WO1998058065A1

WO1998058065A1 - Mold-resistant plants and method of construction of same

Info

Publication number: WO1998058065A1
Application number: PCT/JP1998/002719
Authority: WO
Inventors: Makoto Kakitani; Naoyuki Umemoto; Masayoshi Tsukahara; Isao Ishida
Original assignee: Kirin Beer Kabushiki Kaisha
Priority date: 1997-06-18
Filing date: 1998-06-18
Publication date: 1998-12-23
Also published as: CN1266457A; CA2294652A1; AU8035598A

Abstract

DNAs encoding chimeric proteins each having a domain containing at least the elicitor-binding site of an elicitor receptor and another domain containing at least the signal transduction motif of a protein capable of imparting disease resistance to plants; recombinant vectors containing these DNAs; transformed plants prepared by using these vectors or offsprings thereof; and a method for the construction of disease resistant plants characterized by transferring the above vectors into plant hosts.

Description

Specification

Mold-resistant plant and method for producing the same

The present invention relates to a DNA encoding a protein that causes plant disease resistance, a plant disease-resistant plant, and a method for producing the same. More specifically, a domain containing at least an elicitor binding site of an elicitor receptor, and a plant-derived plant The present invention relates to a DNA encoding a chimeric protein (a chimeric receptor 1) containing a domain containing a signal transmission motif of a resistance gene, a plant disease-resistant plant into which the DNA has been introduced, and a method for producing the same. Background art

It is known that plants, when infected with a pathogenic bacterium, exhibit a resistance reaction in which a new antibacterial substance is synthesized and accumulated in the plant [M. Yoshikawa (1978) Nature 257: 546]. Substances that induce such a plant resistance response have been found in the plant fungus [N. T. Keen (1975) Science 187: 74], and these substances are collectively referred to as elicitors.

The elicitor includes degraded products produced by a hydrolase derived from a plant or a power plant, a protein elicitor produced by a power plant, and an AVR gene produced by a power plant.

Various products such as (avirulence gene) products are known. For example, polygalacturonic acid produced by polygalacturonase as the plant-derived degradation product, and chitin oligosaccharide produced by chitinase as a mold-derived degradation product

(N-acetyl chitooligosaccharide) or glucan produced by dalcanase. Examples of the proteinaceous elicitor include mold-derived elicitin belonging to the genus Phytophthora. Examples of the gene product include Avr4 and Avr9 derived from Cladosporium fulvum, and nipl derived from Lincosporium 'Secaris (Rhynchosporium secalis). These elicitors interact specifically with plant resistance gene products, including the elicitor receptor on the plant cell membrane, to produce An intrusion is recognized and a signal is issued. Then, various signals are transmitted to the nucleus to prevent mold from invading the plant, namely, cell death, called hypersensitivity, production of hydrogen peroxide, expression of PR protein, Causes expression of lipoxygenase, production of a resistant substance called huitarexin, etc. (W. Knogge (1996) Plant Cell 8: 1711; AF Bent (1996) Plant Cell 8: 1757; KEHa dandelion ond-Kosack and JDG Jones (1996) Plant Cell 8: 1773).

The biochemical processes from infection by plant pathogens to resistance are thought to be as follows, taking the synthesis and accumulation of phytoalexin as an example.

First, when hyphae of a pathogenic bacterium enter a plant cell, dalkanase present in the plant cell works to cut the polysaccharide on the cell wall surface of the pathogenic bacterium and release elicitor. Elysita is a polysaccharide made from glucose and is a 1,6-linked and | 3-1,3-linked glucan [JK Sharp et al. (1984) J. Biol. Chem. 259: 11321, M. Yoshikawa (1990) Plant Cell Engineering 1.2: 695, Y. Okinaka et al. (1995) Plant Physiol. 109: 839]. When this elicitor binds to a receptor present in plant cells, it produces a second messenger that plays a role in signal transduction, and this signaling substance enters the nucleus of the plant cell and transcribes the gene encoding the phytoalexin synthase. Activates phytoalexin synthase. At the same time, the phytolexin degradation system is suppressed, and phytolexin is efficiently accumulated. For example, when the plant is soybean, phytoalexin, which plays an important role in soybean resistance, is called glyceroline, and its structure has been determined [M. Yoshikawa et al. (1978) Physiol. Pathol. 12: 73] ₀

Therefore, a specific receptor (ER) for dalcan elicitor from Phytophthora megasperaaf. Sp. Glycinea, a kind of soybean pathogenic fungi, plays an important role in the synthesis and accumulation of the antibacterial glyceolin. It is thought to be a receptor protein that plays This specific receptor for glucan elicitor was separated and purified from the cell membrane fraction of soybean root by Kakitani et al. (JP-A-06-321995). Umemoto et al. [N. Umemoto et al. (1997) Pro Natl. Acad. Sci. Loaned.

In recent years, many cloning of plant-derived resistance genes have been reported, including characteristic motifs or domains involved in signal transduction, such as leucine-rich repeats, leucine zippers, nucleic acid binding sites ( Some or all of the P-loop and serine / threonine-kinase domains have been found. Plant-derived disease resistance genes that are expected to contain motifs or domains involved in these signal transductions include, for example, the Arabidopsis resistance gene RPS2 [M. Mindrinos et al. (1994) Cell 78: 1089, AF Bent et al. (1994) Science 265: 1856], Arabidopsis resistance gene RPM1 [MR Grant et al. (1995) Science 269: 843], tobacco resistance gene N [S. Whitham et al. ( 1994) Cell 78: 1101], flax-derived resistance gene L6 [GJ Lawrence et al. (1995) Plant Cell 7: 1195], tomato-derived resistance gene Pto [GB Martin et al. (1993) Science 262: 1432], Prf, a resistance gene derived from tomato [JM Salmeron et al. (1996) Cell 86: 123], Cf-9, a resistance gene derived from tomato [D. to Jones et al. (1994) Science 266: 789] ), The resistance gene Cf-2 from tomato [. S. Dixon et al. (1996) Cell 84: 451], the resistance gene Xa21 from rice (W. -Y. Song et al. (1995) Science 270: 1804].

On the other hand, in contrast to plant cells, numerous reports have been made on cloning of receptors derived from animal cells. In addition, many studies have been conducted on quinula receptors composed of extracellular and intracellular domains derived from different types of receptors. For example, an extracellular domain that recognizes a ligand of one receptor is replaced with an extracellular domain of a receptor that recognizes another ligand, and the replaced extracellular domain and an intracellular cell that transmits a signal to the nucleus. It has been reported that when a chimeric receptor fused with a domain is expressed in a cell, the chimeric receptor recognizes a ligand recognized by the newly replaced extracellular domain and transduces a signal inside the cell (H. Riedel et al. (1986) Nature 324: 68, J. Lee et al. (1989) EMB0 J. 8: 167, H. Lehvaslaiho et al. (1989) EMB0 J. 8: 159, S. Lev et al. (1990) Mol. Cell. Biol. 10: 6064] has been. Furthermore, the same function can be achieved by introducing and expressing the gene encoding the receptor of one animal cell into another type of animal cell.For example, the gene can be expanded by expressing the human-derived GM-CSF receptor in mouse 3T3 cells. It has been reported that facilitating signal transduction is observed [S. Watanabe et al. (1993) MoI. Cell. Biol. 13: 1440].

However, as far as the present inventors know, there have been no reports on chimeric receptors in plants, and no reports suggest that such quinula receptors function in plants. Disclosure of the invention

The aforementioned dalcan elicitor receptor lacks characteristic motifs or domains involved in signal transduction, such as those found in plant-derived resistance genes, and may lack signal transduction domains It has been suggested. In addition, from the study of chimeric receptors in animal cell-derived receptors, the present inventors have found that even in plant cells, a domain that recognizes a certain ligand is involved in signal transduction derived from another protein (including a receptor) in a domain that recognizes a certain ligand. Is it possible that signal transduction is carried out by recognizing the ligand even if a quinula receptor linked to a domain is introduced and expressed? I wondered if signaling could occur even if they were not of the same plant species. That is, a gene encoding a chimeric receptor in which an elicitor receptor is linked to an intracellular signaling domain of a plant-derived receptor, or an elicitor receptor and a plant-derived disease resistance gene (for example, Arabidopsis-derived Cells encoding the resistance gene RPS2, Arabidopsis resistance gene RPM1, tobacco resistance gene N, flax resistance gene L6, tomato resistance gene Pto, rice resistance gene Xa21, etc. Synthesizes a gene encoding a chimeric receptor linked to a domain predicted to be involved in intracellular signal transduction, and if it can be expressed in plants regardless of plant type, expresses only the elicitor receptor We thought that the efficiency of signal transmission might be higher than in the case. The result is a distinctly stronger resistance to pathogenic mold It is possible to produce plants with sexual properties, improve the productivity of agricultural crops, and reduce the economic effect and environmental pollution due to the reduction of pesticide use.

The present invention relates to a DNA encoding a chimeric protein comprising a domain containing at least an elicitor-binding site of an elicitor receptor and a domain containing a signaling motif of a plant-derived resistance gene, and a plant into which the DNA has been introduced. It is intended to provide a disease resistant plant and a method for producing the same.

The present inventors have made intensive efforts to solve the above problems, and as a result, a domain predicted to be involved in signal transduction encoded by a plant-derived resistance gene, and a domain including at least an elicitor binding site of an elicitor receptor. By synthesizing a gene encoding the chimeric receptor of the present invention and introducing it into Arabidopsis thaliana and tobacco plants and expressing it, the plant succeeded in imparting disease resistance to the plant and completed the present invention. It led to.

That is, the present invention is a DNA encoding a chimeric protein containing at least a domain containing an elicitor-binding site of an elicitor-receptor and at least a domain containing a signal transmission motif of a protein conferring plant disease resistance. This chimeric protein functions as a receptor and brings disease resistance to plants.

Examples of elicitors include glucan, polygalacturonic acid, N-acetyl chitooligosaccharide, elicitin, Cladosporium. Fulbam-derived AVr gene product or lincosporium. Secalis-derived nip 1 gene product. Are their receptors. Further, as the elicitor binding site of the glucan elicitor receptor, a protein containing at least the amino acid sequence represented by SEQ ID NO: 27, or at least one amino acid in the amino acid sequence is deleted, substituted, added or inserted. And a protein having the amino acid sequence of the present invention and having the ability to bind to Dalkan elicitor. Further, examples of the domain containing the signal transduction motif include those containing at least one selected from the group consisting of leucine-rich repeat, leucine zipper, nucleic acid binding site, and serine threonine kinase domain. Derived Pt0 gene, Prf gene, Cf-2 gene or Cf-19 gene Signaling domain of the expression product of the Xa21 gene from rice, the signaling domain of the expression product of the RPS2 gene or the RPM1 gene from the Arabidopsis thaliana (for example, The signaling domain of the expression product of the L6 gene derived from flax or the signaling domain of the expression product of the N gene derived from tobacco.

Further, the present invention includes a chimeric protein comprising the amino acid sequence represented by SEQ ID NO: 5, 7 or 9, or an amino acid sequence wherein at least one amino acid is deleted, substituted, added or inserted in the amino acid sequence. DNA encoding a chimeric protein that produces plant disease resistance. Specific examples of the DNA include those containing the nucleotide sequence represented by SEQ ID NO: 6, 8 or 10.

Furthermore, the present invention is a recombinant vector containing the DNA.

Furthermore, the present invention is a transformed plant transformed by the recombinant vector or a progeny thereof.

Furthermore, the present invention relates to a plant having plant disease resistance into which the DNA is introduced and expressing the plant, or a progeny thereof.

Furthermore, the present invention is a method for producing a plant disease-resistant plant, which comprises introducing the recombinant vector into a plant.

Furthermore, the present invention is a method for producing a plant having plant disease resistance or progeny thereof by integrating the DNA into a plant chromosome and expressing the DNA. Hereinafter, the present invention will be described in detail.

The DNA of the present invention is obtained by linking a DNA encoding at least a domain containing at least an elicitor binding site of an elicitor receptor and a DNA encoding a domain containing at least a signal transduction motif of a protein that causes plant disease resistance. It is also referred to as DNA or chimeric DNA encoding a chimeric receptor.

The elicitor receptor includes elicitor of a plant degradation product such as polygalacturonic acid, or a receptor that binds to elicitor of a fungal cell wall degradation product such as dalcan or N-acetylchitooligosaccharide, Mold like elicitin A receptor for the proteinaceous elicitor to be produced, or an elicitor receptor that binds to an elicitor such as an AVr gene product derived from Cladosporidium flubum, a nip1 gene product derived from Lincosporium secalis, and the like. Methods for cloning cDNAs (hereinafter referred to as “ERDNA”) encoding these elicitor receptors include the following.

That is, the above-mentioned elicitor is labeled with an isotope, and the binding activity to the labeled elicitor is measured from the plant cell membrane fraction to purify the elicitor receptor which specifically binds to elicitor. PCR primers are designed from the partial amino acid sequence information obtained from the purified elicitor receptor, and the cDNA library is used for the purpose from a cDNA library using a probe obtained from a PCR derived from plant cell-derived DNA. C Clean the DNA.

On the other hand, the DNA encoding the signal transduction motif in the present invention is known as described above, and can be cloned using PCR or the like.

Next, ligation of the ERDN A with the DNA encoding the signaling motif is performed. The above two DNAs can be ligated by digesting these DNAs with an appropriate restriction enzyme and then ligating them using a ligase, or by combining ERDN A with a DNA encoding a signal transduction motif with an appropriate oligonucleotide. It is done by combining. Alternatively, the ligation may be performed by inserting the above DNA into an appropriate plasmid. The position before and after the position where the ERDNA and the DNA encoding the signal transduction motif are linked is not particularly limited, and even if the ERDNA is located upstream and the DNA encoding the signaling motif is located downstream. , And vice versa.

Finally, the base sequence of the chimeric DNA is determined by a known method (didoxy method, Maxam-Gilbert method, etc.). Usually, the base sequence is determined using a commercially available automatic base sequencer.

FIG. 1 shows a schematic diagram of a chimeric DNA of the present invention, taking as an example a chimeric DNA of a DNA encoding a dalcan elicitor receptor (ER) and an RPS2 gene derived from Arabidopsis thaliana. In FIG. 1, CER is a chimeric DNA consisting of an approximately one-third region (SEQ ID NO: 29) on the 5 ′ side of the RPS2 gene and the entire ERDNA (SEQ ID NO: 2). Cut part and ERDN This is the one in which A is located upstream and the RPS2 gene is located downstream (the arrangement in the CER has been replaced).

The elicitor binding site of the dalcan elicitor receptor corresponds to the 239th to 442nd amino acid sequence of the amino acid sequence represented by SEQ ID NO: 1, and has the amino acid sequence represented by SEQ ID NO: 27. Here, a protein comprising at least the amino acid sequence represented by SEQ ID NO: 27, or an amino acid sequence in which at least one amino acid is deleted, substituted, added or inserted in the amino acid sequence, and has a binding ability to glucan elicitor. The length of the elicitor-receptor is not limited as long as it is a protein having the following structure: it may be a protein having an appropriate truncated form (see “2-11” and “2-16” in FIG. 1). , 1-5). The DNA encoding the amino acid sequence of the elicitor-binding site represented by SEQ ID NO: 27 includes a DNA represented by SEQ ID NO: 28.

The nucleotide sequences of these chimeric DNAs and the amino acid sequences of the chimeric proteins encoded by the DNAs are shown in SEQ ID NOS: 5 and 6 for CER, SEQ ID NOs: 7 and 8 for IER, and The ER is shown in SEQ ID NOs: 9 and 10, and as long as these chimeric proteins have a binding function as an elicitor receptor and bring about plant disease resistance activity, the amino acid sequence (SEQ ID NOs: 5, 7, and 10) In 9), it means that at least one amino acid may cause mutation such as deletion, substitution, addition, or insertion. The DNA of the present invention also includes, in addition to the nucleotide sequence encoding an amino acid contained in the chimeric protein of the present invention, DNA encoding the same polypeptide that differs only in degenerate codons (referred to as degenerate isomers). It is. For example, the codon (AAC) corresponding to Asn and the codon (eg, AAT) that has degenerate relation to it are called degenerate isomers.

In addition, introduction of the above-mentioned mutation is described in, for example, DFMark et al., Proc. Natl. Acad. Sci. USA, Vol. 81, p5662-5666, 1984, PCT W085 / 00817, published on February 28, 1985, RP Wharton et al. Nature, Vol. 316, p60 605, Aug. 15, 1985, site-specific mutagenesis, etc., which is a well-known technique before the filing date of the present application. Further, once the above-mentioned nucleotide sequence is determined, it is then subjected to chemical synthesis, PCR or hybridization using a DNA fragment having the nucleotide sequence as a probe, whereby The DNA of the present invention can be obtained.

The DNA sequence encoding the chimeric receptor used in the present invention preferably has at least one stop codon (eg, TAG) adjacent to the 3'-end. Also, if desired, an ATG sequence encoding a methionine for translation initiation along with the translation frame at the 5'-upstream, and other 5'-upstream and 3'-downstream non-translated regions of appropriate length as untranslated regions DNA may bind.

