA - #GCAACTTC 300- GGCATGAGCG ACCTGACCCT CGACGGCAAC CGCGACAACC TGTCCGCCAA GG - #TCGACGGC 360- TGGTTCAACG GCTACATCCC CGGCCAGGAC GGCGCCGATC GCGACGTGAC CC - #TGGAGCGG 420- GTGGAAATCC GCGAGATGTC CGGCTACGGT TTCGACCCCC ACGAGCAGAC CA - #TCAACCTG 480- ACGATCCGCG ACAGCGTGGC CCACGACAAC AGCCTCGACG GCTTCGTCGC CG - #ACTACCAG 540- GTCGGCGGGG TGTTCGAGAA CAACGTCTCG TACAACAACG ACCGCCACGG CT - #TCAACATC 600- GTCACCAGCA CCAACGACTT CGTCCTGAGC AACAACGTCG CCTACGGCAA CG - #GCGGCGCC 660- GGCCTGGTGG TGCAGCGCGG CTCGTACGAC CTGCCCCATC CCTACGACAT CC - #TGATCGAC 720- GGCGGCGCCT ACTACGACAA CGCCTTGGAA GGCGTGCAGC TCAAGATGGC CC - #ACGACGTC 780- ACCCTGCAGA ACGCCGAGAT CTACGGCAAC GGCCTGTACG GGGTGCGCGT CT - #ACGGCGCC 840- CAGGACGTGC AGATCCTCGA CAACCAGATC CACGACAATT CGCAGAACGG CG - #CCTATGCC 900- GAAGTCCTGC TGCAGTCCTA CGACGACACC GCCGGGGTGT CCGGCAACTT TT - #ACGTCACC 960- ACCGGCACCT GGCTCGAAGG CAACGTCATC AGCGGCTCGG CCAATTCCAC CT - #ACGGCATC1020- CAGGAGCGCG CCGACGGCAC CGACTACAGC AGCCTCTACG CCAACAGCAT CG - #ACGGTGTG1080- CAGACCGGGG CGGTACGGCT GTATGGCGCC AACTCGACGG TTTCCAGCCA GT - #CCGGCAGT1140# 1155- (2) INFORMATION FOR SEQ ID NO:19:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1143 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:- GTGTTCAATG CCAAGGACTT CGGCGCGCTG GGCGACGGCG CCAGCGACGA CC - #GGCCGGCC 60- ATCCAGGCGG CGATCGACGC CGCCTACGCG GCCGGTGGCG GCACCGTCTA CC - #TGCCGGCC 120- GGCGAGTACC GGGTCAGCCC CACCGGGGAG CCGGGCGACG GCTGCCTGAT GC - #TCAAGGAC 180- GGCGTCTACC TGGCCGGCGA CGGCATAGGC GAAACGGTCA TCAAGCTGAT CG - #ACGGCTCC 240- GACCAGAAGA TCACCGGCAT GGTGCGCTCG GCCTATGGCG AAGAGACCAG CA - #ACTTCGGC 300- ATGAGCGACC TGACCCTCGA CGGCAACCGC GACAACACCA GCGGCAAGGT CG - #ACGGCTGG 360- TTCAACGGCT ACATCCCCGG CCAGGACGGC GCCGACCGCA ACGTGACCAT CG - #AGCGGGTG 420- GAAATCCGCG AGATGTCCGG CTATGGCTTC GATCCGCACG AGCAGACCAT CA - #ACCTGACG 480- ATCCGCGACA GCGTGGCCCA CGACAACGGC CTCGACGGCT TCGTCGCCGA CT - #ACCTGGTC 540- GACAGCGTGT TCGAGAACAA CGTCGCCTAC AACAACGACC GCCACGGCTT CA - #ACATCGTC 600- ACCAGCACCT ACGATTTCGT CATGACCAAC AACGTCGCCT ACGGCAACGG CG - #GCGCCGGC 660- CTGACGATCC AGCGGGGCTC GGAGGACCTG GCCCAGCCGA CCGATATCCT GA - #TCGACGGC 720- GGCGCCTACT ACGACAACGC CCTGGAAGGC GTGCTGTTCA AGATGACCAA CA - #ACGTCACC 780- CTGCAGAACG CCGAGATCTA CGGCAACGGC TCCTCCGGCG TGCGCCTGTA CG - #GCACGGAG 840- GACGTGCAGA TCCTCGACAA CCAGATCCAC GACAATTCGC AGAACGGCAC CT - #ATCCGGAA 900- GTCCTGCTGC AGGCCTTCGA CGACAGCCAG GTCACCGGTG AGCTGTACGA GA - #CCCTGAAC 960- ACCCGGATCG AAGGCAATCT CATCGACGCT TCGGACAACG CCAACTATGC GG - #TGCGCGAG1020- CGCGACGACG GCAGCGACTA CACCACGCTC GTGGACAACG ACATCAGCGG CG - #GCCAGGTC1080- GCCTCGGTGC AGCTTTCCGG CGCCCATTCG AGTCTTTCCG GCGGCACCGT CG - #AAGTGCCG1140# 1143- (2) INFORMATION FOR SEQ ID NO:20:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1155 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:- ATGGATTACA ACGTCAAAGA TTTCGGGGCG CTGGGCGATG GCGTCAGCGA CG - #ATACGGCC 60- GCCATCCAGG CGGCGATCGA TGCCGCCTAC GCGGCCGGCG GCGGCACCGT CT - #ACCTGCCG 120- GCCGGCGAAT ACCGGGTCAG CGGCGGCGAG GAGCCTTCCG ATGGTTGCCT GA - #CCATCAAG 180- AGCAACGTCC ATATCGTCGG CGCGGGGATG GGCGAGACGG TCATCAAGCT GG - #TCGACGGC 240- TGGGATCAGG ACGTCACCGG CATCGTCCGC TCGGCCTACG GCGAGGAGAC CA - #GCAACTTC 300- GGCATGAGCG ACCTGACCCT CGACGGCAAC CGCGACAACA CCAGCGGCAA GG - #TCGACGGC 360- TGGTTCAACG GCTACATTCC CGGCGAGGAC GGCGCCGACC GCGACGTGAC CC - #TGGAGCGG 420- GTGGAAATCC GTGAAATGTC CGGTTACGGT TTCGATCCGC ACGAGCAGAC CA - #TCAACCTG 480- ACGATCCGCG ACAGCGTGGC CCACGACAAC GGCCTCGACG GCTTCGTCGC CG - #ATTTCCAG 540- ATCGGCGGGG TGTTCGAGAA CAACGTCTCG TACAACAACG ACCGCCACGG CT - #TCAACATC 600- GTCACCAGCA CCAACGACTT CGTCCTGAGC AACAACGTCG CCTACGGCAA CG - #GCGGCGCC 660- GGCCTGGTGG TGCAGCGCGG CTCGTCCGAC GTGGCGCACC CCTACGACAT CC - #TGATCGAC 720- GGCGGCGCCT ACTACGACAA CGGCCTGGAA GGCGTGCAGA TCAAGATGGC CC - #ACGACGTC 780- ACCCTGCAGA ACGCCGAGAT CTACGGCAAC GGCCTATACG GGGTGCGCGT CT - #ACGGCGCC 840- GAGGATGTGC AGATCCTCGA CAACTACATC CACGACAATT CGCAGAACGG TT - #CCTACGCG 900- GAAATCCTCC TGCAGTCCTA CGACGATACC GCCGGGGTGT CCGGCAATTT CT - #ACACCACC 960- ACCGGCACCT GGATCGAAGG CAACACCATC GTCGGCTCGG CCAACTCCAC CT - #ATGGCATC1020- CAGGAGCGCG ACGACGGCAC CGACTACAGC AGCCTCTACG CCAACAGCGT CA - #GCAATGTG1080- CAGAACGGCT CGGTGCGCCT CTACGGCGCC AACTCCGTCG TCTCCGACCT GC - #CCGGCACC1140# 1155- (2) INFORMATION FOR SEQ ID NO:21:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 1155 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:- ATGGACTTCA ACGTCAAAGA TTTCGGGGCA CTGGGCGATG GCGCCAGCGA CG - #ACACGGCG 60- GCCATCCAGG CGGCGATCGA TGCCGCCCAC GCGGCGGGCG GCGGCACCGT CT - #ACCTGCCG 120- GCTGGCGAGT ATCGGGTCAG CGGCGGCGAG GAGCCTTCCG ACGGCGCGCT GA - #CCATCAAG 180- AGCAACGTCT ATATCGTCGG CGCCGGGATG GGCGAGACGG TGATCAAGAT GG - #TCGACGGC 240- TGGACGCAGA ACGTCACCGG CATGGTGCGC TCGGCCTATG GCGAGGAGAC CA - #GCAACTTC 300- GGCATGAGCG ACCTGACCCT CGACGGCAAC CGCGACAACC TGTCCGCCAA GG - #TCGACGGC 360- TGGTTCAACG GCTACATTCC CGGCCAGGAC GGTGCCGATC GCGACGTGAC CC - #TGGAGCGG 420- GTGGAAATCC GCGAAATGTC CGGTTACGGT TTCGATCCGC ACGAGCAGAC CA - #TCAACCTG 