Representative forms of the DNA sequence encoding the chimeric receptor used in the present invention include a plasmid or phage DNA in which this DNA sequence is inserted as a member, and plasmid or phage. Alternatively, the DNA sequence is present in a microorganism (especially a bacterium) or a phage particle or in a plant in which the DNA sequence is inserted into genomic DNA. The bacteria mentioned here include Escherichia coli bacterium. In order to stably express the DNA sequence encoding the chimeric receptor in plants, a promoter, a DNA coding for a translation initiation codon (ATG) and a terminator are added to this DNA sequence in an appropriate combination. It is good.

The recombinant vector of the present invention can be constructed by incorporating a DNA encoding the chimeric receptor of the present invention (hereinafter also referred to as “chimeric DNA”) into an appropriate vector.

The vector for incorporating the chimeric DNA of the present invention is not particularly limited as long as it can replicate in a host, and examples thereof include plasmid DNA and phage DNA. A binary vector, one of the plasmid DNAs, can be prepared from Escherichia coli or Agrobacterium by an alkali extraction method (Birnboim, H. & Doly, J. (1979) Nucleic acid Res 7: 1513) or the like. Examples of the binary one vector include ρΒΙΙΟΙ121, ρ 挙げ and the like. Further, as commercially available plasmids, for example, PUC118 (manufactured by Takara Shuzo), PUCI19 (manufactured by Takara Shuzo), pBluescript SK + (manufactured by Stratagene), pGEM-T (manufactured by Promega) and the like may be used.

Examples of the phage DNA include M13mpl8, M13mpl9, and M13tvl8.

To link a promoter to a vector, first, a purified chimera of the present invention A method is used in which DNA is cleaved with an appropriate restriction enzyme, inserted into an appropriate restriction enzyme site of vector DNA or a multi-cloning site, and ligated to the vector. Here, the chimeric DNA to be expressed needs to be incorporated into a vector so that the function of the DNA is exhibited. Therefore, in addition to the promoter and the above genes, a signal peptide gene, a terminator, a drug resistance gene and the like can be incorporated into the recombinant vector of the present invention. In this case, examples of the signal peptide gene include a tomato (Lycopersicon esculentum) 3-1,3 glucanase-sidinal peptide gene and the like. Examples of the enhancer include a tobacco mosaic virus Ω sequence and the like. Examples include kanamycin resistance gene and hygromycin resistance gene.

Examples of the above promoter include a promoter of a gene encoding a small subunit of ribulose-1,5-2 phosphoric acid carboxylase [R. Fluhr et al. Pro Natl, Acad. Sci. USA (1986) 83: 2358], Promoter of nopaline synthase (NOS) gene [WHR Langridge et al. Plant Cell Rep. (1985) 4: 355], Promoter that produces mosquito reflex mosaic virus 19S-RNA [H. Guilley et al. Cell (1982) 30: 763], and a promoter that produces cauliflower mosaic virus 35S-RNA [JT Odell et al. Nature (1985) 313: 810]. Examples of terminators include the terminator of the nopaline synthase gene [A. Depicker et al. J. Mol. ApI. Gen. (1982) 1: 561], and the terminator of the octopine synthase gene. [J. Gielen et al. EMBO J. (1984) 3: 835] can be used.

Specifically, as shown in FIG. 2, a cauliflower mosaic virus 35S promoter (p35S) is ligated upstream of the chimeric DNA, and a terminator (NosT) of the nopaline synthase gene is ligated downstream (referred to as an ER cassette). In addition, a cassette in which a kanamycin resistance gene (NPT II) is connected between the promoter and terminator of the nopaline synthase gene (NPT II cassette), and a hygromycin resistance gene is connected between p35S and NosT. We also make cassettes (called Hm cassettes). Then, the recombinant vector of the present invention can be obtained by connecting the NPT II cassette upstream of the ER cassette and connecting the Hm cassette downstream of the ER cassette. (Figure 2).

Various studies on elicitors and their receptors suggest that, for example, in the case of glucan elicitors and their receptors, the glucan elicitor receptor is important in plants for resistance to a wide variety of forceps, including dalkanes as cell wall components. It is suggested that it plays an important role. Then, the receptor gene was cloned, and the primary sequence was examined, suggesting that the elicitor receptor lacked a domain involved in signal transduction. On the other hand, the reported plant-derived resistance genes are roughly classified into four classes according to their structures as follows [M. S. Dixon et al. (1996) Cell 84: 451]. Class 1: Contains leucine-rich repeats and nucleic acid binding sites, some also have leucine zippers and are probably localized in the cytoplasm. Resistance genes that meet these conditions include tobacco-derived resistance gene N, Arabidopsis-derived resistance genes RPS2 and RPM1, flax-derived resistance gene L6, and tomato-derived resistance gene Prf.

Class 2: Leucine-ritslipipit, which has a membrane-binding site and is probably located on the cell surface, including tomato-derived resistance genes Cf-2 and Cf-9.

Class 3: Has a serine / threonine kinase domain and includes the tomato-derived resistance gene Pto.

Class 4: a leucine-rich repeat, a transmembrane site, with a serine / threonine-like kinase domain, and possibly a leucine-rich repeat recognizing elicitor, thereby activating a serine / threonine kinase located in the cell. And a rice-derived resistance gene Xa-21.

Thus, plant-derived resistance genes contain one or more motifs or domains predicted to be involved in signal transduction, i.e., leucine-rich repeats, leucine zippers, nucleic acid binding sites, and serine / threonine kinase domains. Yes.

Therefore, in the present invention, at least the elicitor binding of the elicitor receptor The domain containing the site and the codes of plant-derived disease resistance genes (RPS2 gene, RPM1 gene, N gene, L6 gene, Pto gene, Xa21 gene, Cf-2 gene, Cf-9 gene, Prf gene, etc.) A DNA encoding a chimeric protein obtained by linking to a domain involved in signal transduction is produced and expressed.

For example, a chimera of a DNA encoding a domain containing at least the elicitor-binding site of the glucan elicitor receptor and a DNA encoding a domain containing a leucine zipper and a nucleic acid binding site of a resistance gene (RPS2) derived from Arabidopsis thaliana The DNA or a fragment thereof is introduced into higher plant cells and expressed according to a known method.

As a result, the plant can be imparted with stronger and stronger resistance as compared with the case where the glucan elicitor receptor is introduced and expressed.

In addition, it has been proposed that fungi that can infect plants generally have a sublesser, and that plants have acquired the ability to suppress the resistance of plants to fungi. However, even in these cases, the DNA encoding the chimeric receptor of the present invention or a fragment thereof is introduced and expressed so that the chimeric receptor functions, or these DNAs are modified or the expression level is reduced. By making adjustments, it is possible to create new plants that are more resistant and resistant to power.

Furthermore, the DNA of the present invention or a fragment thereof is introduced into a higher plant together with a gene or a trait that enhances resistance to fungi such as dalkanase that exhibits mold resistance, and expressed in higher plants. Strong mold resistance can be imparted to plants.

The DNA sequence encoding dalcanase may be a protein containing an amino acid sequence represented by SEQ ID NO: 11 or 13, or at least one amino acid in the amino acid sequence is deleted, substituted, or added. Alternatively, there may be mentioned a DNA containing an inserted amino acid sequence and a nucleotide sequence encoding a protein having dalkanase activity. The DNA contains all degenerate isomers. An example of such a degenerate isomer is a DNA containing the nucleotide sequence shown in SEQ ID NO: 12 or 14.

As a method for introducing a gene into a plant cell or a plant, generally known methods, for example, Eds., Blackwel 1 Scientific Publications, 1988, "Plant genetic transformation and gene expression; a laboratory manual".

Examples of the gene transfer method include a biological method using a virus, an agrobacterium method, an electroporation method, a physical and chemical method, and a polyethylene glycol method. Method, microinjection method, particle gun method, dextran method and the like.

In general, when the plant into which DNA is to be introduced is a dicotyledon, a biological method using an agrobacterium is preferable.For a dicotyledon that is hardly infected with a monocotyledon, agrobacterium, A physical-chemical method such as an electroporation method is preferred. As the plant material for DNA introduction, it is possible to select appropriate materials from leaves, stems, roots, petals, tubers, protoplasts, calli, pollen, seed embryos, shoot primordia, etc., according to the method of introduction, etc. I can do it.

In general, when DNA is introduced into cultured plant cells, protoplasts are used as a material, and the DNA is introduced by a physical or chemical method such as an electo-poration method or a polyethylene glycol method. In contrast, when introducing DNA into plant tissue, leaves, stems, roots, petals, tubers, protoplasts, calli, pollen, seed germs, shoot primordia, etc., and preferably leaves and stems are used as materials. DNA can be introduced by a biological method using a viral protein, a physical or chemical method such as a particle gun method, or a microinjection method, or preferably by a biological method using an agrobacterium. Will be

Furthermore, in order to regenerate a plant from a plant tissue or plant cell into which a DNA sequence encoding a chimeric receptor has been introduced, such a plant tissue or plant cell can be regenerated using a medium such as an MS medium containing an appropriate plant growth regulator. It is sufficient to culture in. The rooted seedlings can be transformed into plants by transplanting them to soil and cultivating them.

By introducing and expressing the DNA sequence encoding the chimeric receptor by the method described above, it is possible to confer resistance to pathogenic fungi or to enhance resistance to pathogenic fungi. I can do it. Examples of such a plant include a plant whose cell wall is infected with a pathogenic mold containing glucan. Specifically, solanaceous plants, Examples include leguminous plants, asteraceae plants, dianthus plants, and gramineous plants, and more specifically, tobacco, soybean, potato, rice, chrysanthemum, carnation, and the like. Never.

As the pathogenic fungi, those containing glucan in the cell wall component are preferable.Specifically, Phytophthora, Rhizoctonia, Pyricularia, Puccinia, Fusarium, Uromyces, Botrytis, Alternaria, etc. ¾: Can be mentioned, and more specifically, Phytophthora nicotianae, Rhizoctonia sol am, Pyricularia oryzae, Puccinia horiana, Fusarium oxysporum, Fusarium roseum, Fusarium tricinctum, Uromyces dianthi, Botrytis cinerea, Alternaria dianthi, etc. But it is not limited to these.

According to the present invention, a DNA sequence encoding a chimeric receptor introduced into a plant can be inherited from a plant to a plant and to a progeny via a seed. Therefore, even in seeds formed from pollen or ovary of the plant of the present invention, the introduced DNA sequence is present, and the genetic trait can be passed on to the progeny. Therefore, a plant into which the DNA encoding the chimeric receptor of the present invention has been introduced can grow without losing resistance to pathogenic fungi by growing with seeds. In addition, mass growth methods using plant tissue culture methods and conventional methods such as tree cutting, tree cutting, grafting, and stock splitting do not result in loss of resistance to pathogenic fungi, and therefore, growth is not a problem. It is possible. Whether or not the transformed plant has resistance to fungi will be examined based on the presence or absence of a plant resistant response to glucan elicitor and the test for inoculation of vinegar. In the resistance reaction test by adding elicitor, add the dalcan elicitor solution to the surface of the leaf, or infill the glucan elicitor solution with the syringe from the back of the leaf, and observe the browning reaction that appears on the leaf surface after a certain period of time. It is performed by examining the accumulation of fluorescent substances (phytoarexins) that are excited by UV light. In addition, in the mold inoculation test, resistance can be examined by the following inoculation methods. Inoculation methods are broadly classified into two types: inoculation methods for above-ground diseases and inoculation methods for underground diseases.

I. The following methods can be used to inoculate the above-ground disease. (1) Spray inoculation method: Spray a plant with a suspension of pathogenic bacteria or spores and incubate it in an inoculation box at an appropriate temperature and saturation for a certain period of time to check the extent of the disease.

(2) Spray inoculation method: The spores collected from the lesions can be knocked down onto the plant with a brush or sprayed with a small duster. After inoculating the inoculated plants for 24 hours in an inoculation box adjusted to the appropriate temperature for infection, perform the same procedure as spray inoculation.

(3) Wound inoculation method: Inoculate a spore suspension or a piece of cultured bacterial flora into the cut end of a branch or the area where the bark has been punched out with a cork boiler.

(4) Needle inoculation method: Damage the plant with a needle or needle bundle soaked in a suspension of pathogenic bacteria and inoculate it.

(5) Cell adhesion inoculation method: For pathogenic bacteria that do not form spores on the medium, adhere the cultured mycelial mass or sclerotium to the surface of the plant and inoculate it.

(6) Damaged plant inoculation method: Instead of inoculating pathogens directly, place the damaged plant on the plant or hang the damaged leaf stem on the plant, and use the mycelium or falling spores that grow from there. Inoculate.

I I. The following methods can be used as inoculation methods for underground diseases.

(1) Soil irrigation inoculation method: Spores or crushed mycelium suspension are irrigated into the soil and inoculated.

(2) Soil admixture inoculation method: Inoculate the soil with filamentous fungi cultured on bran, grain, rice straw, wheat straw, etc.

(3) Buried inoculation of diseased plants: The residue of diseased plants is buried in soil to create soil contaminated with pathogenic bacteria. At the stage where the diseased plant residue has rotted, mix the soil well and sow or transplant the seedlings.

(4) Contaminated soil inoculation method: If there is a frequent site of a specific soil disease, collect the soil, mix well, put it in a pot or vat, sow it there, or transplant a seedling.

(5) Root inoculation method: Prepare a suspension of bacterial or filamentous fungal spores, immerse the roots of healthy seedlings in this for 3 to 10 minutes, and then transplant them to pots.

A resistance test can be performed by the above inoculation method. More specifically, the disease-causing fungus can be detected by directly inoculating mycelium into a plant individual and increasing the number of lesions. In addition, the resistance can be tested by inoculating zoospores of the disease-causing fungus and changing the lesions. In addition, plague bacteria For other types of soil bacteria, the cultured cells are introduced into the soil, seeded or planted in the soil, and the resistance to viability can be tested by observing the phenomenon of withering. However, the present invention is not limited to this. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a schematic diagram of the construction of the chimeric DNA of the present invention.

FIG. 2 is a schematic diagram of the construction of plasmid pBI (Hm) -ER.

FIG. 3 is a schematic diagram of the construction of plasmid pBI (Hm) -LER.

FIG. 4 is a schematic diagram of a plasmid containing DNA encoding about 1/3 of the N-terminal side of RPS2.

FIG. 5 is a schematic diagram of the construction of plasmid pBI (Hm) -CER.

FIG. 6 is a schematic diagram of the construction of plasmid pBI (Hm) -IER.

FIG. 7 is a schematic diagram of the construction of plasmid pBI (Hm) -ISER. BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in more detail with reference to Examples, but the technical scope of the present invention is not limited by these Examples.

In the following examples, the dalcan elicitor receptor will be abbreviated as “ER”. The meaning of the abbreviations in FIGS. 2 to 7 is as follows.

Restriction enzyme cleavage site:

B: BamHI, Xb: Xbal, SI: Sail, RI: EcoRI, RV: EcoRV, Xh: Xhol, Sph: Sphl, Sp: Spel, Scl: Sac I, Sra: Smal, Nsp: Nspl, N: Notl, Nsi : Nsil, -Nc: Ncol, H3: Hindlll

(Sin): DNA was inserted into the Smal site, and the Smal recognition sequence was crushed.

dScI, dB: Sac I and BamHI sites are crushed, respectively.

L: leader sequence of 3-glucanase

pNOS: N0S promoter, p35S: Cauliflower mosaic virus 35S promoter

Nos-T: N0S NPTII: Kanamycin resistance gene, Hm: Hygromycin resistance gene RB, LB: Ti-plasmid right border DNA sequence, Ti-plasmid left border DNA sequence, respectively

[Example 1] Construction of vector-plasmid

Various vectors were prepared by the following procedure. Reagents used for preparing plasmids such as restriction enzymes and linkers were those manufactured by Takara Shuzo unless otherwise specified. DH5a (manufactured by BRL) was used for amplification and selection of the vector in E. coli.

(1) Preparation of pBI (Hm) -ER (Figure 2)

Cleavage of the BamHI and Sail sites in the multilinker of plasmid pER23-1 [N. Umemoto et al. (1997) Pro Natl. Acad. Sci. USA 279: 141] with restriction enzymes BamHI and Sail. A fragment containing the full-length ER gene flanked by both restriction enzyme sites was obtained. This fragment was separated from agarose gel, the plant-expressed binary vector plasmid pBI121 (Clontech) was cut with BamHI and Sacl, and the following was used using an automatic nucleic acid synthesizer (Applied Biosystems). The synthetic phospholipid DNA (SEQ ID NOS: 15 and 16) shown in Table 1 was synthesized.

The obtained linker DNA was cloned into BamHI and Sail sites of the pBI linker prepared by annealing and ligation to obtain pBI-ER.