480- ACGATCCGCG ACAGCGTGGC CCACGACAAC GGCCTCGACG GCTTCGTCGC CG - #ACTACCAG 540- GTCGGCGGGG TGTTCGAGAA CAACGTCTCG TACAACAACG ACCGCCACGG CT - #TCAACATC 600- GTCACCAGCA CCAACGACTT CGTCCTGAGC AACAACGTCG CCTACGGCAA CG - #GCGGCGCC 660- GGCCTGGTGG TGCAGCGCGG CTCGTACGAC CTGCCCCATC CCTACGACAT CC - #TGATCGAC 720- GGCGGCGCCT ACTACGACAA CGCCTTGGAA GGCGTGCAGC TCAAGATGAC CC - #ACGACGTC 780- ACCCTGCAGA ACGCCGAGAT CTATGGCAAC GGCCTGTACG GGGTGCGCGT CT - #ACGGCGCC 840- CAGGACGTGC AACTCCTCGA TAACCAGATC CACGACAATT CGCAGAACGG CG - #CCTATGCC 900- GAAGTCCTGC TGCAGTCCTA CGACGACACC GCCGGGGTGT CCGGCAACTT TT - #ACGTCACC 960- ACCGGCACCT GGCTCGAAGG CAACGTCATC AGCGGCTCGG CCAATTCCAC CT - #TCGGCATC1020- CAGGAGCGCG CCGACGGCAC CGACTACAGC AGCCTTTACG CCAATACCAT CG - #ACGGCGTG1080- CAGAACGGGA CGGTACGGCT GTATGGCGCC AACTCCACGG TTTCCGAGCA GC - #CCAGCAGC1140# 1155- (2) INFORMATION FOR SEQ ID NO:22:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 87 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:- GTGTTCAATG CCAAGGACTT CGGCGCGCTG GGCGACGGCG CCAGCGACGA CC - #GGCCGGCC 60# 87 ACGC CGCCTAC- (2) INFORMATION FOR SEQ ID NO:23:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 392 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:- Met Asp Tyr Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Val Ser# 15- Asp Asp Arg Ala Ser Ile Gln Ala Ala Ile As - #p Ala Ala Tyr Ala Ala# 30- Gly Gly Gly Thr Val Tyr Leu Pro Ala Gly Gl - #u Tyr Arg Val Ser Ala# 45- Ala Gly Glu Pro Gly Asp Gly Cys Leu Met Le - #u Lys Asp Gly Val Tyr# 60- Leu Ala Gly Ala Gly Met Gly Glu Thr Val Il - #e Lys Leu Ile Asp Gly#80- Ser Asp Gln Lys Ile Thr Gly Met Val Arg Se - #r Ala Tyr Gly Glu Glu# 95- Thr Ser Asn Phe Gly Met Arg Asp Leu Thr Le - #u Asp Gly Asn Arg Asp# 110- Asn Thr Ser Gly Lys Val Asp Gly Trp Phe As - #n Gly Tyr Ile Pro Gly# 125- Gly Asp Gly Ala Asp Arg Asp Val Thr Ile Gl - #u Arg Val Glu Val Arg# 140- Glu Met Ser Gly Tyr Gly Phe Asp Pro His Gl - #u Gln Thr Ile Asn Leu145 1 - #50 1 - #55 1 -#60- Thr Ile Arg Asp Ser Val Ala His Asp Asn Gl - #y Leu Asp Gly Phe Val# 175- Ala Asp Tyr Leu Val Asp Ser Val Phe Glu As - #n Asn Val Ala Tyr Ala# 190- Asn Asp Arg His Gly Phe Asn Val Val Thr Se - #r Thr His Asp Phe Val# 205- Met Thr Asn Asn Val Ala Tyr Gly Asn Gly Se - #r Ser Gly Leu Val Val# 220- Gln Arg Gly Leu Glu Asp Leu Ala Leu Pro Se - #r Asn Ile Leu Ile Asp225 2 - #30 2 - #35 2 -#40- Gly Gly Ala Tyr Tyr Asp Asn Ala Arg Glu Gl - #y Val Leu Leu Lys Met# 255- Thr Ser Asp Ile Thr Leu Gln Asn Ala Asp Il - #e His Gly Asn Gly Ser# 270- Ser Gly Val Arg Val Tyr Gly Ala Gln Asp Va - #l Gln Ile Leu Asp Asn# 285- Gln Ile His Asp Asn Ala Gln Ala Ala Ala Va - #l Pro Glu Val Leu Leu# 300- Gln Ser Phe Asp Asp Thr Ala Gly Ala Ser Gl - #y Thr Tyr Tyr Thr Thr305 3 - #10 3 - #15 3 -#20- Leu Asn Thr Arg Ile Glu Gly Asn Thr Ile Se - #r Gly Ser Ala Asn Ser# 335- Thr Tyr Gly Ile Gln Glu Arg Asn Asp Gly Th - #r Asp Tyr Ser Ser Leu# 350- Ile Asp Asn Asp Ile Ala Gly Val Gln Gln Pr - #o Ile Gln Leu Tyr Gly# 365- Pro His Ser Thr Val Ser Gly Glu Pro Gly Al - #a Thr Pro Gln Gln Pro# 380- Ser Thr Gly Ser Asp Gly Glu Pro385 3 - #90- (2) INFORMATION FOR SEQ ID NO:24:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 385 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:- Met Asp Tyr Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Val Ser# 15- Asp Asp Thr Ala Ala Ile Gln Ala Ala Ile As - #p Ala Ala His Ala Ala# 30- Gly Gly Gly Thr Val Tyr Leu Pro Ala Gly Gl - #u Tyr Arg Val Ser Gly# 45- Gly Glu Glu Pro Ser Asp Gly Cys Leu Thr Il - #e Lys Ser Asn Val His# 60- Ile Val Gly Ala Gly Met Gly Glu Thr Val Il - #e Lys Met Val Asp Gly#80- Trp Thr Gln Asn Val Thr Gly Met Val Arg Se - #r Ala Tyr Gly Glu Glu# 95- Thr Ser Asn Phe Gly Met Ser Asp Leu Thr Le - #u Asp Gly Asn Arg Asp# 110- Asn Leu Ser Ala Lys Val Asp Gly Trp Phe As - #n Gly Tyr Ile Pro Gly# 125- Gln Asp Gly Ala Asp Arg Asp Val Thr Leu Gl - #u Arg Val Glu Ile Arg# 140- Glu Met Ser Gly Tyr Gly Phe Asp Pro His Gl - #u Gln Thr Ile Asn Leu145 1 - #50 1 - #55 1 -#60- Thr Ile Arg Asp Ser Val Ala His Asp Asn Se - #r Leu Asp Gly Phe Val# 175- Ala Asp Tyr Gln Val Gly Gly Val Phe Glu As - #n Asn Val Ser Tyr Asn# 190- Asn Asp Arg His Gly Phe Asn Ile Val Thr Se - #r Thr Asn Asp Phe Val# 205- Leu Ser Asn Asn Val Ala Tyr Gly Asn Gly Gl - #y Ala Gly Leu Val Val# 220- Gln Arg Gly Ser Tyr Asp Leu Pro His Pro Ty - #r Asp Ile Leu Ile Asp225 2 - #30 2 - #35 2 -#40- Gly Gly Ala Tyr Tyr Asp Asn Ala Leu Glu Gl - #y Val Gln Leu Lys Met# 255- Ala His Asp Val Thr Leu Gln Asn Ala Glu Il - #e Tyr Gly Asn Gly Leu# 270- Tyr Gly Val Arg Val Tyr Gly Ala Gln Asp Va - #l Gln Ile Leu Asp Asn# 285- Gln Ile His Asp Asn Ser Gln Asn Gly Ala Ty - #r Ala Glu Val Leu Leu# 300- Gln Ser Tyr Asp Asp Thr Ala Gly Val Ser Gl - #y Asn Phe Tyr Val Thr305 3 - #10 3 - #15 3 -#20- Thr Gly Thr Trp Leu Glu Gly Asn Val Ile Se - #r Gly Ser Ala Asn Ser# 335- Thr Tyr Gly Ile Gln Glu Arg Ala Asp Gly Th - #r Asp Tyr Ser Ser Leu# 350- Tyr Ala Asn Ser Ile Asp Gly Val Gln Thr