5'-CTAGAGGATCCGGTACCCCCGGGGTCGACGAGCT-3 '(SEQ ID NO: 15)

5'-CGTCGACCCCGGGGGTACCGGATCCT-3 '(SEQ ID NO: 16)

Furthermore, a fragment containing the full-length ER gene obtained by digesting pBI-ER with BamHI and Sacl was separated from agarose gel, and plant-expressed binary-one-vector-plasmid pIG12-Hm [Nakamura et al. (1990, Plant Biotechnology II ( 20), pp.123-132, obtained from Prof. Kenzo Nakamura of Nagoya University.] And ligated with plasmid fragments cut with BamHI and Sacl to obtain the desired vector pBI (I½) -ER. I got

As a result of determining the base sequence from the obtained vector, the one represented by SEQ ID NO: 2 was obtained. The amino acid sequence recoded by the nucleotide sequence represented by SEQ ID NO: 2 is shown in SEQ ID NO: 1.

Escherichia coli DH5a EKB633, into which plasmid pER23-1 has been introduced, is Deposited at the Institute of Biotechnology, Industrial Technology (1-3 1-3, Higashi, Tsukuba, Ibaraki, Japan) on June 15, 1994, and its microorganism accession number is FERM BP-4699.

(2) Preparation of pBI (Hm) -LER (Fig. 3)

Hindlll sites at two sites in PER23-1 were cut with Hindlll and self-ligated to delete the Hindlll site and beyond in the ER. Next, the remaining two Notl sites were cut with Notl, filtered in with Klenow fragment, and the BamHI linker was introduced to create a new BamHI site. From this, a plasmid fragment containing the part after Hindlll of the ER gene obtained by cutting pER23-1 with BamHI and Hindlll was ligated to the fragment containing the part before Hindlll of the ER gene cut out with BamHI and Hindi11.

Further, DNA for synthetic linker (SEQ ID NOs: 17 and 18) derived from the untranslated 5 'end of soybean 3-glucanase shown below, which was cleaved with Xbal and BamHI and synthesized using an automatic nucleic acid synthesizer, was One ring and ligation were performed to obtain pLER.

5'- CTAGACTTCTTTCCTCAACCTTCTTTCTTCTTATATATTCGAAC-3 '(SEQ ID NO: 17) 5'-GATCGTTCGAATATATAAGAAGAAAGAAGGTTGAGGAAAGAAGT-3' (SEQ ID NO: 18) did. Furthermore, a fragment containing the ER gene containing the leader sequence obtained by cutting with Xbal and Sacl was separated from the agarose gel, and a plasmid fragment obtained by cutting the plant-expressed binary-vector-plasmid PIG121-Hm with Xbal and Sacl And the desired vector pBI (Hm) -LER was obtained.

As a result of determining the base sequence from the obtained vector, the one represented by SEQ ID NO: 4 was obtained. The amino acid sequence encoded by the nucleotide sequence represented by SEQ ID NO: 4 is shown in SEQ ID NO: 3.

(3) Acquisition of cDNA clone encoding about 1/3 of RPS2N end from Arabidopsis thai iana

Arabidopsis thai iana cDNA was obtained by spreading 200,000 plaques of Clontech cDNA library on E. coli C600hfl and spreading them on 10 cm diameter LB plates. The phage was obtained by collecting the phage from the cells. Using the phage collected from the two plates as one pool, five pools of phage library were obtained. Phage DNA was recovered from each pool according to Ishida's method (Gene Expression Experiment Manual, Ishida and Ando, Kodansha Scientific).

For the N-terminal cDNA of PRPS2, PCR (using Takara Shuzo's EX-Taq kit) was performed using about 1 microgram of phage DNA (pool # 1 to # 5) as a template using the following PCR primer set. ) And cloned into pBluescript SKI I (+) plasmid.

(0 sense primer: 5'-TGGCATGCGATGGATTTCATCTCATCTCTT-3 '(SEQ ID NO: 19)

Antisense primer: 5'-GGGAATTCACTCCGCGAGCCGGCGAAT-3 '(SEQ ID NO:

20)

(ii) Sense primer: 5'-ACAAGTAAAAGAAAGAGCGAGAAATCATCGAAATG-3 '(SEQ ID NO: 21)

Antisense primer: 5'-AGCCATGGCTCCTCCTAAAGTGAT-3 '(SEQ ID NO:

twenty two)

PCR was performed with the primer set (i) or (ii).

The amplified DNA was inserted into the Smal site of pBluescriptSKII (+) after phosphorylation, and plasmid DNAs (pRPS2-6, pRPS2-14 and pRPS2-16) shown in FIG. 4 were obtained. These were confirmed by DNA sequencing to be identical to the sequence reported [M. Mindrinos et al. (1994) Cell 78: 1089] except for a portion of the primer.

(4) Preparation of pBI (Hm) -CER (Figure 5)

PRPS2-14 was digested with the restriction enzyme Xbal, and the Xbal fragment was removed by self-ligation. A BaraHI linker was inserted into the EcoRV site of this plasmid, and a Sacl linker was inserted into the Xhol site. This plasmid was cut with BamHI and Sal1, and the entire ER cut out from pER23-l with BamHI and Sail was cloned. From this plasmid, a chimeric DNA containing a part of RPS2 and the entire ER was purified. Ensure that the junction between RPS2 and ER matches the translation frame by DNA sequence. And confirmed.

On the other hand, pSpellinker is inserted into the EcoRV site of pRPS2-6. Next, the purified DNA fragment was cloned into the Xbal and Sad cleavage sites of pRPS2-6. The plasmid containing the chimeric gene fused to the 13 'of the 5' end of RPS-2 and the full length of the ER is cut with Spel and Sad, and the chimeric gene is cloned into pIG12 Hm plasmid, and the desired vector pBI ( Hm) -CER was obtained.

As a result of determining the nucleotide sequence from the obtained vector, the one represented by SEQ ID NO: 6 was obtained. The amino acid sequence encoded by the nucleotide sequence represented by SEQ ID NO: 6 is shown in SEQ ID NO: 5.

(5) Preparation of pBI (Hm) -IER (Figure 6)

From PER23-1 plasmid, ER cDNA encoding 537 amino acids at the N-terminal is purified by BamHI + NspI. On the other hand, RPS2 cDNA is purified using pRPS2-16 EcoRV + Sphl. These two DNA fragments were cloned into the BamHI and Smal sites of pBluescriptSKI I (+) plasmid. This plasmid is called pERRPS-8. It was confirmed that the translation frame was joined by the DNA sequence at the junction between RPS2 and ER. The fragment containing the fusion gene of the part encoding the elicitor-binding region and the part encoding the RPS2 partial length obtained by digesting pERRPS-8 with Spel and Sacl was separated from agarose gel and separated from the plant-expressed binary. The plasmid pIG121-Hm was digested with Xbal and Sacl and ligated with the plasmid fragment to obtain the desired vector pBI (Hm) -IER.

The nucleotide sequence was determined from the obtained vector, and as a result, the one represented by SEQ ID NO: 8 was obtained. The amino acid sequence encoded by the nucleotide sequence represented by SEQ ID NO: 8 is shown in SEQ ID NO: 7.

(6) Preparation of pBI (Hm) -ISER (Figure 7)

The pERRPS-8 plasmid was cut with Nsil + Notl, and the Notl-Nsil adapter shown below was inserted. The DNA sequence confirmed that the translation frames of ATG and ER in the adapter matched. Cut this plasmid with EcoRV + Sall, Purify the RPS2 / ER chimeric DNA.

5'-GGCCGCGATATCATGAATGCA-3 '(SEQ ID NO: 23)

5'-TTCATGATATCGC-3 '(SEQ ID NO: 24) A synthetic DNA derived from the untranslated 5' end of 1-3-glucanase shown below was prepared and inserted into the Notl and EcoRI sites of pBluescriptSKII (+). Plasmid pL was prepared.

5'-GGCCGCCTTCTTTCCTCAACCTTCTTTCTTCTTATATATTCGAAC-3 '(SEQ ID NO: 25) 5'-AATTGTTCGAATATATAAGAAGAAAGAAGGTTGAGGAAAGAAGGC-3' (SEQ ID NO: 26) Plasmid Entered. The plasmid prepared here is called pSERRPS-8. A fragment containing the fusion gene for the elicitor-binding region obtained by cutting pSERRPS-8 with Spel and Sacl and the portion encoding the RPS2 partial length was separated from agarose gel and separated from a plant-expressed binary vector plasmid. The plasmid pIGI2Hra was ligated with a plasmid fragment cut with Xbal and Sacl to obtain the desired vector pBI (Hm) -ISER.

As a result of determining the base sequence from one of the obtained vectors, one represented by SEQ ID NO: 10 was obtained. The amino acid sequence encoded by the nucleotide sequence represented by SEQ ID NO: 10 is shown in SEQ ID NO: 9.

[Example 2] Introduction of chimeric receptor gene into Arabidopsis thaliana plant (1) Preparation of Agrobacterium infectious solution

Transformation vector containing the re-chimeric receptor gene [pBI (Hm) -CER, pBI (Hm) -ISER] and the control vector containing the ER gene as a control [pBI (Hm) -ER] -introduced Agrobacterium tumefaciens EHAIOI was inoculated into 3 ml of YEB medium, cultured at 28 ° C for 16 hours, collected by centrifugation, suspended in 10 ml of infection medium, and used for transformation. Agrobacterium infection solution. The medium compositions of the YEB medium and the infection medium are described below.

YEB medium: 5g / l beef extract, lg / 1 yeast extract, 5g peptone, 5g / l skull —S, 2mM magnesium sulfate

Infection medium; 1/2 concentration of MS [Murashige & Skoog (1962) Physiol. Plant. 15: 473] Inorganic salts and vitamins of the medium, 15 g / l sucrose, lOg / 1 glucose, lOmM MES (pH5.4)

(2) Method for producing Arabidopsis transformed plant

The seeds of Arabidopsis thai iana L. strains, Ransburg and Colombia, were aseptically sowed on MSHF medium and cultured for 3 weeks. The root extended to 2-3 cm was cut into about 5 thighs, transplanted to a callus induction medium, and cultured for 2 days. The root explant was immersed in an Agrobacterium-infected solution for 10 minutes, and excess infection solution adhering to the root explant was wiped off on a filter paper, and then transplanted to a culture medium containing arabid and cultured for 2 days in the dark. The root pieces were transplanted to an arabid regeneration medium and cultured for 2 days, and then transplanted to an arabid selection regeneration medium and cultured for 1 week. Thereafter, the cells were transplanted to a fresh regeneration medium for selection of arabido every week, and the culture was continued until a regenerated seedling was obtained.

Seedlings whose shoots and leaves reached about 1 cm were transplanted to arabid rooting medium and continued to be cultured. The presence of the transgene in the plant whose roots extended to 1-2 cm was confirmed by PCR, and the transformant was used. The culture was performed under the conditions of 22t: 16 hours lighting and 8 hours no lighting unless otherwise specified. The components of the medium used for the culture are described below.

MSHF medium: MS [Murashige & Skoog (1962) Physiol. Plant. 15: 473] Inorganic salts and vitamins of the medium, 20 g / l sucrose, 2 g / l gellan gum, 5 mM MES (pH 6.2)

Callus induction medium: Inorganic salts and vitamins of MS medium, 20 g / l sucrose, 0.5 mg / 12,4-dichlorofluorophenoxyacetic acid, 0.1 mg power ricetin, 2 g / l gellan gum, 5 mM MES (pH 6.2)

Arabide co-culture medium: Inorganic salts and vitamins of MS medium, 20 g / l sucrose, 0.5 mg / 12,4-dichlorophenoxyacetic acid, 0.1 mg / l strength ricetin, 0.2 mM acetosyringone, 2 g / l gellan gum, 5mM MES (pH 6.2)

Arabido regeneration medium: Inorganic salts and vitamins of MS medium, 20 g / l sucrose, lmg / 1 2-isopentenylamine, 0.15 mg / 1 indole-250 mg / 1 Cefotaxime, 2g / l gellan gum, 5mM MES (pH 6.2)

Regeneration medium for selection of arabide: Inorganic salts and vitamins of MS medium, 20 g sucrose, lmg / 1 2-isopentenylamine, 0.15 mg / l indoleacetic acid, 2 g / l gellan gum, 50 mg / 1 kanamycin, 250 mg / l Cefotaxime , 5mM MES (pH 6.2)

Arabidopsis root medium: Inorganic salts and vitamins of MS medium, 20 g / l sucrose, 2 mg / l naphthalene acetic acid, 2 g / l gellan gum, 250 mg / l Cefotaxime, 5 mM MES (pH 6.2)

[Example 3] Introduction of chimeric receptor gene into tobacco plant

(1) Preparation of Agrobacterium infection solution

A vector for transformation containing the chimeric receptor gene [pBI (Hni) -IER] and a vector for transformation containing the ER gene [pBI (Hm) -LER] were used as controls by electroporation. The introduced Agrobacterium tumefaciens EHA101 was inoculated into 3 ml of YEB medium, cultured at 28 ° C for 16 hours, collected by centrifugation, suspended in 10 ml of infection medium, and transformed into an agrobacterium for transformation. This was a terium infection solution. The medium compositions of the YEB medium and the infection medium are described below.

YEB medium: 5g / l beef extract, lg / 1 yeast extract, 5g / l peptone, 5g / l sucrose, 2mM magnesium sulfate

Infection medium: 1/2 concentration of MS [Murashige & Skoog (1962) Physiol. Plant. 15: 473] Inorganic salts and vitamins of the medium, 15 g / l sucrose, lOg / 1 glucose, 10 mM MES (pH 5.4)

(2) Method for producing transgenic tobacco plant

Leaves of the samsun line of tobacco (Nicotiana tabacum L.) grown in a greenhouse were sterilized and cut into approximately 1 cm squares. The leaf cuttings were floated in an Agrobacterium infection solution for 10 minutes, and excess infection solution attached to the leaf pieces was wiped off on a filter paper, then transplanted to a tobacco co-culture medium, and cultured for 2 days in the dark. The leaf pieces were transplanted to a tobacco regeneration medium and cultured for 1 week, and then transplanted to a regeneration medium for tobacco selection and cultured for 2 weeks. After that, transfer to fresh regeneration medium for tobacco selection every two weeks and continue culturing until regenerated seedlings are obtained. I did.

Seedlings whose stems and leaves became about lcm were transplanted to tobacco rooting medium and cultured. The presence of the transgene in the plant whose roots extended to l-2 cm was confirmed by PCR and used as a transformant. The culture was performed under the conditions of 25 ° C, 16 hours of illumination and 8 hours of no illumination unless otherwise specified. The components of the medium used for the culture are described below.

Tobacco co-culture medium: MS [Murashige & Skoog (1962) Physiol. Plant. 15: 473] Inorganic salts and vitamins of the medium, 30 g sucrose, O.lmg / 1 naphthalene acetic acid, lmg benzyladenine, 0.2 mM acetocillingone, 8 g / l Agar, 5mM MES (pH5.8)

Tobacco regeneration medium: MS medium inorganic salts and vitamins, 30 g / l sucrose, 0.1 mg / 1 naphthalene acetic acid, lmg / 1 benzyl adenine, 250 mg cefotaxime, 8 g / l agar, 5 mM MES (pH 5.8)

Regeneration medium for tobacco selection: MS medium inorganic salts and vitamins, 30 g / l sucrose, 0.1 mg / 1 naphthalene acetic acid, lmg / 1 benzyladenine, 100 mg / 1 kanamycin, 250 mg cefotaxime, 8 g agar, 5raM MES (pH5 .8)

Tobacco rooting medium: Inorganic salts and vitamins in MS medium, 30g / l sucrose, 250rag / l Cefotaxime, 8g agar, 5mM MES (pH5.8)

[Example 4] Hypersensitive reaction test of transformed Arabidopsis thaliana

(1) Hypersensitivity test of transgenic Arabidopsis (strain Landsberg)

Among the prepared chimeric receptor genes, an Arabidopsis transformant (CER) in which high expression of the gene containing the nucleotide sequence represented by SEQ ID NO: 6 was confirmed, and an Arabidopsis trait in which high expression of ER protein 'was confirmed as a control The induction of a hypersensitive response by soybean elicitor was examined for the leaves of the transformant (ER; SEQ ID NO: 2) and untransformed Arabidopsis thaliana (control).

10 mM MgCl ₂ fungal cells was dissolved in a solution wall from Dar cans elicitor (lOO g / ml), and 10 mM MgCl ₂ solution, infiltrated rate syringe without a back or we needle leaves of Arabidopsis plants grown in a greenhouse, The cells were cultured at 22 ° C for 7 days in a cycle of 16 hours light and 8 hours dark, and changes appearing on the leaf surface were observed. Mold cells used here The ηππππηπηππηπ glucan elicitor derived from the wall was obtained according to the method of Umeki et al. [N. Umemoto et.

Natl. Acad. Sci. USA 94: 1029].

When glucan elicitor was infiltrated into untransformed Arabidopsis thaliana, no change was observed on the leaf surface. This indicates that Arabidopsis cannot recognize Dalkan elicitor. In Arabidopsis thaliana plants expressing the Dalcan elicitor receptor (ER), a weak browning reaction (resistance reaction) was observed on the leaf surface of 2 of the 6 clones tested, whereas in Arabidopsis thaliana plants expressing the chimeric receptor (CER), A strong browning reaction (resistance reaction) was observed in 8 out of 12 clones tested, and a weak browning reaction was observed in 1 clone (Table 1).

Table 1. Hypersensitivity test of transgenic Arabidopsis (strain Landsberg)

From mold cell wall

Glucan elicita 10mM MgCl

-Le 1

-Le 2

-Le 3

-Le 4

Lily 5

-Le 6

-Le 7

'Le 8

-Le 9

-Le 10

-Le 11

Relais 12

ER 1

ER 2

ER 3

ER 4

ER 5

ER 6+

CER 1 ++

CER 2 ++

CER 3 ++

CER 4 ++

CER 5

CER 6 +++

CER 7 +++

CER 8 ++

CER 9

CER 10

CER 11 ++

CER 12

: No change, +: Browning reaction small, ++: During browning reaction, +++: Browning reaction large These results indicate that not only the soybean-derived ER protein was expressed alone, but also a chimera composed of the ER and the N-terminal domain of the Arabidopsis thaliana RPS2 protein, which is predicted to contain a signal transduction domain. By expressing the receptor in Arabidopsis thaliana, glucan elicitor that the host plant could not recognize was recognized, and it was proved that expression of the chimeric receptor was more advantageous for recognition of glucan elicitor. This indicates that signal transduction after glucan elicitor binding to the receptor is more efficient when the chimeric receptor is expressed than when the ER protein alone is expressed.

The present invention recognizes the contact and invasion of plant pathogens such as molds having a glucan structure on the cell wall, etc., and thereby induces a resistance reaction that is deeply involved in disease resistance more strongly than when only the ER protein is expressed. And a plant that can be used. By producing a plant that strongly induces the resistance response caused by such dalcan elicitor, it is possible to breed plants that exhibit strong resistance to phytopathogenic fungi such as mold.

(2) Hypersensitivity test of transgenic Arabidopsis thaliana Colombia

Among the prepared chimeric receptor genes, a transgenic Arabidopsis thaliana (ISER) in which high expression of the gene containing the nucleotide sequence represented by SEQ ID NO: 10 was confirmed, and a non-transformed Arabidopsis thaliana (control) The induction of a hypersensitive response by soybean elicitor was examined on the leaves.

LOMM MgC l ₂ fungal cells was dissolved in a solution wall from Dar cans elicitor (lOO ^ g / ral), and infill the 10 mM MgC l ₂ solution in a syringe without a back or we needle leaves of Arabidopsis plants grown in a greenhouse The cells were cultured at 22 ° C for 7 days on a 16-hour light, 8-hour dark cycle, and the changes appearing on the leaf surface were observed. The mold cell wall-derived dalcan elicitor used here was obtained according to the method of Umeki et al. [N. Umemoto et al. (1997) Pro Natl. Acad. ScI. USA 94: 1029].

When glucan elicitor was infiltrated into untransformed Arabidopsis thaliana, no change was observed on the leaf surface. This means that Arabidopsis Indicates that it cannot recognize Dalcan elicita. In Arabidopsis thaliana plants expressing chimera receptor (ISER), a strong browning reaction (resistance reaction) was observed in 3 out of 5 clones tested (Table 2).

Table 2 Hypersensitivity test of transgenic Arabidopsis thaliana

Mold cell wall-derived 10 mM MgCl ₂

Dal Power

Control 1

Control 2 —-Control 3 — One

I SER 1 +++-

I SER 2 I

I SER 3 ++-

I SER 4 +++ one

I SER 5 I

—: No change, +: Browning reaction small, ++: Browning reaction, +++: Browning reaction large From these results, it can be concluded that ER lacking about 1/3 of N-terminal and domain of signal transduction Expression of a chimeric receptor composed of the N-terminal domain of Arabidopsis thaliana RPS2 protein, which is expected to contain, in Arabidopsis thaliana proved that glucan elicitor, which cannot be recognized by the host plant, was recognized. . Further, the present invention provides a plant which can recognize contact and invasion of plant pathogens such as mold having a glucan structure on a cell wall or the like. By producing a plant that strongly induces the resistance response caused by such a glucan elicitor, it becomes possible to produce a plant that exhibits strong resistance to phytopathogenic bacteria such as power plants.

[Example 5] Hypersensitive reaction test of transgenic tobacco

Among the prepared quinula receptor genes, a tobacco transformant (IER) in which high expression of the gene containing the nucleotide sequence represented by SEQ ID NO: 8 was confirmed, and as a control, tobacco transformation in which high expression of ER protein was confirmed The induction of the hypersensitive response by soybean elicitor was examined for the leaves of the tobacco (LER; SEQ ID NO: 4) and the untransformed tobacco (control).

The leaves of tobacco plants grown in the greenhouse are cut off with a petiole, and plastic that transmits light is used. Place it in the box. Place a cut silicon tube with a diameter of 5 mm and a height of 5 mm on the front side of the leaf so that it can hold the solution. Here, mold cell wall-derived dalcan elicitor UOO g / ml dissolved in 10 mM MgCl ₂ solution, chemically synthesized dalcan elicitor-(100 mg / ml), and 10 mM MgCl ₂ solution as a control The cells were cultured in a humid condition at 25 ° C. for 7 days in a light-dark and 8-hour dark cycle to prevent drying. Chemically synthesized darcan elicita (/ 3-D-glucohexaoside) [N. Hong and T. Ogawa (1990) Tetrahedron Lett. 31: 3179] was obtained from Prof. Ogawa (RIKEN and the University of Tokyo). Using. After the cultivation, the accumulation of tobacco and huitaurexin was observed on a UV illuminator (Funakoshi) except for the silicon tube.

No change was observed when Dulcan elicita was added to untransformed tobacco. This indicates that untransformed tobacco cannot recognize Dalkan elicita. In ER-expressing tobacco plants, only a small amount of fluorescent substance (phytoalexin) was accumulated in one of the five clones tested. In IER-expressing tobacco plants, three out of five clones tested In this case, a significant amount of fluorescent substance (phytorexin) was accumulated, and a small amount of fluorescent substance (phytoalexin) was accumulated in one clone (Table 3). Table 3 Hypersensitive reaction test of transgenic tobacco

Mold cell wall-derived chemical synthesis 10 mM MgCl glucan elicita glucan elicita control 1

Control 2

LER 1

LER 2

LER 3

LER 4

LER 5

IER 1 +++ ++

IER 2 +++ +++

IER 3 +++

IER 4

IER 5

1: No change, +: Small accumulation of fluorescent substance, ++: During accumulation of fluorescent substance

+++: Large accumulation of fluorescent material These results indicate that not only the soybean-derived ER protein was expressed alone, but also that the chimera receptor composed of the ER and the N-terminal domain of the Arabidopsis-derived RPS2 protein was used in host plants other than soybean and Arabidopsis. (Tobacco), it was proved that dalcan elicitor, which the host plant (Tobacco) could not recognize, was recognized. In plants other than Arabidopsis thaliana, the chimeric receptor containing the N-terminal domain of RPS2 could be used. It was shown that signal transduction was performed more efficiently than expression of the gene encoding the ER protein. Further, the present invention provides a DNA encoding a chimeric receptor capable of recognizing contact and invasion of plant pathogens such as mold having a glucan structure in a cell wall or the like, and a plant body, whereby a firefly lexin which is deeply involved in disease resistance is provided. Resistance reactions such as induction are more intense than when the ER protein alone is expressed. By producing a plant that strongly induces the resistance response caused by such a glucan elicitor, it is possible to breed plants that exhibit strong resistance to phytopathogenic fungi such as mold. Industrial applicability

According to the present invention, there is provided a DNA encoding a chimeric protein (a chimeric receptor-1) comprising a domain containing at least an elicitor-binding site of an elicitor-receptor and a domain containing a signaling motif of a plant-derived resistance gene. And a method for producing the plant disease-resistant plant.

The chimeric receptor of the present invention can be used for elucidating the mechanism of disease resistance to mold.

The DNA containing the base sequence encoding the quinula receptor of the present invention and fragments thereof are useful as materials for establishing a technique for breeding mold-resistant plants. That is, when the DNA of the present invention or a fragment thereof is introduced into various plants and expressed, the resistance of the plants to mold can be increased. Sequence listing

SEQ ID NO: 1

Sequence length: 667

Sequence type: amino acid

Topology: linear

Sequence type: protein

Array:

Val Asn lie Gin Thr Asn Thr Ser Tyr lie Phe Pro Gin Thr Gin Ser

1 5 10 15

Thr Val Leu Pro Asp Pro Ser Lys Phe Phe Ser Ser Asn Leu Leu Ser

20 25 30

Ser Pro Leu Pro Thr Asn Ser Phe Phe Gin Asn Phe Val Leu Lys Asn

35 40 45

Gly Asp Gin Gin Glu Tyr He His Pro Tyr Leu lie Lys Ser Ser Asn

50 55 60

Ser Ser Leu Ser Leu Ser Tyr Pro Ser Arg Gin Ala Ser Ser Ala Val 65 70 75 80 lie Phe Gin Val Phe Asn Pro Asp Leu Thr lie Ser Ala Pro Gin Gly

85 90 95

Pro Lys Gin Gly Pro Pro Gly Lys His Leu lie Ser Ser Tyr Ser Asp

100 105 110

Leu Ser Val Thr Leu Asp Phe Pro Ser Ser Asn Leu Ser Phe Phe Leu

115 120 125

Val Arg Gly Ser Pro Tyr Leu Thr Val Ser Val Thr Gin Pro Thr Pro

130 135 140

Leu Ser lie Thr Thr lie His Ser lie Leu Ser Phe Ser Ser Asn Asp 145 150 155 160

Ser Asn Thr Lys Tyr Thr Phe Gin Phe Asn Asn Gly Gin Thr Trp Leu

165 170 175 Leu Tyr Ala Thr Ser Pro 11 e Lys Leu Asn His Thr Leu Ser Glu lie

180 185 190

Thr Ser Asn Ala Phe Ser Gly I le lie Arg lie Ala Leu Leu Pro Asp

195 200 205

Ser Asp Ser Lys His Glu Ala Val Leu Asp Lys Tyr Ser Ser Cys Tyr

210 215 220

Pro Val Ser Gly Lys Ala Val Phe Arg Glu Pro Phe Cys Val Glu Tyr 225 230 235 240

Asn Trp Glu Lys Lys Asp Ser Gly Asp Leu Leu Leu Leu Ala His Pro

245 250 255

Leu His Val Gin Leu Leu Arg Asn Gly Asp Asn Asp Val Lys I le Leu

260 265 270

Glu Asp Leu Lys Tyr Lys Ser lie Asp Gly Asp Leu Val Gly Val Val

275 280 285

Gly Asp Ser Trp Val Leu Lys Thr Asp Pro Leu Phe Val Thr Trp His

290 295 300

Ser He Lys Gly lie Lys Glu Glu Ser His Asp Glu lie Val Ser Ala 305 310 315 320

Leu Ser Lys Asp Val Glu Ser Leu Asp Ser Ser Ser lie Thr Thr Thr

325 330 335

Glu Ser Tyr Phe Tyr Gly Lys Leu lie Ala Arg Ala Ala Arg Leu Val

340 345 350

Leu lie Ala Glu -Glu Leu Asn Tyr Pro Asp Val lie Pro Lys Val Arg

355 360 365

Asn Phe Leu Lys Glu Thr lie Glu Pro Trp Leu Glu Gly Thr Phe Ser

370 375 380

Gly Asn Gly Phe Leu His Asp Glu Lys Trp Gly Gly lie He Thr Gin 385 390 395 400

Lys Gly Ser Thr Asp Ala Gly Gly Asp Phe Gly Phe Gly lie Tyr Asn 0V9 SC9 0C9 529

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JA 丄 B! V uio 0Jd ΛΚΙ sAq SJV JO dj 丄 BJV OJJ dsy ns ε ι Jiu

οε sz oz

nai Λ ¾IV 911 ^ IO 9] I ^ ά 丄 ^ ID ^η3 Ί ^S ! HJ ^ IS! HS! H dsy \ V 0 \ V 0 rO / 86df / XDd S908S / 86 O Lys He Arg Asn Leu Lys Gly Phe Asp Gly Gly Asn Ser Leu Thr Asn

645 650 655

Leu Leu Trp Trp lie His Ser Arg Ser Asp Glu

660 665 SEQ ID NO: 2

Array length: 2240

Sequence type: nucleic acid

Number of chains: double strand

Topology: linear

Sequence type: cDNA to mRNA

Array features

Characteristic symbol: CDS

Location: 29..2029

Array:

TCGAATTAAA CCACCTTCAG CAACAATG GTT AAC ATC CAA ACC AAT ACA TCT 52

Val Asn He Gin Thr Asn Thr Ser

1 5

TAC ATC TTC CCT CAA ACA CAA TCC ACT GTT CTT CCT GAT CCC TCC AAA 100 Tyr lie Phe Pro Gin Thr Gin Ser Thr Val Leu Pro Asp Pro Ser Lys

10 15 20

TTC TTC TCC TCA AAC CTT CTC TCA AGT CCA CTC CCC ACA AAC TCT TTC 148 Phe Phe Ser Ser Asn Leu Leu Ser Ser Pro Leu Pro Thr Asn Ser Phe

25 30 35 40

TTC CAA AAC TTT GTC CTA AAA AAT GGT GAC CAA CAA GAA TAC ATT CAT 196 Phe Gin Asn Phe Val Leu Lys Asn Gly Asp Gin Gin Glu Tyr lie His His

45 50 55

CCT TAC CTC ATC AAA TCC TCC AAC TCT TCC CTC TCT CTC TCA TAC CCT 244 Pro Tyr Leu lie Lys Ser Ser Asn Ser Ser Leu Ser Leu Ser Tyr Pro 60 65 70

TCT CGC CAA GCC AGT TCA GCT GTC ATA TTC CAA GTC TTC AAT CCT GAT 292 Ser Arg Gin Ala Ser Ser Ala Val lie Phe Gin Val Phe Asn Pro Asp

75 80 85

CTT ACC ATT TCA GCC CCA CAA GGT CCC AAA CAA GGT CCC CCT GGT AAA 340 Leu Thr He Ser Ala Pro Gin Gly Pro Lys Gin Gly Pro Pro Gly Lys

90 95 100

CAC CTT ATC TCC TCC TAC AGT GAT CTC AGT GTC ACC TTG GAT TTC CCT 388 His Leu lie Ser Ser Tyr Ser Asp Leu Ser Val Thr Leu Asp Phe Pro

105 110 115 120

TCT TCC AAT CTG AGC TTC TTC CTT GTT AGG GGA AGC CCC TAT TTG ACT 436 Ser Ser Asn Leu Ser Phe Phe Leu Val Arg Gly Ser Pro Tyr Leu Thr

125 130 135

GTG TCT GTG ACT CAA CCA ACT CCT CTT TCA ATT ACC ACC ATC CAT TCC 484 Val Ser Val Thr Gin Pro Thr Pro Leu Ser lie Thr Thr lie His Ser

140 145 150

ATT CTC TCA TTC TCT TCA AAT GAC TCC AAC ACC AAG TAC ACC TTT CAG 532 lie lie Leu Ser Phe Ser Ser Asn Asp Ser Asn Thr Lys Tyr Thr Phe Gin

155 160 165

TTC AAC AAT GGT CAA ACA TGG CTT CTT TAT GCT ACC TCC CCC ATC AAG 580 Phe Asn Asn Gly Gin Thr Trp Leu Leu Tyr Ala Thr Ser Pro lie Lys

170 175 180

TTG AAC CAC ACC CTT TCT GAG ATA ACT TCT AAT GCA TTT TCT GGC ATA 628 Leu Asn His Thr Leu Ser Glulie Thr Ser Asn Ala Phe Ser Gly He

185 190 195 200

ATC CGG ATA GCT TTG TTG CCG GAT TCG GAT TCG AAA CAC GAG GCT GTT 676 lie Arg lie Ala Leu Leu Pro Asp Ser Asp Ser Lys His Glu Ala Val

205 210 215

CTT GAC AAG TAT AGT TCT TGT TAC CCC GTG TCA GGT AAA GCT GTG TTC 724 Leu Asp Lys Tyr Ser Ser Cys Tyr Pro Val Ser Gly Lys Ala Val Phe

220 225 230

AGA GAA CCT TTC TGT GTG GAA TAT AAC TGG GAG AAG AAA GAT TCA GGG 772 Arg Glu Pro Phe Cys Val Glu Tyr Asn Trp Glu Lys Lys Asp Ser Gly

235 240 245

GAT TTG CTA CTC TTG GCT CAC CCT CTC CAT GTT CAG CTT CTT CGT AAT 820 Asp Leu Leu Leu Leu Ala His Pro Leu His Val Gin Leu Leu Arg Asn