Gl - #y Ala Val Arg Leu Tyr# 365- Gly Ala Asn Ser Thr Val Ser Ser Gln Ser Gl - #y Ser Gly Gln Gln Ala# 380- Thr- 385- (2) INFORMATION FOR SEQ ID NO:25:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 381 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:- Val Phe Asn Ala Lys Asp Phe Gly Ala Leu Gl - #y Asp Gly Ala Ser Asp# 15- Asp Arg Pro Ala Ile Gln Ala Ala Ile Asp Al - #a Ala Tyr Ala Ala Gly# 30- Gly Gly Thr Val Tyr Leu Pro Ala Gly Glu Ty - #r Arg Val Ser Pro Thr# 45- Gly Glu Pro Gly Asp Gly Cys Leu Met Leu Ly - #s Asp Gly Val Tyr Leu# 60- Ala Gly Asp Gly Ile Gly Glu Thr Val Ile Ly - #s Leu Ile Asp Gly Ser#80- Asp Gln Lys Ile Thr Gly Met Val Arg Ser Al - #a Tyr Gly Glu Glu Thr# 95- Ser Asn Phe Gly Met Ser Asp Leu Thr Leu As - #p Gly Asn Arg Asp Asn# 110- Thr Ser Gly Lys Val Asp Gly Trp Phe Asn Gl - #y Tyr Ile Pro Gly Gln# 125- Asp Gly Ala Asp Arg Asn Val Thr Ile Glu Ar - #g Val Glu Ile Arg Glu# 140- Met Ser Gly Tyr Gly Phe Asp Pro His Glu Gl - #n Thr Ile Asn Leu Thr145 1 - #50 1 - #55 1 -#60- Ile Arg Asp Ser Val Ala His Asp Asn Gly Le - #u Asp Gly Phe Val Ala# 175- Asp Tyr Leu Val Asp Ser Val Phe Glu Asn As - #n Val Ala Tyr Asn Asn# 190- Asp Arg His Gly Phe Asn Ile Val Thr Ser Th - #r Tyr Asp Phe Val Met# 205- Thr Asn Asn Val Ala Tyr Gly Asn Gly Gly Al - #a Gly Leu Thr Ile Gln# 220- Arg Gly Ser Glu Asp Leu Ala Gln Pro Thr As - #p Ile Leu Ile Asp Gly225 2 - #30 2 - #35 2 -#40- Gly Ala Tyr Tyr Asp Asn Ala Leu Glu Gly Va - #l Leu Phe Lys Met Thr# 255- Asn Asn Val Thr Leu Gln Asn Ala Glu Ile Ty - #r Gly Asn Gly Ser Ser# 270- Gly Val Arg Leu Tyr Gly Thr Glu Asp Val Gl - #n Ile Leu Asp Asn Gln# 285- Ile His Asp Asn Ser Gln Asn Gly Thr Tyr Pr - #o Glu Val Leu Leu Gln# 300- Ala Phe Asp Asp Ser Gln Val Thr Gly Glu Le - #u Tyr Glu Thr Leu Asn305 3 - #10 3 - #15 3 -#20- Thr Arg Ile Glu Gly Asn Leu Ile Asp Ala Se - #r Asp Asn Ala Asn Tyr# 335- Ala Val Arg Glu Arg Asp Asp Gly Ser Asp Ty - #r Thr Thr Leu Val Asp# 350- Asn Asp Ile Ser Gly Gly Gln Val Ala Ser Va - #l Gln Leu Ser Gly Ala# 365- His Ser Ser Leu Ser Gly Gly Thr Val Glu Va - #l Pro Gln# 380- (2) INFORMATION FOR SEQ ID NO:26:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 385 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:- Met Asp Tyr Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Val Ser# 15- Asp Asp Thr Ala Ala Ile Gln Ala Ala Ile As - #p Ala Ala Tyr Ala Ala# 30- Gly Gly Gly Thr Val Tyr Leu Pro Ala Gly Gl - #u Tyr Arg Val Ser Gly# 45- Gly Glu Glu Pro Ser Asp Gly Cys Leu Thr Il - #e Lys Ser Asn Val His# 60- Ile Val Gly Ala Gly Met Gly Glu Thr Val Il - #e Lys Leu Val Asp Gly#80- Trp Asp Gln Asp Val Thr Gly Ile Val Arg Se - #r Ala Tyr Gly Glu Glu# 95- Thr Ser Asn Phe Gly Met Ser Asp Leu Thr Le - #u Asp Gly Asn Arg Asp# 110- Asn Thr Ser Gly Lys Val Asp Gly Trp Phe As - #n Gly Tyr Ile Pro Gly# 125- Glu Asp Gly Ala Asp Arg Asp Val Thr Leu Gl - #u Arg Val Glu Ile Arg# 140- Glu Met Ser Gly Tyr Gly Phe Asp Pro His Gl - #u Gln Thr Ile Asn Leu145 1 - #50 1 - #55 1 -#60- Thr Ile Arg Asp Ser Val Ala His Asp Asn Gl - #y Leu Asp Gly Phe Val# 175- Ala Asp Phe Gln Ile Gly Gly Val Phe Glu As - #n Asn Val Ser Tyr Asn# 190- Asn Asp Arg His Gly Phe Asn Ile Val Thr Se - #r Thr Asn Asp Phe Val# 205- Leu Ser Asn Asn Val Ala Tyr Gly Asn Gly Gl - #y Ala Gly Leu Val Val# 220- Gln Arg Gly Ser Ser Asp Val Ala His Pro Ty - #r Asp Ile Leu Ile Asp225 2 - #30 2 - #35 2 -#40- Gly Gly Ala Tyr Tyr Asp Asn Gly Leu Glu Gl - #y Val Gln Ile Lys Met# 255- Ala His Asp Val Thr Leu Gln Asn Ala Glu Il - #e Tyr Gly Asn Gly Leu# 270- Tyr Gly Val Arg Val Tyr Gly Ala Glu Asp Va - #l Gln Ile Leu Asp Asn# 285- Tyr Ile His Asp Asn Ser Gln Asn Gly Ser Ty - #r Ala Glu Ile Leu Leu# 300- Gln Ser Tyr Asp Asp Thr Ala Gly Val Ser Gl - #y Asn Phe Tyr Thr Thr305 3 - #10 3 - #15 3 -#20- Thr Gly Thr Trp Ile Glu Gly Asn Thr Ile Va - #l Gly Ser Ala Asn Ser# 335- Thr Tyr Gly Ile Gln Glu Arg Asp Asp Gly Th - #r Asp Tyr Ser Ser Leu# 350- Tyr Ala Asn Ser Val Ser Asn Val Gln Asn Gl - #y Ser Val Arg Leu Tyr# 365- Gly Ala Asn Ser Val Val Ser Asp Leu Pro Gl - #y Thr Gly Gln Gln Ala# 380- Thr- 385- (2) INFORMATION FOR SEQ ID NO:27:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 385 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:- Met Asp Phe Asn Val Lys Asp Phe Gly Ala Le - #u Gly Asp Gly Ala Ser# 15- Asp Asp Thr Ala Ala Ile Gln Ala Ala Ile As - #p Ala Ala His Ala Ala# 30- Gly Gly Gly Thr Val Tyr Leu Pro Ala Gly Gl - #u Tyr Arg Val Ser Gly# 45- Gly Glu Glu Pro Ser Asp Gly Ala Leu Thr Il - #e Lys Ser Asn Val Tyr# 60- Ile Val Gly Ala Gly Met Gly Glu Thr Val Il - #e Lys Met Val Asp Gly#80- Trp Thr Gln Asn Val Thr Gly Met Val Arg Se - #r Ala Tyr Gly Glu Glu# 95- Thr Ser Asn Phe Gly Met Ser Asp Leu Thr Le - #u Asp Gly Asn Arg Asp# 110- Asn Leu Ser Ala Lys Val Asp Gly Trp Phe As - #n Gly Tyr Ile Pro Gly# 125- Gln Asp Gly Ala Asp Arg Asp Val Thr Leu Gl - #u Arg Val Glu Ile Arg# 140- Glu Met Ser Gly Tyr Gly Phe Asp Pro His Gl - #u Gln Thr Ile Asn Leu145 1 - #50 1 - #55 1 -#60- Thr Ile Arg Asp Ser Val Ala His Asp Asn Gl - #y Leu Asp Gly Phe Val# 175- Ala Asp Tyr Gln Val Gly Gly Val Phe Glu As - #n Asn Val Ser Tyr Asn# 190- Asn Asp Arg His Gly Phe Asn Ile Val Thr Se - #r Thr Asn Asp Phe Val# 205- Leu Ser Asn Asn Val Ala Tyr Gly Asn Gly Gl - #y Ala Gly Leu Val Val# 220- Gln Arg Gly Ser Tyr Asp Leu Pro His Pro Ty - #r Asp Ile Leu Ile Asp225 2 - #30 2 - #35 2 -#40- Gly Gly Ala Tyr Tyr Asp Asn Ala Leu Glu Gl - #y Val Gln Leu Lys Met# 255- Thr His Asp Val Thr Leu Gln Asn Ala Glu Il - #e Tyr Gly Asn Gly Leu# 270- Tyr Gly Val Arg Val Tyr Gly Ala Gln Asp Va - #l Gln Leu Leu Asp Asn# 285- Gln Ile His Asp Asn Ser Gln Asn Gly Ala Ty - #r Ala Glu Val Leu Leu# 300- Gln Ser Tyr Asp Asp Thr Ala Gly Val Ser Gl - #y Asn Phe Tyr Val Thr305 3 - #10 3 - #15 3 -#20- Thr Gly Thr Trp Leu Glu Gly Asn Val Ile Se - #r Gly Ser Ala Asn Ser# 335- Thr Phe Gly Ile Gln Glu Arg Ala Asp Gly Th - #r Asp Tyr Ser Ser Leu# 350- Tyr Ala Asn Thr Ile Asp Gly Val Gln Asn Gl - #y Thr Val Arg Leu Tyr# 365- Gly Ala Asn Ser Thr Val Ser Glu Gln Pro Se - #r Ser Gly Gln Gln Ala# 380- Thr- 385- (2) INFORMATION FOR SEQ ID NO:28:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 29 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:- Val Phe Asn Ala Lys Asp Phe Gly Ala Leu Gl - #y Asp Gly Ala Ser Asp# 15- Asp Arg Pro Ala Ile Gln Ala Ala Ile Asp Al - #a Ala Tyr# 25- (2) INFORMATION FOR SEQ ID NO:29:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 426 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:- CTGGTCGGCG GCGACACGGA CGACCAGCTC CAGGGCGGCT CCGGCGCCGA TC - #GCCTGGAC 60- GGCGGGGCCG GCGACGACAT CCTCGACGGC GGCGCCGGGC GCGACCGGCT GA - #GCGGCGGC 120- GCGGGCGCCG ACACCTTCGT GTTCTCCGCC CGCGAGGACA GCTACCGTAC CG - #ACACGGCG 180- GTGTTCAACG ACCTGATCCT CGACTTCGAG GCCAGCGAGG ATCGCATCGA CC - #TGTCCGCG 240- CTGGGCTTTT CCGGCCTGGG CGACGGCTAT GGCGGCACCC TGCTCCTGAA GA - #CCAACGCC 300- GAGGGCACGC GCACCTACCT GAAAAGCTTC GAGGCGGATG CCGAGGGACG GC - #GCTTCGAG 360- GTCGCCCTGG ACGGCGACCA CACGGGCGAT CTTTCCGCCG CCAATGTGGT CT - #TCGCCGCG 420# 426- (2) INFORMATION FOR SEQ ID NO:30:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 453 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:- CTCGAAGGCA GCGCGGGCAA CGATGCGCTG AGCGGGACCG AGGCCCACGA GA - #CGCTGCTC 60- GGCCAGGCCG GCGACGACCG CCTGAACGGC GATGCCGGCA ACGACATCCT CG - #ACGGCGGG 120- GCAGGGCGCG ACAACCTGAC CGGCGGCGCG GGCGCCGACA CCTTCCGCTT CT - #CCGCGCGC 180- ACCGACAGCT ACCGCACCGA CAGCGCCAGC TTCAACGACC TGATCACCGA CT - #TCGACGCC 240- GACGAGGACA GCATCGACCT GTCCGCGCTG GGCTTCACCG GCCTGGGCGA CG - #GCTACAAT 300- GGCACCCTGC TGCTGAAGAC CAACGCCGAG GGTACGCGCA CCTACCTGAA GA - #GCTACGAA 360- GCGGACGCCC AGGGCCGGCG CTTCGAGATC GCCCTGGACG GCAACTTCAC CG - #GTCTGTTC 420# 453 TGTT CGACGCCGCT CCG- (2) INFORMATION FOR SEQ ID NO:31:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 459 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:- GCCACCGGTA CCGAGGGCAG CGACAACCTG CTCGGCACCG ACGCCGGGGA AA - #CCCTCCTG 60- GGCTACGGCG GCAACGACAC CCTCAACGGC GGGGCCGGCG ACGACATCCT GG - #TCGGCGGC 120- GCCGGGCGCG ACAGCCTGAC CGGCGGCGCC GGGGCGGACG TGTTCCGCTT CG - #ACGCGCTG 180- TCCGACAGCC AGCGCAACTA CACCACCGGC GACAACCAGG CCGACCGCAT TC - #TCGACTTC 240- GACCCGACCC TGGACAGGAT CGACGTGTCG GCGCTGGGCT TCACCGGGCT GG - #GCAACGGC 300- CGCAACGGCA CCCTCGCCGT GGTGCTCAAC AGCGCCGGCG ACCGCACCGA TC - #TGAAGAGC 360- TACGACACCG ACGCCAACGG CTACAGCTTC GAGCTTTCCC TCGCGGGCAA CT - #ACCAGGGG 420# 459 AGTT CGTTTTCGCG ACGTCTCAG- (2) INFORMATION FOR SEQ ID NO:32:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 450 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:- ATCGAAGGCA CCGACGGCAA CGATACCTTG CAGGGCACCG AGGCCAACGA GC - #GGCTCCTC 60- GGCCTGGACG GCCGGGACAA CCTGAACGGC GGCGCCGGCG ACGACATCCT CG - #ACGGCGGA 120- GCGGGGCGCG ACACCCTGAC CGGCGGCACG GGGGCCGACA CCTTCCTGTT CT - #CCACGCGT 180- ACCGACAGCT ACCGCACCGA CAGCGCCAGC TTCAACGACC TGATCACCGA CT - #TCGATCCC 240- ACCCAGGACC GCATCGACCT GTCCGGCCTG GGCTTCAGCG GTTTCGGCAA CG - #GCTACGAC 300- GGCACCCTGC TGCTGCAGGT CAACGCCGCG GGCACCCGCA CCTACCTGAA GA - #GTTTCGAG 360- GCCGATGCCA ACGGCCAGCG CTTCGAGATC GCCCTGGACG GCGACTTCAG CG - #GCCAATTG 420# 450 TCTT CGAGCCCGCC- (2) INFORMATION FOR SEQ ID NO:33:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 447 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:- GGGACCGACG GCAACGACGT GCTGGTCGGC AGCGATGCCA ACGACCAGCT CT - #ACGGCGGA 60- GCCGGCGACG ACCGCCTGGA CGGCGGCGCC GGTGACGACC TGCTCGACGG CG - #GAGCGGGG 120- CGCGACGACC TGACCGGCGG CACGGGTGCC GACACCTTCG TGTTCGCCGC GC - #GTACCGAT 180- AGCTACCGCA CCGACGCGGG GGTGTTCAAC GACCTGATCC TCGACTTCGA CG - #CCAGCGAG 240- GACCGCATCG ACCTGTCCGC CCTGGGTTTC AGCGGCTTCG GCGACGGCTA CA - #ACGGCACC 300- CTGCTGGTGC AGCTCAGCAG CGCCGGAACC CGTACCTACC TCAAGAGCTA CG - #AGGAGGAC 360- CTCGAGGGCC GGCGCTTCGA GGTCGCCCTG GACGGCGACC ACACGGGCGA TC - #TTTCCGCC 420# 447 CCGA CGACGGC- (2) INFORMATION FOR SEQ ID NO:34:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 453 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:- CTCGAAGGCA CGGCCGGCAA CGACACGCTT GGCGGCAGCG ACGCCCACGA GA - #CGCTGCTC 60- GGGCTGGACG GCAACGACCG CCTGAACGGC GGCGCCGGCA ACGACATCCT CG - #ACGGCGGC 120- GCCGGGCGCG ACAACCTGAC CGGCGGCGCG GGCGCCGACC TGTTCCGCGT CT - #CCGCGCGC 180- ACCGACAGCT ACCGCACCGA CAGCGCCAGC TTCAACGACC TGATCACCGA CT - #TCGACGCC 240- AGCCAGGACC GCATCGACCT GTCCGCGCTG GGCTTCACCG GGCTGGGCGA CG - #GCTATAAC 300- GGCACCCTGC TGCTGCAGGT CAGCGCCGAC GGCAGCCGCA CCTATCTGAA GA - #GCCTGGAG 360- GCGGATGCCG AGGGGCGGCG TTTCGAGATC GCCCTGGACG GCAACTTCGC CG - #GCCTGCTC 420# 453 TCTT CGAGCGCACC GCC- (2) INFORMATION FOR SEQ ID NO:35:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 459 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:- ATCGAGGGGG ATGCCGGCGA CAACGCCCTG CTCGGTACCT CGGCCGCCGA GA - #CATTGCTC 60- GGCCACGCCG GCAACGACAC GCTCGACGGC GGGGCCGGCG ACGACATCCT GG - #TCGGCGGC 120- GCCGGGCGCG ACAGCCTCAC CGGCGGCGCC GGAGCGGACG TGTTCCGCTT CG - #ACGCGCTG 180- TCCGACAGCC AGCGCAACTA CGACATCGGC GACAACCAGG GCGACCGCAT CG - #CCGACTTC 240- GCGGTGGGCG AAGACAAGCT CGACGTATCG GCGCTGGGCT TCACCGGGCT GG - #GCGACGGC 300- TACAACGGCA CCCTCGCCCT GGTGCTCAAC AGCGCCGGCG ACCGCACCTA CG - #TGAAAAGC 360- TACGAGAACG GCGCCGACGG CTACCGCTTC GAGTTTTCCC TCGACGGCAA CT - #ATCTGGAG 420# 459 ATTT CATCTTCGCC ACGCCCAGC- (2) INFORMATION FOR SEQ ID NO:36:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 396 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:- CTCGAAGGCA GCGCCGGCAA CGACAGCCTG CAGGGCACGG CCGCCGACGA GG - #TGATCCAC 60- GGCGGCGGCG GGCGCGACAC GCTGGCCGGA GGGGCCGGGG CCGACGTGTT CC - #GCTTTAGC 120- GAACTGACCG ACAGCTACCG AGACAGTGCC AGCTATGCCG ATCTGATCAC TG - #ACTTCGAT 180- GCCAGCGAGG ATCGTATCGA CCTGTCCGGC CTCGGCTTCA GCGGTCTGGG CA - #ACGGCTAC 240- GGCGGTACCC TGGCGCTGCA GGTGAACAGC GCCGGTACCC GCACCTACCT GA - #AGAGCTTC 300- GAGACCAACG CCGCCGGCGA GCGTTTCGAG ATCGCCCTGG ACGGCGACCT GT - #CCGCGCTC 360# 396 TCCT CGACGCGCGT ACCGTA- (2) INFORMATION FOR SEQ ID NO:37:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 459 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:- CTGGCGGGCG GCGACGGCAA CGACACGCTT TCCGGCAGCA GCGCGGCCGA GG - #AACTGCTC 60- GGCGGGGTCG GCAACGACAG CCTGGACGGC GGCGCCGGCA ACGACATCCT CG - #ACGGCGGG 120- GCGGGGCGCG ACACCCTGAG TGGCGGCAGC GGCAGCGACA TCTTCCGCTT CG - #GCGGCGCG 180- CTCGACAGCT TCCGCAACTA CGCCAGCGGG ACGAACGGCA CCGACAGCAT CA - #CCGACTTC 240- ACCCCCGGCG AGGATCTGAT CGACCTCTCC GTGCTCGGCT ACACCGGGCT GG - #GCGACGGC 300- TACAACGGTA CCCTGGCGAT AGTGCTGAAC GACGCCGGCA CCAAGACCTA CC - #TGAAAAAC 360- CGCGAGAGCG ACGCCGAAGG CAACCAGTTC GAGATCGCCC TGGAGGGCAA CC - #ACGCCGAC 420# 459 ACTT CATCTTCGCC ACGGCGGCC- (2) INFORMATION FOR SEQ ID NO:38:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 453 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:- CTCGAAGGCA CCGCGGGCAA CGACGTGCTC AGCGGAACGG GTGCCCACGA GC - #TGATTCTC 60- GGCCTGGCCG GCAACGATCG CCTGGACGGT GGCGCCGGCG ACGACACCCT CG - #ACGGCGGC 120- GCGGGGCGCG ATACCCTGAC CGGCGGCGCG GGCGCCGATA CCTTCCGCTT CT - #CTGCCCGC 180- GAGGACAGTC ACCGCACCGA CAGCGCCAGC TTCACCGACC TGATCACCGA CT - #TCGACGCC 240- AGCCAGGACC GCATCGACCT CTCCGCGCTG GGCTTCACCG GTCTGGGCAA CG - #GTTATGAC 300- GGCACCCTGG CGGTGACCAC CGGTTCCGGC GGCACCCGCA CCTACCTGAA GA - #GCTACGAG 360- GTGGACGCCC AGGGCCGGCG TTTCGAAATC GCCCTGGACG GCAACTTCGT CG - #GCCAGTTC 420# 453 TGTT CGACGCCGCT CCG- (2) INFORMATION FOR SEQ ID NO:39:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 459 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:- GTCACCGGTA CCGAGGGCAA CGACAACCTG TCCGGCACCG ATGCCGGGGA AA - #CCCTCCTG 60- GGCTACGGCG GCAACGACAC CCTCAACGGC GGGGCCGGCA ACGACATCCT GG - #TCGGCGGC 120- GCCGGGCGCG ACACCCTGAC CGGCGGCGCC GGGGCGGACG TGTTCCGCTT CG - #AGGCGCTG 180- TCCGACAGCC AGCGCAACTA CACCGCCGGC GACAACCAGG GCGATTACAT CA - #TCGACTTC 240- GCCGTGGGCG AAGACAGGAT CGACGTATCG GCGCTGGGTT ACACCGGGCT GG - #GCAACGGC 300- CGCAACGGCA CCCTCGCCGT GGTGCTCAAC AGCGCCGGCG ACCGCACCTA CG - #TGAAGAGC 360- TACGACACTG ACGCCAACGG CTATAACTTC GAGCTTTCCC TCGCGGGCAA CT - #ACCAGGGG 420# 459 AGTT CGTCTTCGCC ACGCCCCCG- (2) INFORMATION FOR SEQ ID NO:40:- (i) SEQUENCE CHARACTERISTICS:#pairs (A) LENGTH: 450 base (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: DNA (genomic)- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:- ATCGAGGGAA CCGACGGCAA CGACAGCTTG CAAGGGACCG GGGCCGACGA AC - #TGCTCCTC 60- GGTCTGGGCG GCCGGGACAG CCTGAACGGC GGCGCCGGCG ACGATGTCCT GG - #ATGGCGGG 120- GCGGAGCGCG ACACCCTGAC CGGCGGCACG GGGGCCGACA CCTTCCTGTT CT - #CCGCGCGT 180- ACCGACAGCT ACCGCACCGA CAGCGCCAGC TTCACCGACC TGATCACCGA CT - #TCGATCCC 240- GCCCAGGATC GCATCGACCT GTCCGGCCTG GGCTTCAGCG GTTTCGGCAA CG - #GCTACGAC 300- GGCACCCTGC TGCTGCAGGT CAACGCCGCG GGCACCCGCA CCTACCTGAA GA - #GCCTCGAG 360- GCCGATGCCG ACGGCCAGCG CTTCGAGATC GCCCTGGACG GCGACTTCAG CG - #GCCAGTTG 420# 450 TCTT CGAGGCCGGC- (2) INFORMATION FOR SEQ ID NO:41:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 142 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:- Leu Val Gly Gly Asp Thr Asp Asp Gln Leu Gl - #n Gly Gly Ser Gly Ala# 15- Asp Arg Leu Asp