250 255 260

GGA GAC AAT GAT GTC AAA ATT CTT GAA GAT TTA AAG TAT AAA AGC ATT 868 Gly Asp Asn Asp Val Lys lie Leu Glu Asp Leu Lys Tyr Lys Ser lie

265 270 275 280

GAT GGG GAT CTT GTT GGT GTT GTC GGG GAT TCA TGG GTT TTG AAA ACA 916 Asp Gly Asp Leu Val Gly Val Val Gly Asp Ser Trp Val Leu Lys Thr

285 290 295

GAT CCT TTG TTT GTA ACA TGG CAT TCA ATC AAG GGA ATC AAA GAA GAA 964 Asp Pro Leu Phe Val Thr Trp His Ser I le Lys Gly lie Lys Glu Glu

300 305 310

TCC CAT GAT GAG ATT GTC TCA GCC CTT TCT AAA GAT GTT GAG AGC CTA 1012 Ser His Asp Glulie Val Ser Ala Leu Ser Lys Asp Val Glu Ser Leu

315 320 325

GAT TCA TCA TCA ATA ACT ACA ACA GAG TCA TAT TTT TAT GGG AAG TTG 1060 Asp Ser Ser Serlie Thr Thr Thr Glu Ser Tyr Phe Tyr Gly Lys Leu

330 335 340

ATT GCA AGG GCT GCA AGG TTG GTA TTG ATT GCT GAG GAG TTG AAC TAC 1108 He Ala Arg Ala Ala Arg Leu Val Leu lie Ala Glu Glu Leu Asn Tyr 345 350 355 360

CCT GAT GTG ATT CCA AAG GTT AGG AAT TTT TTG AAA GAA ACC ATT GAG 1156 Pro Asp Val lie Pro Lys Val Arg Asn Phe Leu Lys Glu Thr lie Glu

365 370 375 CCA TGG TTG GAG GGA ACT TTT AGT GGG AAT GGA TTC CTA CAT GAT GAA 1204 Pro Trp Leu Glu Gly Thr Phe Ser Gly Asn Gly Phe Leu His Asp Glu

380 385 390

AAA TGG GGT GGC ATT ATT ACC CAA AAG GGG TCC ACT GAT GCT GGT GGT 1252 Lys Trp Gly Gly He lie Thr Gin Lys Gly Ser Thr Asp Ala Gly Gly

395 400 405

GAT TTT GGA TTT GGA ATT TAC AAT GAT CAC CAC TAT CAT TTG GGG TAC 1300 Asp Phe Gly Phe Gly lie Tyr Asn Asp His His Tyr His Leu Gly Tyr

410 415 420

TTC ATT TAT GGA ATT GCG GTG CTC ACT AAG CTT GAT CCA GCA TGG GGT 1348 Phe lie Tyr Gly He Ala Val Leu Thr Lys Leu Asp Pro Ala Trp Gly

425 430 435 440

AGG AAG TAC AAG CCT CAA GCC TAT TCA ATA GTG CAA GAC TTC TTG AAC 1396 Arg Lys Tyr Lys Pro Gin Ala Tyr Ser lie Val Gin Asp Phe Leu Asn

445 450 455

TTG GAC ACA AAA TTA AAC TCC AAT TAC ACA CGT TTG AGG TGT TTT GAC 1444 Leu Asp Thr Lys Leu Asn Ser Asn Tyr Thr Arg Leu Arg Cys Phe Asp

460 465 470

CCT TAT GTG CTT CAC TCT TGG GCT GGA GGG TTA ACT GAG TTC ACA GAT 1492 Pro Tyr Val Leu His Ser Trp Ala Gly Gly Leu Thr Glu Phe Thr Asp

475 480 485

GGA AGG AAT CAA GAG AGC ACA AGT GAG GCT GTG AGT GCA TAT TAT TCT 1540 Gly Arg Asn Gln'Glu Ser Thr Ser Glu Ala Val Ser Ala Tyr Tyr Ser

490 495 500

GCT GCT TTG ATG GGA TTA GCA TAT GGT GAT GCA CCT CTT GTT GCA CTT 1588 Ala Ala Leu Met Gly Leu Ala Tyr Gly Asp Ala Pro Leu Val Ala Leu

505 510 515 520

GGA TCA ACA CTC ACA GCA TTG GAA ATT GAA GGG ACT AAA ATG TGG TGG 1636 Gly Ser Thr Leu Thr Ala Leu Glulie Glu Gly Thr Lys Met Trp Trp 525 530 535

CAT GTG AAA GAG GGA GGT ACT TTG TAT GAG AAA GAG TTT ACA CAA GAG 1684 His Val Lys Glu Gly Gly Thr Leu Tyr Glu Lys Glu Phe Thr Gin Glu

540 545 550

AAT AGG GTG ATG GGT GTT CTA TGG TCT AAC AAG AGG GAC ACT GGA CTT 1732 Asn Arg Val Met Gly Val Leu Trp Ser Asn Lys Arg Asp Thr Gly Leu

555 560 565

TGG TTT GCT CCT GCT GAG TGG AAA GAG TGT AGG CTT GGC ATT CAG CTC 1780 Trp Phe Ala Pro Ala Glu Trp Lys Glu Cys Arg Leu Glylie Gin Leu

570 575 580

TTA CCA TTG GCT CCT ATT TCT GAA GCC ATT TTC TCC AAT GTT GAC TTT 1828 Leu Pro Leu Ala Pro lie Ser Glu Ala lie Phe Ser Asn Val Asp Phe

585 590 595 600

GTA AAG GAG CTT GTG GAG TGG ACT TTG CCT GCT TTG GAT AGG GAG GGT 1876 Val Lys Glu Leu Val Glu Trp Thr Leu Pro Ala Leu Asp Arg Glu Gly

605 610 615

GGT GTT GGT GAA GGA TGG AAG GGG TTT GTG TAT GCC CTT GAA GGG GTT 1924 Gly Val Gly Glu Gly Trp Lys Gly Phe Val Tyr Ala Leu Glu Gly Val

620 625 630

TAT GAC AAT GAA AGT GCA CTG CAG AAG ATA AGA AAC CTG AAA GGT TTT 1972 Tyr Asp Asn Glu Ser Ala Leu Gin Lys lie Arg Asn Leu Lys Gly Phe

635 640 645

GAT GGT GGA AAC'TCT TTG ACC AAT CTC TTG TGG TGG ATT CAT AGC AGA 2020 Asp Gly Gly Asn Ser Leu Thr Asn Leu Leu Trp Trp lie His Ser Arg

650 655 660

AGT GAT GAA TAGAATGGAT TTGGTTATTG AACATGACAA CACTGCTGAT 2069 Ser Asp Glu

665

TTGGTTATTA TTATTATTGC TACTATTACT ATGCCAATAA AATCATCAAA TATGTCTTAC 2129 091 991 OS I 9 ^ 1 dsv usv J9S J9S sy J as "91811 S! H

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36 06 98

ID UIO ojj BIV JSS ³ Π J¾ dsy 0Jd usy Ι ^ Λ ^ io aqj en 08 L OA S9

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09 99 OS usv J9S JaS sAq an na JA 丄 OJJ S! H 911 ^ JL ^ i uio dsy \ o se

usy sA na eight aj usy aqj sj jag usy Jm OJ j naq OJJ

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OnZ V OIOIDIVDVV 1V13IVD1VD IV1VV3VV3V IIDVVVIVVI VIOVVVVDVV

621Z DVVDV1V11D 1YIVD0111V DIVVDIOVVI VVOOOIVIIV 丄丄 V 丄丄 VVVVV VVIVVIVDIV lLZ0 / S6dT / lDd S908S / 86 OM Ser Asn Thr Lys Tyr Thr Phe Gin Phe Asn Asn Gly Gin Thr Trp Leu

165 170 175

Leu Tyr Ala Thr Ser Pro lie Lys Leu Asn His Thr Leu Ser Glu lie

180 185 190

Thr Ser Asn Ala Phe Ser Gly I le I le Arg lie Ala Leu Leu Pro Asp

195 200 205

Ser Asp Ser Lys His Glu Ala Val Leu Asp Lys Tyr Ser Ser Cys Tyr

210 215 220

Pro Val Ser Gly Lys Ala Val Phe Arg Glu Pro Phe Cys Val Glu Tyr 225 230 235 240

Asn Trp Glu Lys Lys Asp Ser Gly Asp Leu Leu Leu Leu Ala His Pro

245 250 255

Leu His Val Gin Leu Leu Arg Asn Gly Asp Asn Asp Val Lys I le Leu

260 265 270

Glu Asp Leu Lys Tyr Lys Ser lie Asp Gly Asp Leu Val Gly Val Val

275 280 285

Gly Asp Ser Trp Val Leu Lys Thr Asp Pro Leu Phe Val Thr Trp His

290 295 300

Ser lie Lys Gly lie Lys Glu Glu Ser His Asp Glu He Val Ser Ala 305 310 315 320

Leu Ser Lys Asp Val Glu Ser Leu Asp Ser Ser Ser lie Thr Thr Thr

325 330 335

Glu Ser Tyr Phe 'Tyr Gly Lys Leu lie Ala Arg Ala Ala Arg Leu Val

340 345 350

Leu lie Ala Glu Glu Leu Asn Tyr Pro Asp Val lie Pro Lys Val Arg

355 360 365

Asn Phe Leu Lys Glu Thr I le Glu Pro Trp Leu Glu Gly Thr Phe Ser

370 375 380

Gly Asn Gly Phe Leu His Asp Glu Lys Trp Gly Gly lie He Thr Gin 0Z9 S19 019 ID S ^ I Li 丄 Λιο njo ID Ι ^ Λ 3 ^ IO niO SJV dsy nai B] y OJJ naq

909 009 969 jqi dJl nio 1¾Λ naq sA Ι¾Λ QUd dsy Ι¾Λ us _V J9 $ sqd 9Π υιν

06S S8S 089

nio J9S 811 ojj Eiv naq OJJ neq uio 911 ^ IO na Say SAQ nio

SAS OAS 99S.

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0 ^ SS9 OSS

Ti9i jqi Aio ^ IO "10 sAq Ι¾Λ SIH 丄 d i sAi J¾ d \\

Z 0Z SIS nio nai BIV -iqi nai J¾ JSS IO na ¾ιν Ι¾Λ OJJ ¾iv dsy io

019 SOS 009

ID siv ηθΐ λιο; ΘΜ Π8 ¾iv EIV ^^ S "^ 丄丄 ¾1V Ι ^ Λ ¾IV" ID

% QQV

J9S JUI J8S "ID UJO usy §JV ^ io dsy Jqi s nio jq _X naq AJO 3

08 S OL

¾IV dJ 丄 J9S SIH ^Η3 Ί Ι¾Λ "^ 丄 OJJ dsy si ^ SAQ §jy nsi S-iV 丄

09 0 usy J8S usy naq sAq J J 丄 dsy nsq usy na s dsy u |) eight sii jag

0

JA 丄 Biv uio OJJ sAi _ιλ 丄 sAi SJV IO cLi 丄 BfV OJJ dsy na sA 丄

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nai Ι ^ Λ ¾IV 811 ^ IO J ^ l 811 8Md ^ 10 ^S ! H 丄 s! H s! H dsy

Q \ V 0

usv J 911 ^ IO 3Md ^ IO 3 d dsy AJO _B1V dsy JRl JBS AJO SAI 00 S6C 06C S8C rO / 86df / lDd S908S / 86 O Phe Val Tyr Ala Leu Glu Gly Val Tyr Asp Asn Glu Ser Ala Leu Gin

625 630 635 640

Lys He Arg Asn Leu Lys Gly Phe Asp Gly Gly Asn Ser Leu Thr Asn

645 650 655

Leu Leu Trp Trp lie His Ser Arg Ser Asp Glu

660 665 SEQ ID NO: 4

Sequence length: 2291

Sequence type: nucleic acid

Number of chains: double strand

Topology: linear

Sequence type: cDNA to mRNA

Array features

Characteristic symbol: CDS

Location: 80..2080

Array:

CTTCTTTCCT CAACCTTCTT TCTTCTTATA TATTCGAACG ATCCGGGCCG CTCGAATTAA 60 ACCACCTTCA GCAACAATG GTT AAC ATC CAA ACC AAT ACA TCT TAC ATC TTC 112

Val Asn lie Gin Thr Asn Thr Ser Tyr lie Phe

1 5 10

CCT CAA ACA CAA TCC ACT GTT CTT CCT GAT CCC TCC AAA TTC TTC TCC 160 Pro Gin Thr Gin Ser Thr Val Leu Pro Asp Pro Ser Lys Phe Phe Ser

15 20 25

TCA AAC CTT CTC TCA AGT CCA CTC CCC ACA AAC TCT TTC TTC CAA AAC 208 Ser Asn Leu Leu Ser Ser Pro Leu Pro Thr Asn Ser Phe Phe Phe Gin Asn

30 35 40

TTT GTC CTA AAA AAT GGT GAC CAA CAA GAA TAC ATT CAT CCT TAC CTC 256 Phe Val Leu Lys Asn Gly Asp Gin Gin Glu Tyr lie His Pro Tyr Leu 45 50 55

ATC AAA TCC TCC AAC TCT TCC CTC TCT CTC TCA TAC CCT TCT CGC CAA 304 lie Lys Ser Ser Asn Ser Ser Leu Ser Leu Ser Tyr Pro Ser Arg Gin

60 65 70 75

GCC AGT TCA GCT GTC ATA TTC CAA GTC TTC AAT CCT GAT CTT ACC ATT 352 Ala Ser Ser Ala Val lie Phe Gin Val Phe Asn Pro Asp Leu Thr He

80 85 90

TCA GCC CCA CAA GGT CCC AAA CAA GGT CCC CCT GGT AAA CAC CTT ATC 400 Ser Ala Pro Gin Gly Pro Lys Gin Gly Pro Pro Gly Lys His Leu He

95 100 105

TCC TCC TAC AGT GAT CTC AGT GTC ACC TTG GAT TTC CCT TCT TCC AAT 448 Ser Ser Tyr Ser Asp Leu Ser Val Thr Leu Asp Phe Pro Ser Ser Asn

110 115 120

CTG AGC TTC TTC CTT GTT AGG GGA AGC CCC TAT TTG ACT GTG TCT GTG 496 Leu Ser Phe Phe Leu Val Arg Gly Ser Pro Tyr Leu Thr Val Ser Val

125 130 135

ACT CAA CCA ACT CCT CTT TCA ATT ACC ACC ATC CAT TCC ATT CTC TCA 544 Thr Gin Pro Thr Pro Leu Ser lie Thr Thr lie His Ser lie Leu Ser

140 145 150 155

TTC TCT TCA AAT GAC TCC AAC ACC AAG TAC ACC TTT CAG TTC AAC AAT 592 Phe Ser Ser Asn Asp Ser Asn Thr Lys Tyr Thr Phe Gin Phe Asn Asn

160 165 170

GGT CAA ACA TGG CTT CTT TAT GCT ACC TCC CCC ATC AAG TTG AAC CAC 640 Gly Gin Thr Trp Leu Leu Tyr Ala Thr Ser Prolie Lys Leu Asn His

175 180 185

ACC CTT TCT GAG ATA ACT TCT AAT GCA TTT TCT GGC ATA ATC CGG ATA 688 Thr Leu Ser Glu lie Thr Ser Asn Ala Phe Ser Gly lie lie Arg lie

190 195 200

GCT TTG TTG CCG GAT TCG GAT TCG AAA CAC GAG GCT GTT CTT GAC AAG 736 09ε 99S 09C

Ι¾Λ dsv OJd -ΐ usy nai njo njo s! V 811 nai naq Sjy ¾1V ¾1V

89Π DID IVO IDD OVl DVV Oil OVO OVO IDD 丄丄 V Oil VID DII ODV VDO 130

K OK S8C

SJV BIV 911 nsi sK- [K \ ^) aqj J9S nio jq jqi J¾ an J8S

0ΖΠ OOV VOO IIV Oil OVV 000 IVl 111 1VI VOl DVD VOV VDV I3V VIV VDl

J9S -I9S dsy nai J9S nio Ι¾Λ dsy sA JGS neq ¾iv J8S Ι¾Λ 911 01 VDl VDl IVD VIO ODV OVO 110 IVO VVV 131 110 330 V3I DID IIV OVO

9ΐε οτε οε οοε dsy SIH "ID nio sAq λιο sAq an SIH CLI 丄 J¾ Ι¾Λ sqd

^ OI IVO IVD DDI WO WD VVV DIV VOD OVV DIV VDl IVD DDI VOV VID 111

6Z 06Z S8Z naq OJd dsy J¾ sAq nai ΙΒΛ 丄 J9S dsy A \ Ι¾Λ ^ IO 1¾Λ

9 6 Oil 133 IVO VDV VVV Oil 110 001 VOl IVO 000 310 110 100 IID LL3

02Z LZ QLZ

dsy AJO dsy 811 J3S sAq ・ ΐΛ 丄 sA na dsy no ns ^ \ sAq dsy

8Z6 IVO 000 IVD UV OOV VVV IVl DVV V 丄丄 IVO WO 110 丄丄 V VVV OID IVO

99Z 092 Z

usy dsy usy §jy naq ng u]) eight SIH naq OJJ S! H ¾IV ns na

088 IVV 3V0 VOO IVV 103 113 113 OVO IID IVO DID IDD OVO 100 Oil 310

0 z 2n on

nsq ri9i dsy ^ ID ^J9 S dsy sA si ι> d_i 丄 usv ^J丄ⁿ I0 ί¾Λ sAo aqj VID DII IVD 000 VOl IVO VVV OVV OVO 001 OVV 1VI WO OID 101 Oil