Gly Gly Ala Gly Asp Asp Il - #e Leu Asp Gly Gly Ala# 30- Gly Arg Asp Arg Leu Ser Gly Gly Ala Gly Al - #a Asp Thr Phe Val Phe# 45- Ser Ala Arg Glu Asp Ser Tyr Arg Thr Asp Th - #r Ala Val Phe Asn Asp# 60- Leu Ile Leu Asp Phe Glu Ala Ser Glu Asp Ar - #g Ile Asp Leu Ser Ala#80- Leu Gly Phe Ser Gly Leu Gly Asp Gly Tyr Gl - #y Gly Thr Leu Leu Leu# 95- Lys Thr Asn Ala Glu Gly Thr Arg Thr Tyr Le - #u Lys Ser Phe Glu Ala# 110- Asp Ala Glu Gly Arg Arg Phe Glu Val Ala Le - #u Asp Gly Asp His Thr# 125- Gly Asp Leu Ser Ala Ala Asn Val Val Phe Al - #a Ala Thr Gly# 140- (2) INFORMATION FOR SEQ ID NO:42:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 151 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:- Leu Glu Gly Ser Ala Gly Asn Asp Ala Leu Se - #r Gly Thr Glu Ala His# 15- Glu Thr Leu Leu Gly Gln Ala Gly Asp Asp Ar - #g Leu Asn Gly Asp Ala# 30- Gly Asn Asp Ile Leu Asp Gly Gly Ala Gly Ar - #g Asp Asn Leu Thr Gly# 45- Gly Ala Gly Ala Asp Thr Phe Arg Phe Ser Al - #a Arg Thr Asp Ser Tyr# 60- Arg Thr Asp Ser Ala Ser Phe Asn Asp Leu Il - #e Thr Asp Phe Asp Ala#80- Asp Glu Asp Ser Ile Asp Leu Ser Ala Leu Gl - #y Phe Thr Gly Leu Gly# 95- Asp Gly Tyr Asn Gly Thr Leu Leu Leu Lys Th - #r Asn Ala Glu Gly Thr# 110- Arg Thr Tyr Leu Lys Ser Tyr Glu Ala Asp Al - #a Gln Gly Arg Arg Phe# 125- Glu Ile Ala Leu Asp Gly Asn Phe Thr Gly Le - #u Phe Asn Asp Asn Asn# 140- Leu Leu Phe Asp Ala Ala Pro145 1 - #50- (2) INFORMATION FOR SEQ ID NO:43:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 153 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:- Ala Thr Gly Thr Glu Gly Ser Asp Asn Leu Le - #u Gly Thr Asp Ala Gly# 15- Glu Thr Leu Leu Gly Tyr Gly Gly Asn Asp Th - #r Leu Asn Gly Gly Ala# 30- Gly Asp Asp Ile Leu Val Gly Gly Ala Gly Ar - #g Asp Ser Leu Thr Gly# 45- Gly Ala Gly Ala Asp Val Phe Arg Phe Asp Al - #a Leu Ser Asp Ser Gln# 60- Arg Asn Tyr Thr Thr Gly Asp Asn Gln Ala As - #p Arg Ile Leu Asp Phe#80- Asp Pro Thr Leu Asp Arg Ile Asp Val Ser Al - #a Leu Gly Phe Thr Gly# 95- Leu Gly Asn Gly Arg Asn Gly Thr Leu Ala Va - #l Val Leu Asn Ser Ala# 110- Gly Asp Arg Thr Asp Leu Lys Ser Tyr Asp Th - #r Asp Ala Asn Gly Tyr# 125- Ser Phe Glu Leu Ser Leu Ala Gly Asn Tyr Gl - #n Gly Gln Leu Ser Ala# 140- Glu Gln Phe Val Phe Ala Thr Ser Gln145 1 - #50- (2) INFORMATION FOR SEQ ID NO:44:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 150 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:- Ile Glu Gly Thr Asp Gly Asn Asp Thr Leu Gl - #n Gly Thr Glu Ala Asn# 15- Glu Arg Leu Leu Gly Leu Asp Gly Arg Asp As - #n Leu Asn Gly Gly Ala# 30- Gly Asp Asp Ile Leu Asp Gly Gly Ala Gly Ar - #g Asp Thr Leu Thr Gly# 45- Gly Thr Gly Ala Asp Thr Phe Leu Phe Ser Th - #r Arg Thr Asp Ser Tyr# 60- Arg Thr Asp Ser Ala Ser Phe Asn Asp Leu Il - #e Thr Asp Phe Asp Pro#80- Thr Gln Asp Arg Ile Asp Leu Ser Gly Leu Gl - #y Phe Ser Gly Phe Gly# 95- Asn Gly Tyr Asp Gly Thr Leu Leu Leu Gln Va - #l Asn Ala Ala Gly Thr# 110- Arg Thr Tyr Leu Lys Ser Phe Glu Ala Asp Al - #a Asn Gly Gln Arg Phe# 125- Glu Ile Ala Leu Asp Gly Asp Phe Ser Gly Gl - #n Leu Asp Ser Gly Asn# 140- Val Ile Phe Glu Pro Ala145 1 - #50- (2) INFORMATION FOR SEQ ID NO:45:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 149 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:- Gly Thr Asp Gly Asn Asp Val Leu Val Gly Se - #r Asp Ala Asn Asp Gln# 15- Leu Tyr Gly Gly Ala Gly Asp Asp Arg Leu As - #p Gly Gly Ala Gly Asp# 30- Asp Leu Leu Asp Gly Gly Ala Gly Arg Asp As - #p Leu Thr Gly Gly Thr# 45- Gly Ala Asp Thr Phe Val Phe Ala Ala Arg Th - #r Asp Ser Tyr Arg Thr# 60- Asp Ala Gly Val Phe Asn Asp Leu Ile Leu As - #p Phe Asp Ala Ser Glu#80- Asp Arg Ile Asp Leu Ser Ala Leu Gly Phe Se - #r Gly Phe Gly Asp Gly# 95- Tyr Asn Gly Thr Leu Leu Val Gln Leu Ser Se - #r Ala Gly Thr Arg Thr# 110- Tyr Leu Lys Ser Tyr Glu Glu Asp Leu Glu Gl - #y Arg Arg Phe Glu Val# 125- Ala Leu Asp Gly Asp His Thr Gly Asp Leu Se - #r Ala Ala Asn Val Val# 140- Phe Ala Asp Asp Gly145- (2) INFORMATION FOR SEQ ID NO:46:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 151 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:- Leu Glu Gly Thr Ala Gly Asn Asp Thr Leu Gl - #y Gly Ser Asp Ala His# 15- Glu Thr Leu Leu Gly Leu Asp Gly Asn Asp Ar - #g Leu Asn Gly Gly Ala# 30- Gly Asn Asp Ile Leu Asp Gly Gly Ala Gly Ar - #g Asp Asn Leu Thr Gly# 45- Gly Ala Gly Ala Asp Leu Phe Arg Val Ser Al - #a Arg Thr Asp Ser Tyr# 60- Arg Thr Asp Ser Ala Ser Phe Asn Asp Leu Il - #e Thr Asp Phe Asp Ala#80- Ser Gln Asp Arg Ile Asp Leu Ser Ala Leu Gl - #y Phe Thr Gly Leu Gly# 95- Asp Gly Tyr Asn Gly Thr Leu Leu Leu Gln Va - #l Ser Ala Asp Gly Ser# 110- Arg Thr Tyr Leu Lys Ser Leu Glu Ala Asp Al - #a Glu Gly Arg Arg Phe# 125- Glu Ile Ala Leu Asp Gly Asn Phe Ala Gly Le - #u Leu Gly Ala Gly Asn# 140- Leu Leu Phe Glu Arg Thr Ala145 1 - #50- (2) INFORMATION FOR SEQ ID NO:47:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 153 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:- Ile Glu Gly Asp Ala Gly Asp Asn Ala Leu Le - #u Gly Thr Ser Ala Ala# 15- Glu Thr Leu Leu Gly His Ala Gly Asn Asp Th - #r Leu Asp Gly Gly Ala# 30- Gly Asp Asp Ile Leu Val Gly Gly Ala Gly Ar - #g Asp Ser Leu Thr Gly# 45- Gly Ala Gly Ala Asp Val Phe Arg Phe Asp Al - #a Leu Ser Asp Ser Gln# 60- Arg Asn Tyr Asp Ile Gly Asp Asn Gln Gly As - #p Arg Ile Ala Asp Phe#80- Ala Val Gly Glu Asp Lys Leu Asp Val Ser Al - #a Leu Gly Phe Thr Gly# 95- Leu Gly Asp Gly Tyr Asn Gly Thr Leu Ala Le - #u Val Leu Asn Ser Ala# 110- Gly Asp Arg Thr Tyr Val Lys Ser Tyr Glu As - #n Gly Ala Asp Gly Tyr# 125- Arg Phe Glu Phe Ser Leu Asp Gly Asn Tyr Le - #u Glu Leu Leu Gly Asn# 140- Glu Asp Phe Ile Phe Ala Thr Pro Ser145 1 - #50- (2) INFORMATION FOR SEQ ID NO:48:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 132 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:- Leu Glu Gly Ser Ala Gly Asn Asp Ser Leu Gl - #n Gly Thr Ala Ala Asp# 15- Glu Val Ile His Gly Gly Gly Gly Arg Asp Th - #r Leu Ala Gly Gly Ala# 30- Gly Ala Asp Val Phe Arg Phe Ser Glu Leu Th - #r Asp Ser Tyr Arg Asp# 45- Ser Ala Ser Tyr Ala Asp Leu Ile Thr Asp Ph - #e Asp Ala Ser Glu Asp# 60- Arg Ile Asp Leu Ser Gly Leu Gly Phe Ser Gl - #y Leu Gly Asn Gly Tyr#80- Gly Gly Thr Leu Ala Leu Gln Val Asn Ser Al - #a Gly Thr Arg Thr Tyr# 95- Leu Lys Ser Phe Glu Thr Asn Ala Ala Gly Gl - #u Arg Phe Glu Ile Ala# 110- Leu Asp Gly Asp Leu Ser Ala Leu Gly Gly Al - #a Asn Leu Ile Leu Asp# 125- Ala Arg Thr Val 130- (2) INFORMATION FOR SEQ ID NO:49:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 153 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:- Leu Ala Gly Gly Asp Gly Asn Asp Thr Leu Se - #r Gly Ser Ser Ala Ala# 15- Glu Glu Leu Leu Gly Gly Val Gly Asn Asp Se - #r Leu Asp Gly Gly Ala# 30- Gly Asn Asp Ile Leu Asp Gly Gly Ala Gly Ar - #g Asp Thr Leu Ser Gly# 45- Gly Ser Gly Ser Asp Ile Phe Arg Phe Gly Gl - #y Ala Leu Asp Ser Phe# 60- Arg Asn Tyr Ala Ser Gly Thr Asn Gly Thr As - #p Ser Ile Thr Asp Phe#80- Thr Pro Gly Glu Asp Leu Ile Asp Leu Ser Va - #l Leu Gly Tyr Thr Gly# 95- Leu Gly Asp Gly Tyr Asn Gly Thr Leu Ala Il - #e Val Leu Asn Asp Ala# 110- Gly Thr Lys Thr Tyr Leu Lys Asn Arg Glu Se - #r Asp Ala Glu Gly Asn# 125- Gln Phe Glu Ile Ala Leu Glu Gly Asn His Al - #a Asp Gln Leu Asp Ala# 140- Ser Asp Phe Ile Phe Ala Thr Ala Ala145 1 - #50- (2) INFORMATION FOR SEQ ID NO:50:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 151 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:- Leu Glu Gly Thr Ala Gly Asn Asp Val Leu Se - #r Gly Thr Gly Ala His# 15- Glu Leu Ile Leu Gly Leu Ala Gly Asn Asp Ar - #g Leu Asp Gly Gly Ala# 30- Gly Asp Asp Thr Leu Asp Gly Gly Ala Gly Ar - #g Asp Thr Leu Thr Gly# 45- Gly Ala Gly Ala Asp Thr Phe Arg Phe Ser Al - #a Arg Glu Asp Ser His# 60- Arg Thr Asp Ser Ala Ser Phe Thr Asp Leu Il - #e Thr Asp Phe Asp Ala#80- Ser Gln Asp Arg Ile Asp Leu Ser Ala Leu Gl - #y Phe Thr Gly Leu Gly# 95- Asn Gly Tyr Asp Gly Thr Leu Ala Val Thr Th - #r Gly Ser Gly Gly Thr# 110- Arg Thr Tyr Leu Lys Ser Tyr Glu Val Asp Al - #a Gln Gly Arg Arg Phe# 125- Glu Ile Ala Leu Asp Gly Asn Phe Val Gly Gl - #n Phe Asn Asp Gly Asn# 140- Leu Leu Phe Asp Ala Ala Pro145 1 - #50- (2) INFORMATION FOR SEQ ID NO:51:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 153 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:- Val Thr Gly Thr Glu Gly Asn Asp Asn Leu Se - #r Gly Thr Asp Ala Gly# 15- Glu Thr Leu Leu Gly Tyr Gly Gly Asn Asp Th - #r Leu Asn Gly Gly Ala# 30- Gly Asn Asp Ile Leu Val Gly Gly Ala Gly Ar - #g Asp Thr Leu Thr Gly# 45- Gly Ala Gly Ala Asp Val Phe Arg Phe Glu Al - #a Leu Ser Asp Ser Gln# 60- Arg Asn Tyr Thr Ala Gly Asp Asn Gln Gly As - #p Tyr Ile Ile Asp Phe#80- Ala Val Gly Glu Asp Arg Ile Asp Val Ser Al - #a Leu Gly Tyr Thr Gly# 95- Leu Gly Asn Gly Arg Asn Gly Thr Leu Ala Va - #l Val Leu Asn Ser Ala# 110- Gly Asp Arg Thr Tyr Val Lys Ser Tyr Asp Th - #r Asp Ala Asn Gly Tyr# 125- Asn Phe Glu Leu Ser Leu Ala Gly Asn Tyr Gl - #n Gly Leu Leu Gly Ala# 140- Glu Gln Phe Val Phe Ala Thr Pro Pro145 1 - #50- (2) INFORMATION FOR SEQ ID NO:52:- (i) SEQUENCE CHARACTERISTICS:#acids (A) LENGTH: 150 amino (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear- (ii) MOLECULE TYPE: protein- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:- Ile Glu Gly Thr Asp Gly Asn Asp Ser Leu Gl - #n Gly Thr Gly Ala Asp# 15- Glu Leu Leu Leu Gly Leu Gly Gly Arg Asp Se - #r Leu Asn Gly Gly Ala# 30- Gly Asp Asp Val Leu Asp Gly Gly Ala Glu Ar - #g Asp Thr Leu Thr Gly# 45- Gly Thr Gly Ala Asp Thr Phe Leu Phe Ser Al - #a Arg Thr Asp Ser Tyr# 60- Arg Thr Asp Ser Ala Ser Phe Thr Asp Leu Il - #e Thr Asp Phe Asp Pro#80- Ala Gln Asp Arg Ile Asp Leu Ser Gly Leu Gl - #y Phe Ser Gly Phe Gly# 95- Asn Gly Tyr Asp Gly Thr Leu Leu Leu Gln Va - #l Asn Ala Ala Gly Thr# 110- Arg Thr Tyr Leu Lys Ser Leu Glu Ala Asp Al - #a Asp Gly Gln Arg Phe# 125- Glu Ile Ala Leu Asp Gly Asp Phe Ser Gly Gl - #n Leu Asp Ser Gly Asn# 140- Val Ile Phe Glu Ala Gly145 1 - #50__________________________________________________________________________

Claims

1. A DNA compound comprising a sequence encoding mannuronan C- 5-epimerase, said sequence comprising one or more sequences selected from the group consisting of a DNA A block comprising the DNA of SEQ ID NO:7 or any one of SEQ ID NOS:17-22, or a DNA molecule encoding an amino acid sequences comprising an amino acid sequence of SEQ ID NO:8 or any one of SEQ ID NOS:23-28, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:7 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C.; and a DNA R block comprising the DNA of SEQ ID NO:9 or any one of SEQ ID NOS:29-40, or a DNA molecule encoding an amino sequence comprising an amino acid sequence of SEQ ID NO:10 or any one of SEQ ID NOS:41-52, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:9 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C.; and being isolated from a natural source or being derived synthetically.

2. An isolated DNA compound according to claim 1 encoding mannuronan C-5-epimerase that comprises at least one DNA block A comprising the DNA of SEQ ID NO:7 or any one of SEQ ID NOS:17-22, and a DNA block R comprising the DNA of SEQ ID NO:9 or any one of SEQ ID NOS:29-40 in a number of 0 to 5.

3. An isolated DNA compound comprising at least a portion of the DNA sequence as set forth in SEQ ID NO:1 wherein said compound comprises one or more sequences encoding a DNA A block comprising the DNA of SEQ ID NO:7 or any one of SEQ ID NOS:17-22, or a DNA molecule encoding an amino acid sequence comprising an amino acid sequence of SEQ ID NO:8 or any one of SEQ ID NOS:23-28, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:7 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C.; and/or a DNA R block comprising the DNA of SEQ ID NO:9 or any one of SEQ ID NOS:29-40, or a DNA molecule encoding an amino sequence comprising an amino acid sequence of SEQ ID NO:10 or any one of SEQ ID NOS:41-52, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:9 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C.

4. An isolated DNA compound according to claim 1 that encodes at least a portion of the protein amino acid sequence as set, said sequence containing SEQ ID NOS:2,3,4 and 5.

5. A recombinant DNA vector that comprises the DNA sequence of claim 1.

6. A method for constructing a recombinant host cell for production of enzymes having mannuronan C-5-epimerase activity and/or the microbiological production of alginates which comprises transforming a host cell with the DNA of claim 1.

7. The method according to claim 6 which comprises the microbiological production of alginates having a high G block content of 75-98%.

8. The method according to claim 6 which comprises the microbiological production of alginates wherein the DNA sequence is selected to produce an alginate having a desired M/G block content.

9. A method for the production of enzymes having mannuronan C-5-epimerase activity comprising constructing a recombinant host cell capable of expressing epimerase activity, characterized by transforming said host cell with a recombinant DNA expression vector that comprises:

(a) a promoter and translational activating sequence that function in said host cell; and

(b) a DNA sequence encoding mannuronan C-5-epimerase comprising a DNA block A comprising the DNA of SEQ ID NO:7 or any one of SEQ ID NOS:17-22, or a DNA molecule encoding an amino acid sequence comprising an amino acid sequence of SEQ ID NO:8 or any one of SEQ ID NOS:23-28, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:7 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C. and/or a DNA block R comprising the DNA of SEQ ID NO:9 or any one of SEQ ID NOS:29-40, or a DNA molecule encoding an amino sequence comprising an amino acid sequence of SEQ ID NO:10 or any one of SEQ ID NOS:41-52, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:9 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C., positioned for expression from said promoter and translational activity sequence.

10. A process for the bacterial production of alginates comprising selectively inactivating one or more DNA sequences containing at least one A block comprising the DNA of SEQ ID NO:7 or any one of SEQ ID NOS:17-22, or a DNA molecule encoding an amino acid sequence comprising an amino acid sequence of SEQ ID NO:8 or any one of SEQ ID NOS:23-28, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:7 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C., or R block comprising the DNA of SEQ ID NO:9 or any one of SEQ ID NOS:29-40, a a DNA molecule encoding an amino sequence comprising an amino acid sequence of SEQ ID NO:10 or any one of SEQ ID NOS:41-52, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:9 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C. and encoding mannuronan C-5-epimerases in a natural host cell wherein said mannuronan C-5-epimerases produce alginates.

11. Process according to claim 10 for the production of pure poly-M alginate or alginates having a G block content from 0 to 25%.

12. Process according to claim 10 for the production of alginates wherein said DNA sequences are selectively inactivated to produce alginates having a desired M/G block content and G block distribution.

13. A DNA compound according to claim 1 wherein said compound is not naturally occurring in an alginate producing host and comprises at least one A block comprising the DNA of SEQ ID NO:7 or any one of SEQ ID NOS:17-22 and at least one R block comprising the DNA of SEQ ID NO:9 or any one of SEQ ID NOS:29-40 isolated from a natural source or being derived synthetically.

14. A DNA compound encoding a mannuronan C-5-epimerase, wherein said C-5-epimerase comprises an amino acid sequence corresponding to that of a mannuronan C-5-epimerase produced by Azotobacter vinelandii.

15. A DNA compound comprising a sequence encoding mannuronan C-5-epimerase, said sequence comprising one or more sequences of a DNA A block comprising the DNA of SEQ ID NO:7 or any one of SEQ ID NOS:17-22, or a DNA molecule encoding an amino acid sequence comprising an amino acid sequence of SEQ ID NO:8 or any one of SEQ ID NOS:23-28, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:7 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C. and being isolated from a natural source or being derived synthetically.

16. The DNA compound of claim 15 further comprising one or more sequences of a DNA R block comprising the DNA of SEQ ID NO:9 or any one of SEQ ID NOS:29-40, or a DNA molecule encoding an amino sequence comprising an amino acid sequence of SEQ ID NO:10 or any one of SEQ ID NOS:41-52, or a DNA molecule which hybridizes to consensus sequence SEQ ID NO:9 under high stringency conditions comprising a wash step of 3.2 M tetramethylammonium chloride at 50.degree. C.