OJd "ID SJV ³ ά Ι ^ Λ EIV sAi Ajo J8S Ι ^ Λ OJd 丄 SAQ S J9S 丄 8 O WD VOV Oil 010 130 VVV 100 V3 丄 DID 033 DVI 1DI 131 IOV IVl

SIZ 01Z SOZ sA dsy na eight ¾i v nio S! H SA ^ jag dsy J9S dsy OJJ neq naq ¾ | y I.Z0 / 86 <If / IDd S908S / 86 ATT CCA AAG GTT AGG AAT TTT TTG AAA GAA ACC ATT GAG CCA TGG TTG 1216 lie Pro Lys Val Arg Asn Phe Leu Lys Glu Thr I le Glu Pro Trp Leu

365 370 375

GAG GGA ACT TTT AGT GGG AAT GGA TTC CTA CAT GAT GAA AAA TGG GGT 1264 Glu Gly Thr Phe Ser Gly Asn Gly Phe Leu His Asp Glu Lys Trp Gly

380 385 390 395

GGC ATT ATT ACC CAA AAG GGG TCC ACT GAT GCT GGT GGT GAT TTT GGA 1312 Gly lie lie Thr Gin Lys Gly Ser Thr Asp Ala Gly Gly Asp Phe Gly

400 405 410

TTT GGA ATT TAC AAT GAT CAC CAC TAT CAT TTG GGG TAC TTC ATT TAT 1360 Phe Gly lie Tyr Asn Asp His His Tyr His Leu Gly Tyr Phe lie Tyr

415 420 425

GGA ATT GCG GTG CTC ACT AAG CTT GAT CCA GCA TGG GGT AGG AAG TAC 1408 Gly I le Ala Val Leu Thr Lys Leu Asp Pro Ala Trp Gly Arg Lys Tyr

430 435 440

AAG CCT CAA GCC TAT TCA ATA GTG CAA GAC TTC TTG AAC TTG GAC ACA 1456 Lys Pro Gin Ala Tyr Ser lie Val Gin Asp Phe Leu Asn Leu Asp Thr

445 450 455

AAA TTA AAC TCC AAT TAC ACA CGT TTG AGG TGT TTT GAC CCT TAT GTG 1504 Lys Leu Asn Ser Asn Tyr Thr Arg Leu Arg Cys Phe Asp Pro Tyr Val

460 465 470 475

CTT CAC TCT TGG GCT GGA GGG TTA ACT GAG TTC ACA GAT GGA AGG AAT 1552 Leu His Ser Trp 'Ala Gly Gly Leu Thr Glu Phe Thr Asp Gly Arg Asn

480 485 490

CAA GAG AGC ACA AGT GAG GCT GTG AGT GCA TAT TAT TCT GCT GCT TTG 1600 Gin Glu Ser Thr Ser Glu Ala Val Ser Ala Tyr Tyr Ser Ala Ala Leu

495 500 505

ATG GGA TTA GCA TAT GGT GAT GCA CCT CTT GTT GCA CTT GGA TCA ACA 1648 Met Gly Leu Ala Tyr Gly Asp Ala Pro Leu Val Ala Leu Gly Ser Thr 510 515 520

CTC ACA GCA TTG GAA ATT GAA GGG ACT AAA ATG TGG TGG CAT GTG AAA 1696 Leu Thr Ala Leu Glu .lie Glu Gly Thr Lys Met Trp Trp His Val Lys

525 530 535

GAG GGA GGT ACT TTG TAT GAG AAA GAG TTT ACA CAA GAG AAT AGG GTG 1744 Glu Gly Gly Thr Leu Tyr Glu Lys Glu Phe Thr Gin Glu Asn Arg Val

540 545 550 555

ATG GGT GTT CTA TGG TCT AAC AAG AGG GAC ACT GGA CTT TGG TTT GCT 1792 Met Gly Val Leu Trp Ser Asn Lys Arg Asp Thr Gly Leu Trp Phe Ala

560 565 570

CCT GCT GAG TGG AAA GAG TGT AGG CTT GGC ATT CAG CTC TTA CCA TTG 1840 Pro Ala Glu Trp Lys Glu Cys Arg Leu Gly lie Gin Leu Leu Pro Leu

575 580 585

GCT CCT ATT TCT GAA GCC ATT TTC TCC AAT GTT GAC TTT GTA AAG GAG 1888 Ala Pro lie Ser Glu Ala lie Phe Ser Asn Val Asp Phe Val Lys Glu

590 595 600

CTT GTG GAG TGG ACT TTG CCT GCT TTG GAT AGG GAG GGT GGT GTT GGT 1936 Leu Val Glu Trp Thr Leu Pro Ala Leu Asp Arg Glu Gly Gly Val Gly

605 610 615

GAA GGA TGG AAG GGG TTT GTG TAT GCC CTT GAA GGG GTT TAT GAC AAT 1984 Glu Gly Trp Lys Gly Phe Val Tyr Ala Leu Glu Gly Val Tyr Asp Asn

620 625 630 635

GAA AGT GCA CTG 'CAG AAG ATA AGA AAC CTG AAA GGT TTT GAT GGT GGA 2032 Glu Ser Ala Leu Gin Lys lie Arg Asn Leu Lys Gly Phe Asp Gly Gly

640 645 650

AAC TCT TTG ACC AAT CTC TTG TGG TGG ATT CAT AGC AGA AGT GAT GAA 2080 Asn Ser Leu Thr Asn Leu Leu Trp Trp lie His Ser Arg Ser Asp Glu

655 660 665

TAGAATGGAT TTGGTTATTG AACATGACAA CACTGCTGAT TTGGTTATTA TTATTATTGC 2140 TACTATTACT ATGCCAATAA AATCATCAAA TATGTCTTAC ATCATAATAA AAAAATTATT 2200 ATTATGCCAA TAAGTCAATG ATTTCGATAT GTTATAGAAG AAGAAAACTA TAATAAAGTT 2260 ACAACAATAT CATGATCTAT AACATGTCTG A 2291 SEQ ID NO: 5

Array length: 1054

Sequence type: amino acid

Topology: linear

Sequence type: protein

Array:

Met Asp Phe He Ser Ser Leu He Val Gly Cys Ala Gin Val Leu Cys

1 5 10 15

Glu Ser Met Asn Met Ala Glu Arg Arg Gly His Lys Thr Asp Leu Arg

20 25 30

Gin Ala He Thr Asp Leu Glu Thr Ala lie Gly Asp Leu Lys Ala lie

35 40 45

Arg Asp Asp Leu Thr Leu Arg lie Gin Gin Asp Gly Leu Glu Gly Arg

50 55 60

Ser Cys Ser Asn Arg Ala Arg Glu Trp Leu Ser Ala Val Gin Val Thr 65 70 75 80

Glu Thr Lys Thr Ala Leu Leu Leu Val Arg Phe Arg Arg Arg Glu Gin

85 90 95

Arg Thr Arg Met Arg Arg Arg Tyr Leu Ser Cys Phe Gly Cys Ala Asp

100 105 110

Tyr Lys Leu Cys Lys Lys Val Ser Ala He Leu Lys Ser lie Gly Glu

115 120 125

Leu Arg Glu Arg Ser Glu Ala lie Lys Thr Asp Gly Gly Ser lie Gin

130 135 140

Val Thr Cys Arg Glu lie Pro lie Lys Ser Val Val Gly Asn Thr Thr 145 150 155 160

Met Met Glu Gin Val Leu Glu Phe Leu Ser Glu Glu Glu Glu Arg Gly

165 170 175 lie lie Gly Val Tyr Gly Pro Gly Gly Val Gly Lys Thr Thr Leu Met

180 185 190

Gin Ser lie Asn Asn Glu Leu lie Thr Lys Gly His Gin Tyr Asp Val

195 200 205

Leu lie Trp Val Gin Met Ser Arg Glu Phe Gly Glu Cys Thr lie Gin

210 215 220

Gin Ala Val Gly Ala Arg Leu Gly Leu Ser Trp Asp Glu Lys Glu Thr 225 230 235 240

Gly Glu Asn Arg Ala Leu Lys lie Tyr Arg Ala Leu Arg Gin Lys Arg

245 250 255

Phe Leu Leu Leu Leu Asp Asp Val Trp Glu Glu lie Asp Leu Glu Lys

260 265 270

Thr Gly Val Pro Arg Pro Asp Arg Glu Asn Lys Cys Lys Val Met Phe

275 280 285

Thr Thr Arg Ser lie Ala Leu Cys Asn Asn Met Gly Ala Glu Tyr Lys

290 295 300

Leu Arg Val Glu Phe Leu Glu Lys Lys His Ala Trp Glu Leu Phe Cys 305 310 315 320

Ser Lys Val Trp Arg Lys Asp Leu Leu Glu Ser Ser Ser lie Arg Arg

.325 330 335

Leu Ala Glu lie lie Val Ser Lys Cys Gly Gly Leu Pro Leu Ala Leu

340 345 350 lie Thr Leu Gly Gly Ala Met Ala Gly Leu Gin Glu Phe Asp Pro Gly

355 360 365

Ser Pro Gly Leu Gin Glu Phe Ala Ala Ala Arg lie Lys Pro Pro Ser 370 375 380 Ala Thr Met Val Asn He Gin Thr Asn Thr Ser Tyr lie Phe Pro Gin 385 390 395 400

Thr Gin Ser Thr Val Leu Pro Asp Pro Ser Lys Phe Phe Ser Ser Asn

405 410 415

Leu Leu Ser Ser Pro Leu Pro Thr Asn Ser Phe Phe Gin Asn Phe Val

420 425 430

Leu Lys Asn Gly Asp Gin Gin Glu Tyr lie His Pro Tyr Leu lie Lys

435 440 445

Ser Ser Asn Ser Ser Leu Ser Leu Ser Tyr Pro Ser Arg Gin Ala Ser

450 455 460

Ser Ala Val He Phe Gin Val Phe Asn Pro Asp Leu Thr lie Ser Ala 465 470 475 480

Pro Gin Gly Pro Lys Gin Gly Pro Pro Gly Lys His Leu lie Ser Ser

485 490 495

Tyr Ser Asp Leu Ser Val Thr Leu Asp Phe Pro Ser Ser Asn Leu Ser

500 505 510

Phe Phe Leu Val Arg Gly Ser Pro Tyr Leu Thr Val Ser Val Thr Gin

515 520 525

Pro Thr Pro Leu Ser lie Thr Thr lie His Ser I le Leu Ser Phe Ser

530 535 540

Ser Asn Asp Ser Asn Thr Lys Tyr Thr Phe Gin Phe Asn Asn Gly Gin 545 550 555 560

Thr Trp Leu Leu Tyr Ala Thr Ser Pro lie Lys Leu Asn His Thr Leu

565 570 575

Ser Glu He Thr Ser Asn Ala Phe Ser Gly lie lie Arg lie Ala Leu

580 585 590

Leu Pro Asp Ser Asp Ser Lys His Glu Ala Val Leu Asp Lys Tyr Ser

595 600 605

Ser Cys Tyr Pro Val Ser Gly Lys Ala Val Phe Arg Glu Pro Phe Cys 610 615 620

Val Glu Tyr Asn Trp Glu Lys Lys Asp Ser Gly Asp Leu Leu Leu Leu 625 630 635 640

Ala His Pro Leu His Val Gin Leu Leu Arg Asn Gly Asp Asn Asp Val

645 650 655

Lys lie Leu Glu Asp leu Lys Tyr Lys Ser lie Asp Gly Asp Leu Val

660 665 670

Gly Val Val Gly Asp Ser Trp Val Leu Lys Thr Asp Pro Leu Phe Val

675 680 685

Thr Trp His Ser He Lys Gly lie Lys Glu Glu Ser His Asp Glu lie

690 695 700

Val Ser Ala Leu Ser Lys Asp Val Glu Ser Leu Asp Ser Ser Ser lie 705 710 715 720

Thr Thr Thr Glu Ser Tyr Phe Tyr Gly Lys Leu lie Ala Arg Ala Ala

725 730 735

Arg Leu Val Leu lie Ala Glu Glu Leu Asn Tyr Pro Asp Val lie Pro

740 745 750

Lys Val Arg Asn Phe Leu Lys Glu Thr He Glu Pro Trp Leu Glu Gly

755 760 765

Thr Phe Ser Gly Asn Gly Phe Leu His Asp Glu Lys Trp Gly Gly He

770 775 780

lie Thr Gin Lys Gly Ser Thr Asp Ala Gly Gly Asp Phe Gly Phe Gly 785 790 795 800

He Tyr Asn Asp His His Tyr His Leu Gly Tyr Phe He Tyr Gly lie

805 810 815

Ala Val Leu Thr Lys Leu Asp Pro Ala Trp Gly Arg Lys Tyr Lys Pro

820 825 830

Gin Ala Tyr Ser lie Val Gin Asp Phe Leu Asn Leu Asp Thr Lys Leu

835 840 845 Asn Ser Asn Tyr Thr Arg Leu Arg Cys Phe Asp Pro Tyr Val Leu His

850 855 860

Ser Trp Ala Gly Gly Leu Thr Glu Phe Thr Asp Gly Arg Asn Gin Glu 865 870 875 880

Ser Thr Ser Glu Ala Val Ser Ala Tyr Tyr Ser Ala Ala Leu Met Gly

885 890 895

Leu Ala Tyr Gly Asp Ala Pro Leu Val Ala Leu Gly Ser Thr Leu Thr

900 905 910

Ala Leu Glu lie Glu Gly Thr Lys Met Trp Trp His Val Lys Glu Gly

915 920 925

Gly Thr Leu Tyr Glu Lys Glu Phe Thr Gin Glu Asn Arg Val Met Gly

930 935 940

Val Leu Trp Ser Asn Lys Arg Asp Thr Gly Leu Trp Phe Ala Pro Ala 945 950 955 960

Glu Trp Lys Glu Cys Arg Leu Gly lie Gin Leu Leu Pro Leu Ala Pro

965 970 975 lie Ser Glu Ala He Phe Ser Asn Val Asp Phe Val Lys Glu Leu Val

980 985 990

Glu Trp Thr Leu Pro Ala Leu Asp Arg Glu Gly Gly Val Gly Glu Gly

995 1000 1005

Trp Lys Gly Phe Val Tyr Ala Leu Glu Gly Val Tyr Asp Asn Glu Ser

1010 1015 1020

Ala Leu Gin Lys lie Arg Asn Leu Lys Gly Phe Asp Gly Gly Asn Ser 1025 1030 1035 1040

Leu Thr Asn Leu Leu Trp Trp lie His Ser Arg Ser Asp Glu

1045 1050 SEQ ID NO: 6

Sequence length: 3405 Sequence type: nucleic acid

Number of chains: double strand

Topology: linear

Sequence type: cDNA to mRNA

Array features

Characteristic symbol: CDS

Location: 33..3194

Array:

ACAAGTAAAA GAAAGAGCGA GAAATCATCG AA ATG GAT TTC ATC TCA TCT CTT 53

Met Asp Phe lie Ser Ser Leu

1 5

ATC GTT GGC TGT GCT CAG GTG TTG TGT GAA TCT ATG AAT ATG GCG GAG 101 lie Val Gly Cys Ala Gin Val Leu Cys Glu Ser Met Asn Met Ala Glu

10 15 20

AGA AGA GGA CAT AAG ACT GAT CTT AGA CAA GCC ATC ACT GAT CTT GAA 149 Arg Arg Gly His Lys Thr Asp Leu Arg Gin Ala lie Thr Asp Leu Glu

25 30 35

ACA GCC ATC GGT GAC TTG AAG GCC ATA CGT GAT GAC CTG ACT TTA CGG 197 Thr Ala lie Gly Asp Leu Lys Ala lie Arg Asp Asp Leu Thr Leu Arg

40 45 50 55

ATC CAA CAA GAC GGT CTA GAG GGA CGA AGC TGC TCA AAT CGT GCC AGA 245 lie Gin Gin Asp Gly Leu Glu Gly Arg Ser Cys Ser Asn Arg Ala Arg

60 65 70

GAG TGG CTT AGT GCG GTG CAA GTA ACG GAG ACT AAA ACA GCC CTA CTT 293 Glu Trp Leu Ser Ala Val Gin Val Thr Glu Thr Lys Thr Ala Leu Leu

75 80 85

TTA GTG AGG TTT AGG CGT CGG GAA CAG AGG ACG CGA ATG AGG AGG AGA 341 Leu Val Arg Phe Arg Arg Arg Glu Gin Arg Thr Arg Met Arg Arg Arg

90 95 100 TAC CTC AGT TGT TTC GGT TGT GCC GAC TAC AAA CTG TGC AAG AAG GTT 389 Tyr Leu Ser Cys Phe Gly Cys Ala Asp Tyr Lys Leu Cys Lys Lys Val

105 110 115

TCT GCC ATA TTG AAG AGC ATT GGT GAG CTG AGA GAA CGC TCT GAA GCT 437 Ser Ala lie Leu Lys Ser lie Gly Glu Leu Arg Glu Arg Ser Glu Ala

120 125 130 135

ATC AAA ACA GAT GGC GGG TCA ATT CAA GTA ACT TGT AGA GAG ATA CCC 485 lie Lys Thr Asp Gly Gly Ser We Gin Val Thr Cys Arg Glu He Pro

140 145 150

ATC AAG TCC GTT GTC GGA AAT ACC ACG ATG ATG GAA CAG GTT TTG GAA 533 lie Lys Ser Val Val Gly Asn Thr Thr Met Met Glu Gin Val Leu Glu

155 160 165

TTT CTC AGT GAA GAA GAA GAA AGA GGA ATC ATT GGT GTT TAT GGA CCT 581 Phe Leu Ser Glu Glu Glu Glu Arg Gly He lie Gly Val Tyr Gly Pro

170 175 180

GGT GGG GTT GGG AAG ACA ACG TTA ATG CAG AGC ATT AAC AAC GAG CTG 629 Gly Gly Val Gly Lys Thr Thr Leu Met Gin Ser lie Asn Asn Glu Leu

185 190 195

ATC ACA AAA GGA CAT CAG TAT GAT GTA CTG ATT TGG GTT CAA ATG TCC 677 lie Thr Lys Gly His Gin Tyr Asp Val Leu lie Trp Val Gin Met Ser

200 205 210 215

AGA GAA TTC GGC GAG TGT ACA ATT CAG CAA GCC GTT GGA GCA CGG TTG 725 Arg Glu Phe Gly Glu Cys Thr He Gin Gin Ala Val Gly Ala Arg Leu

220 225 230

GGT TTA TCT TGG GAC GAG AAG GAG ACC GGC GAA AAC AGA GCT TTG AAG 773 Gly Leu Ser Trp Asp Glu Lys Glu Thr Gly Glu Asn Arg Ala Leu Lys

235 240 245

ATA TAC AGA GCT TTG AGA CAG AAA CGT TTC TTG TTG TTG CTA GAT GAT 821 lie Tyr Arg Ala Leu Arg Gin Lys Arg Phe Leu Leu Leu Leu Asp Asp 250 255 260

GTC TGG GAA GAG ATA GAC TTG GAG AAA ACT GGA GTT CCT CGA CCT GAC 869 Val Trp Glu Glu lie Asp Leu Glu Lys Thr Gly Val Pro Arg Pro Asp

265 270 275

AGG GAA AAC AAA TGC AAG GTG ATG TTC ACG ACA CGG TCT ATA GCA TTA 917 Arg Glu Asn Lys Cys Lys Val Met Phe Thr Thr Arg Ser lie Ala Leu

280 285 290 295

TGC AAC AAT ATG GGT GCG GAA TAC AAG TTG AGA GTG GAG TTT CTG GAG 965 Cys Asn Asn Met Gly Ala Glu Tyr Lys Leu Arg Val Glu Phe Leu Glu

300 305 310

AAG AAA CAC GCG TGG GAG CTG TTC TGT AGT AAG GTA TGG AGA AAA GAT 1013 Lys Lys His Ala Trp Glu Leu Phe Cys Ser Lys Val Trp Arg Lys Asp

315 320 325

CTT TTA GAG TCA TCA TCA ATT CGC CGG CTC GCG GAG ATT ATA GTG AGT 1061 Leu Leu Glu Ser Ser Ser lie Arg Arg Leu Ala Glu I le lie Val Ser

330 335 340

AAA TGT GGA GGA TTG CCA CTA GCG TTG ATC ACT TTA GGA GGA GCC ATG 1109 Lys Cys Gly Gly Gly Leu Pro Leu Ala Leu lie Thr Leu Gly Gly Ala Met

345 350 355

GCT GGG CTG CAG GAA TTC GAT CCC GGA TCC CCC GGG CTG CAG GAA TTC 1157 Ala Gly Leu Gin Glu Phe Asp Pro Gly Ser Pro Gly Leu Gin Glu Phe

360 365 370 375

GCG GCC GCT CGA ATT AAA CCA CCT TCA GCA ACA ATG GTT AAC ATC CAA 1205 Ala Ala Ala Arg lie Lys Pro Pro Ser Ala Thr Met Val Asn He Gin

380 385 390

ACC AAT ACA TCT TAC ATC TTC CCT CAA ACA CAA TCC ACT GTT CTT CCT 1253 Thr Asn Thr Ser Tyr lie Phe Pro Gin Thr Gin Ser Thr Val Leu Pro

395 400 405

GAT CCC TCC AAA TTC TTC TCC TCA AAC CTT CTC TCA AGT CCA CTC CCC 1301 Asp Pro Ser Lys Phe Phe Ser Ser Asn Leu Leu Ser Ser Pro Leu Pro

410 415 420

ACA AAC TCT TTC TTC CAA AAC TTT GTC CTA AAA AAT GGT GAC CAA CAA 1349 Thr Asn Ser Phe Phe Gin Asn Phe Val Leu Lys Asn Gly Asp Gin Gin

425 430 435

GAA TAC ATT CAT CAT CCT TAC CTC ATC AAA TCC TCC AAC TCT TCC CTC TCT 1397 Glu Tyr lie His Pro Tyr Leu lie Lys Ser Ser Asn Ser Ser Leu Ser

440 445 450 455

CTC TCA TAC CCT TCT CGC CAA GCC AGT TCA GCT GTC ATA TTC CAA GTC 1445 Leu Ser Tyr Pro Ser Arg Gin A 1 a Ser Ser Ala Val lie Phe Gin Val

460 465 470

TTC AAT CCT GAT CTT ACC ATT TCA GCC CCA CAA GGT CCC AAA CAA GGT 1493 Phe Asn Pro Asp Leu Thr lie Ser Ala Pro Gin Gly Pro Lys Gin Gly

475 480 485

CCC CCT GGT AAA CAC CTT ATC TCC TCC TAC AGT GAT CTC AGT GTC ACC 1541 Pro Pro Gly Lys His Leu lie Ser Ser Tyr Ser Asp Leu Ser Val Thr

490 495 500

TTG GAT TTC CCT TCT TCC AAT CTG AGC TTC TTC CTT GTT AGG GGA AGC 1589 Leu Asp Phe Pro Ser Ser Asn Leu Ser Phe Phe Leu Val Arg Gly Ser

505 510 515

CCC TAT TTG ACT GTG TCT GTG ACT CAA CCA ACT CCT CTT TCA ATT ACC 1637 Pro Tyr Leu Thr Val Ser Val Thr Gin Pro Thr Pro Leu Serlie Thr

520 525 530 535

ACC ATC CAT TCC ATT CTC TCA TTC TCT TCA AAT GAC TCC AAC ACC AAG 1685 Thr lie His Ser lie Leu Ser Phe Ser Ser Asn Asp Ser Asn Thr Lys

540 545 550

TAC ACC TTT CAG TTC AAC AAT GGT CAA ACA TGG CTT CTT TAT GCT ACC 1733 Tyr Thr Phe Gin Phe Asn Asn Gly Gin Thr Trp Leu Leu Tyr Ala Thr

555 560 565 TCC CCC ATC AAG TTG AAC CAC ACC CTT TCT GAG ATA ACT TCT AAT GCA 1781 Ser Pro lie Lys Leu Asn His Thr Leu Ser Glu lie Thr Ser Asn Ala

570 575 580

TTT TCT GGC ATA ATC CGG ATA GCT TTG TTG CCG GAT TCG GAT TCG AAA 1829 Phe Ser Gly lie lie Arg lie Ala Leu Leu Pro Asp Ser Asp Ser Lys

585 590 595

CAC GAG GCT GTT CTT GAC AAG TAT AGT TCT TGT TAC CCC GTG TCA GGT 1877 His Glu Ala Val Leu Asp Lys Tyr Ser Ser Cys Tyr Pro Val Ser Gly

600 605 610 615

AAA GCT GTG TTC AGA GAA CCT TTC TGT GTG GAA TAT AAC TGG GAG AAG 1925 Lys Ala Val Phe Arg Glu Pro Phe Cys Val Glu Tyr Asn Trp Glu Lys

620 625 630

AAA GAT TCA GGG GAT TTG CTA CTC TTG GCT CAC CCT CTC CAT GTT CAG 1973 Lys Asp Ser Gly Asp Leu Leu Leu Leu Ala His Pro Leu His Val Gin

635 640 645

CTT CTT CGT AAT GGA GAC AAT GAT GTC AAA ATT CTT GAA GAT TTA AAG 2021 Leu Leu Arg Asn Gly Asp Asn Asp Val Lys lie Leu Glu Asp Leu Lys

650 655 660

TAT AAA AGC ATT GAT GGG GAT CTT GTT GGT GTT GTC GGG GAT TCA TGG 2069 Tyr Lys Ser lie Asp Gly Asp Leu Val Gly Val Val Gly Asp Ser Trp

665 670 675

GTT TTG AAA ACA GAT CCT TTG TTT GTA ACA TGG CAT TCA ATC AAG GGA 2117 Val Leu Lys Thr Asp Pro Leu Phe Val Thr Trp His Serlie Lys Gly

680 685 690 695

ATC AAA GAA GAA TCC CAT GAT GAG ATT GTC TCA GCC CTT TCT AAA GAT 2165 lie Lys Glu Glu Ser His Asp Glu lie Val Ser Ala Leu Ser Lys Asp

700 705 710

GTT GAG AGC CTA GAT TCA TCA TCA ATA ACT ACA ACA GAG TCA TAT TTT 2213 Val Glu Ser Leu Asp Ser Ser Serlie Thr Thr Thr Glu Ser Tyr Phe 715 720 725

TAT GGG AAG TTG ATT GCA AGG GCT GCA AGG TTG GTA TTG ATT GCT GAG 2261 Tyr Gly Lys Leu lie Ala Arg Ala Ala Arg Leu Val Leu lie Ala Glu

730 735 740

GAG TTG AAC TAC CCT GAT GTG ATT CCA AAG GTT AGG AAT TTT TTG AAA 2309 Glu Leu Asn Tyr Pro Asp Val lie Pro Lys Val Arg Asn Phe Leu Lys

745 750 755

GAA ACC ATT GAG CCA TGG TTG GAG GGA ACT TTT AGT GGG AAT GGA TTC 2357 Glu Thr lie Glu Pro Trp Leu Glu Gly Thr Phe Ser Gly Asn Gly Phe

760 765 770 775

CTA CAT GAT GAA AAA TGG GGT GGC ATT ATT ACC CAA AAG GGG TCC ACT 2405 Leu His Asp Glu Lys Trp Gly Gly lie lie Thr Gin Lys Gly Ser Thr

780 785 790

GAT GCT GGT GGT GAT TTT GGA TTT GGA ATT TAC AAT GAT CAC CAC TAT 2453 Asp Ala Gly Gly Asp Phe Gly Phe Glylie Tyr Asn Asp His His Tyr

795 800 805

CAT TTG GGG TAC TTC ATT TAT GGA ATT GCG GTG CTC ACT AAG CTT GAT 2501 His Leu Gly Tyr Phe lie Tyr Gly lie Ala Val Leu Thr Lys Leu Asp

810 815 820

CCA GCA TGG GGT AGG AAG TAC AAG CCT CAA GCC TAT TCA ATA GTG CAA 2549 Pro Ala Trp Gly Arg Lys Tyr Lys Pro Gin Ala Tyr Ser lie Val Gin

825 830 835

GAC TTC TTG AAC TTG GAC ACA AAA TTA AAC TCC AAT TAC ACA CGT TTG 2597 Asp Phe Leu Asn Leu Asp Thr Lys Leu Asn Ser Asn Tyr Thr Arg Leu

840 845 850 855

AGG TGT TTT GAC CCT TAT GTG CTT CAC TCT TGG GCT GGA GGG TTA ACT 2645 Arg Cys Phe Asp Pro Tyr Val Leu His Ser Trp Ala Gly Gly Leu Thr

860 865 870

GAG TTC ACA GAT GGA AGG AAT CAA GAG AGC ACA AGT GAG GCT GTG AGT 2693 Glu Phe Thr Asp Gly Arg Asn Gin Glu Ser Thr Ser Glu Ala Val Ser

875 880 885

GCA TAT TAT TCT GCT GCT TTG ATG GGA TTA GCA TAT GGT GAT GCA CCT 2741 Ala Tyr Tyr Ser Ala Ala Leu Met Gly Leu Ala Tyr Gly Asp Ala Pro

890 895 900

CTT GTT GCA CTT GGA TCA ACA CTC ACA GCA TTG GAA ATT GAA GGG ACT 2789 Leu Val Ala Leu Gly Ser Thr Leu Thr Ala Leu Glulie Glu Gly Thr

905 910 915

AAA ATG TGG TGG CAT GTG AAA GAG GGA GGT ACT TTG TAT GAG AAA GAG 2837 Lys Met Trp Trp His Val Lys Glu Gly Gly Thr Leu Tyr Glu Lys Glu

920 925 930 935

TTT ACA CAA GAG AAT AGG GTG ATG GGT GTT CTA TGG TCT AAC AAG AGG 2885 Phe Thr Gin Glu Asn Arg Val Met Gly Val Leu Trp Ser Asn Lys Arg

940 945 950

GAC ACT GGA CTT TGG TTT GCT CCT GCT GAG TGG AAA GAG TGT AGG CTT 2933 Asp Thr Gly Leu Trp Phe Ala Pro Ala Glu Trp Lys Glu Cys Arg Leu

955 960 965

GGC ATT CAG CTC TTA CCA TTG GCT CCT ATT TCT GAA GCC ATT TTC TCC 2981 Gly He Gin Leu Leu Pro Leu Ala Pro lie Ser Glu Ala lie Phe Ser

970 975 980

AAT GTT GAC TTT GTA AAG GAG CTT GTG GAG TGG ACT TTG CCT GCT TTG 3029 Asn Val Asp Phe Val Lys Glu Leu Val Glu Trp Thr Leu Pro Ala Leu

985 990 995

GAT AGG GAG GGT GGT GTT GGT GAA GGA TGG AAG GGG TTT GTG TAT GCC 3077 Asp Arg Glu Gly Gly Val Gly Glu Gly Trp Lys Gly Phe Val Tyr Ala

1000 1005 1010 1015

CTT GAA GGG GTT TAT GAC AAT GAA AGT GCA CTG CAG AAG ATA AGA AAC 3125 Leu Glu Gly Val Tyr Asp Asn Glu Ser Ala Leu Gin Lys lie Arg Asn

1020 1025 1030 CTG AAA GGT TTT GAT GGT GGA AAC TCT TTG ACC AAT CTC TTG TGG TGG 3173 Leu Lys Gly Phe Asp Gly Gly Asn Ser Leu Thr Asn Leu Leu Trp Trp

1035 1040 1045

ATT CAT AGC AGA AGT GAT GAA TAGAATGGAT TTGGTTATTG AACATGACAA 3224 lie His Ser Arg Ser Asp Glu

1050

CACTGCTGAT TTGGTTATTA TTATTATTGC TACTATTACT ATGCCAATAA AATCATCAAA 3284 TATGTCTTAC ATCATAATAA AAAAATTATT ATTATGCCAA TAAGTCAATG ATTTCGATAT 3344 GTTATAGAAG AAGAAAACTA TAATAAAGTT ACAACAATAT CATGATCTAT AACATGTCTG 3404

Sequence length: 877

Sequence type: amino acid

Topology: linear

Sequence type: protein

Array:

Val Asn lie Gin Thr Asn Thr Ser Tyr lie Phe Pro Gin Thr Gin Ser

1 5 10 15

Thr Val Leu Pro Asp Pro Ser Lys Phe Phe Ser Ser Asn Leu Leu Ser

20 25 30

Ser Pro Leu Pro Thr Asn Ser Phe Phe Gin Asn Phe Val Leu Lys Asn

3540 45

Gly Asp Gin Gin Glu Tyr lie His Pro Tyr Leu lie Lys Ser Ser Asn

50 55 60

Ser Ser Leu Ser Leu Ser Tyr Pro Ser Arg Gin Ala Ser Ser Ala Val 65 70 75 80

He Phe Gin Val Phe Asn Pro Asp Leu Thr I le Ser Ala Pro Gin Gly

85 90 95 Pro Lys Gin Gly Pro Pro Gly Lys His Leu 11 e Ser Ser Tyr Ser Asp

100 105 110

Leu Ser Val Thr Leu Asp Phe Pro Ser Ser Asn Leu Ser Phe Phe Leu

115 120 125

Val Arg Gly Ser Pro Tyr Leu Thr Val Ser Val Thr Gin Pro Thr Pro

130 135 140

Leu Ser lie Thr Thr lie His Ser lie Leu Ser Phe Ser Ser Asn Asp 145 150 155 160

Ser Asn Thr Lys Tyr Thr Phe Gin Phe Asn Asn Gly Gin Thr Trp Leu

165 170 175

Leu Tyr Ala Thr Ser Pro I le Lys Leu Asn His Thr Leu Ser Glu I le

180 185 190

Thr Ser Asn Ala Phe Ser Gly lie lie Arg I le Ala Leu Leu Pro Asp

195 200 205

Ser Asp Ser Lys His Glu Ala Val Leu Asp Lys Tyr Ser Ser Cys Tyr

210 215 220

Pro Val Ser Gly Lys Ala Val Phe Arg Glu Pro Phe Cys Val Glu Tyr 225 230 235 240

Asn Trp Glu Lys Lys Asp Ser Gly Asp Leu Leu Leu Leu Ala His Pro

245 250 255

Leu His Val Gin Leu Leu Arg Asn Gly Asp Asn Asp Val Lys lie Leu

260 265 270

Glu Asp Leu Lys Tyr Lys Ser lie Asp Gly Asp Leu Val Gly Val Val

275 280 285

Gly Asp Ser Trp Val Leu Lys Thr Asp Pro Leu Phe Val Thr Trp His

290 295 300

Ser lie Lys Gly He Lys Glu Glu Ser His Asp Glu lie Val Ser Ala 305 310 315 320

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565 570 575

Glu Thr Ala lie Gly Asp Leu Lys Ala lie Arg Asp Asp Leu Thr Leu

580 585 590

Arg lie Gin Gin Asp Gly Leu Glu Gly Arg Ser Cys Ser Asn Arg Ala

595 600 605

Arg Glu Trp Leu Ser Ala Val Gin Val Thr Glu Thr Lys Thr Ala Leu

610 615 620

Leu Leu Val Arg Phe Arg Arg Arg Glu Gin Arg Thr Arg Met Arg Arg 625 630 635 640

Arg Tyr Leu Ser Cys Phe Gly Cys Ala Asp Tyr Lys Leu Cys Lys Lys

645 650 655

Val Ser Ala lie Leu Lys Ser lie Gly Glu Leu Arg Glu Arg Ser Glu

660 665 670

Ala lie Lys Thr Asp Gly Gly Ser lie Gin Val Thr Cys Arg Glu lie

675 680 685

Pro lie Lys Ser Val Val Gly Asn Thr Thr Met Met Glu Gin Val Leu

690 695 700

Glu Phe Leu Ser Glu Glu Glu Glu Arg Gly lie lie Gly Val Tyr Gly 705 710 715 720

Pro Gly Gly Val Gly Lys Thr Thr Leu Met Gin Ser lie Asn Asn Glu

725 730 735

Leu lie Thr Lys Gly His Gin Tyr Asp Val Leu lie Trp Val Gin Met

740 745 750

Ser Arg Glu Phe Gly Glu Cys Thr He Gin Gin Ala Val Gly Ala Arg

755 760 765

Leu Gly Leu Ser Trp Asp Glu Lys Glu Thr Gly Glu Asn Arg Ala Leu

770 775 780

Lys lie Tyr Arg Ala Leu Arg Gin Lys Arg Phe Leu Leu Leu Leu Asp 785 790 795 800

Asp Val Trp Glu Glu lie Asp Leu Glu Lys Thr Gly Val Pro Arg Pro

805 810 815

Asp Arg Glu Asn Lys Cys Lys Val Met Phe Thr Thr Arg Ser lie Ala

820 825 830

Leu Cys Asn Asn Met Gly Ala Glu Tyr Lys Leu Arg Val Glu Phe Leu

835 840 845

Glu Lys Lys His Ala Trp Glu Leu Phe Cys Ser Lys Val Trp Arg Lys

850 855 860

Asp Leu Leu Glu Ser Ser Ser lie Arg Arg Leu Ala Glu

865 870 875 SEQ ID NO: 8

Array length: 2666

Sequence type: nucleic acid

Number of chains: double strand

Topology: linear

Sequence type: cDNA to mRNA

Array features

Characteristic symbol: CDS

Location: 29..2659

Array:

TCGAATTAAA CGACCTTCAG CAACAATG GTT AAC ATC CAA ACC AAT ACA TCT 52

Val Asn lie Gin Thr Asn Thr Ser

1 5

10 15 20

25 30 35 40

45 50 55

CCT TAC CTC ATC AAA TCC TCC AAC TCT TCC CTC TCT CTC TCA TAC CCT 244 Pro Tyr Leu ly Lys Ser Ser Asn Ser Ser Leu Ser Leu Ser Tyr Pro

60 65 70

75 80 85

90 95 100

105 110 115 120

125 130 135

GTG TCT GTG ACT CAA CCA ACT CCT CTT TCA ATT ACC ACC ATC CAT TCC 484 Val Ser Val Thr Gin Pro Thr Pro Leu Ser He Thr Thr lie His Ser

1.40 145 150

ATT CTC TCA TTC TCT TCA AAT GAC TCC AAC ACC AAG TAC ACC TTT CAG 532 He Leu Ser Phe Ser Ser Asn Asp Ser Asn Thr Lys Tyr Thr Phe Gin

155 160 165

170 175 180 TTG AAC CAC ACC CTT TCT GAG ATA ACT TCT AAT GCA TTT TCT GGC ATA 628

Leu Asn His Thr Leu Ser Glu lie Thr Ser Asn Ala Phe Ser Gly lie

185 190 195 200

205 210 215

220 225 230

235 240 245

250 255 260

GGA GAC AAT GAT GTC AAA ATT CTT GAA GAT TTA AAG TAT AAA AGC ATT 868 Gly Asp Asn Asp Val Lys lie Leu Glu Asp Leu Lys Tyr Lys Ser He

265 270 275 280

285 290 295

GAT CCT TTG TTT GTA ACA TGG CAT TCA ATC AAG GGA ATC AAA GAA GAA 964 Asp Pro Leu Phe Val Thr Trp His Ser He Lys Gly lie Lys Glu Glu

300 305 310

315 320 325

GAT TCA TCA TCA ATA ACT ACA ACA GAG TCA TAT TTT TAT GGG AAG TTG 1060 Asp Ser Ser Ser He Thr Thr Thr Glu Ser Tyr Phe Tyr Gly Lys Leu 0½I 131 IVl IVl VDO IOV DID IDD OVD IOV VOV 30V OVO VVD IVV OOV VDO

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8011 OVl DVV Oil OVO OVO IDD 11V Oil VIO DII ODV VOD 13D OOV VOO 11V

on sec οεε

6 Z0 / 86df / X3d S908S / 86 OM Gly Arg Asn Gin Glu Ser Thr Ser Glu Ala Val Ser A 1 a Tyr Tyr Ser

490 495 500

505 510 515 520

GGA TCA ACA CTC ACA GCA TTG GAA ATT GAA GGG ACT AAA ATG TGG TGG 1636 Gly Ser Thr Leu Thr Ala Leu Glulie Glu Gly Thr Lys Met Trp Trp

525 530 535

CAT GCG ATG GAT TTC ATC TCA TCT CTT ATC GTT GGC TGT GCT CAG GTG 1684 His Ala Met Asp Phe lie Ser Ser Leu lie Val Gly Cys Ala Gin Val

540 545 550

TTG TGT GAA TCT ATG AAT ATG GCG GAG AGA AGA GGA CAT AAG ACT GAT 1732 Leu Cys Glu Ser Met Asn Met Ala Glu Arg Arg Gly His Lys Thr Asp

555 560 565

CTT AGA CAA GCC ATC ACT GAT CTT GAA ACA GCC ATC GGT GAC TTG AAG 1780 Leu Arg Gin Ala lie Thr Asp Leu Glu Thr Ala lie Gly Asp Leu Lys

570 575 580

GCC ATA CGT GAT GAC CTG ACT TTA CGG ATC CAA CAA GAC GGT CTA GAG 1828 Ala lie Arg Asp Asp Leu Thr Leu Arg lie Gin Gin Asp Gly Leu Glu

585 590 595 600

GGA CGA AGC TGC TCA AAT CGT GCC AGA GAG TGG CTT AGT GCG GTG CAA 1876 Gly Arg Ser Cys Ser Asn Arg Ala Arg Glu Trp Leu Ser Ala Val Gin

605 610 615

GTA ACG GAG ACT AAA ACA GCC CTA CTT TTA GTG AGG TTT AGG CGT CGG 1924 Val Thr Glu Thr Lys Thr Ala Leu Leu Leu Val Arg Phe Arg Arg Arg

620 625 630

GAA CAG AGG ACG CGA ATG AGG AGG AGA TAC CTC AGT TGT TTC GGT TGT 1972 Glu Gin Arg Thr Arg Met Arg Arg Arg Tyr Leu Ser Cys Phe Gly Cys

635 640 645 GCC GAC TAC AAA CTG TGC AAG AAG GTT TCT GCC ATA TTG AAG AGC ATT 2020 Ala Asp Tyr Lys Leu Cys Lys Lys Val Ser Ala lie Leu Lys Ser lie

650 '655 660

GGT GAG CTG AGA GAA CGC TCT GAA GCT ATC AAA ACA GAT GGC GGG TCA 2068 Gly Glu Leu Arg Glu Arg Ser Glu Ala lie Lys Thr Asp Gly Gly Ser

665 670 675 680

ATT CAA GTA ACT TGT AGA GAG ATA CCC ATC AAG TCC GTT GTC GGA AAT 2116 lie Gin Val Thr Cys Arg Glu lie Pro lie Lys Ser Val Val Gly Asn

685 690 695

ACC ACG ATG ATG GAA CAG GTT TTG GAA TTT CTC AGT GAA GAA GAA GAA 2164 Thr Thr Met Met Glu Gin Val Leu Glu Phe Leu Ser Glu Glu Glu Glu

700 705 710

AGA GGA ATC ATT GGT GTT TAT GGA CCT GGT GGG GTT GGG AAG ACA ACG 2212 Arg Gly lie lie Gly Val Tyr Gly Pro Gly Gly Val Gly Lys Thr Thr

715 720 725

TTA ATG CAG AGC ATT AAC AAC GAG CTG ATC ACA AAA GGA CAT CAG TAT 2260 Leu Met Gin Ser lie Asn Asn Glu Leu lie Thr Lys Gly His Gin Tyr

730 735 740

GAT GTA CTG ATT TGG GTT CAA ATG TCC AGA GAA TTC GGC GAG TGT ACA 2308 Asp Val Leulie Trp Val Gin Met Ser Arg Glu Phe Gly Glu Cys Thr

745 750 755 760

ATT CAG CAA GCC GTT GGA GCA CGG TTG GGT TTA TCT TGG GAC GAG AAG 2356 lie Gin Gin Ala Val Gly Ala Arg Leu Gly Leu Ser Trp Asp Glu Lys

765 770 775

GAG ACC GGC GAA AAC AGA GCT TTG AAG ATA TAC AGA GCT TTG AGA CAG 2404 Glu Thr Gly Glu Asn Arg Ala Leu Lys lie Tyr Arg Ala Leu Arg Gin

780 785 790

AAA CGT TTC TTG TTG TTG CTA GAT GAT GTC TGG GAA GAG ATA GAC TTG 2452 Lys Arg Phe Leu Leu Leu Leu Asp Asp Val Trp Glu Glu lie Asp Leu 795 800 805

GAG AAA ACT GGA GTT CCT CGA CCT GAC AGG GAA AAC AAA TGC AAG GTG 2500 Glu Lys Thr Gly Val Pro Arg Pro Asp Arg Glu Asn Lys Cys Lys Val

810 815 820

ATG TTC ACG ACA CGG TCT ATA GCA TTA TGC AAC AAT ATG GGT GCG GAA 2548 Met Phe Thr Thr Arg Ser lie Ala Leu Cys Asn Asn Met Gly Ala Glu

825 830 835 840

TAC AAG TTG AGA GTG GAG TTT CTG GAG AAG AAA CAC GCG TGG GAG CTG 2596 Tyr Lys Leu Arg Val Glu Phe Leu Glu Lys Lys His Ala Trp Glu Leu

845 850 855

TTC TGT AGT AAG GTA TGG AGA AAA GAT CTT TTA GAG TCA TCA TCA ATT 2644 Phe Cys Ser Lys Val Trp Arg Lys Asp Leu Leu Glu Ser Ser Serlie

860 865 870

CGC CGG CTC GCG GAG TGAATTC 2666 Arg Arg Leu Ala Glu

875 SEQ ID NO: 9

Sequence length: 683

Sequence type: amino acid

Topology: linear

Sequence type: protein

Array:

Asn Ala Phe Ser Gly lie He Arg lie Ala Leu Leu Pro Asp Ser Asp

1 5 10 15

Ser Lys His Glu Ala Val Leu Asp Lys Tyr Ser Ser Cys Tyr Pro Val

20 25 30

Ser Gly Lys Ala Val Phe Arg Glu Pro Phe Cys Val Glu Tyr Asn Trp

35 40 45 Glu Lys Lys Asp Ser Gly Asp Leu Leu Leu Leu Ala His Pro Leu His

50 55 60

Val Gin Leu Leu Arg Asn Gly Asp Asn Asp Val Lys lie Leu Glu Asp 65 70 75 80

Leu Lys Tyr Lys Ser lie Asp Gly Asp Leu Val Gly Val Val Gly Asp

85 90 95

Ser Trp Val Leu Lys Thr Asp Pro Leu Phe Val Thr Trp His Ser lie

100 105 110

Lys Gly I le Lys Glu Glu Ser His Asp Glu lie Val Ser Ala Leu Ser

115 120 125

Lys Asp Val Glu Ser Leu Asp Ser Ser Ser lie Thr Thr Thr Glu Ser

130 135 140

Tyr Phe Tyr Gly Lys Leu lie Ala Arg Ala Ala Arg Leu Val Leu lie 145 150 155 160

Ala Glu Glu Leu Asn Tyr Pro Asp Val lie Pro Lys Val Arg Asn Phe

165 170 175

Leu Lys Glu Thr lie Glu Pro Trp Leu Glu Gly Thr Phe Ser Gly Asn

180 185 190

Gly Phe Leu His Asp Glu Lys Trp Gly Gly lie lie Thr Gin Lys Gly

195 200 205

Ser Thr Asp Ala Gly Gly Asp Phe Gly Phe Gly lie Tyr Asn Asp His

210 215 220

His Tyr His Leu Gly Tyr Phe lie Tyr Gly lie Ala Val Leu Thr Lys 225 230 235 240

Leu Asp Pro Ala Trp Gly Arg Lys Tyr Lys Pro Gin Ala Tyr Ser lie

245 250 255

Val Gin Asp Phe Leu Asn Leu Asp Thr Lys Leu Asn Ser Asn Tyr Thr

260 265 270

Arg Leu Arg Cys Phe Asp Pro Tyr Val Leu His Ser Trp Ala Gly Gly 275 280 285

Leu Thr Glu Phe Thr Asp Gly Arg Asn Gin Glu Ser Thr Ser Glu Ala

290 295 300

Val Ser Ala Tyr Tyr Ser Ala Ala Leu Met Gly Leu Ala Tyr Gly Asp 305 310 315 320

Ala Pro Leu Val Ala Leu Gly Ser Thr Leu Thr Ala Leu Glu He Glu

325 330 335

Gly Thr Lys Met Trp Trp His Ala Met Asp Phe lie Ser Ser Leu lie

340 345 350

Val Gly Cys Ala Gin Val Leu Cys Glu Ser Met Asn Met Ala Glu Arg

355 360 365

Arg Gly His Lys Thr Asp Leu Arg Gin Ala lie Thr Asp Leu Glu Thr

370 375 380

Ala lie Gly Asp Leu Lys Ala lie Arg Asp Asp Leu Thr Leu Arg lie 385 390 395 400

Gin Gin Asp Gly Leu Glu Gly Arg Ser Cys Ser Asn Arg Ala Arg Glu

405 410 415

Trp Leu Ser Ala Val Gin Val Thr Glu Thr Lys Thr Ala Leu Leu Leu

420 425 430

Val Arg Phe Arg Arg Arg Glu Gin Arg Thr Arg Met Arg Arg Arg Tyr

435 440 445

Leu Ser Cys Phe Gly Cys Ala Asp Tyr Lys Leu Cys Lys Lys Val Ser

450 455 460

Ala lie Leu Lys Ser lie Gly Glu Leu Arg Glu Arg Ser Glu Ala lie 465 470 475 480

Lys Thr Asp Gly Gly Ser lie Gin Val Thr Cys Arg Glu lie Pro lie

485 490 495

Lys Ser Val Val Gly Asn Thr Thr Met Met Glu Gin Val Leu Glu Phe

500 505 510 Leu Ser Glu Glu Glu Glu Arg Gly lie lie Gly Val Tyr Gly Pro Gly

515 520 525

Gly Val Gly Lys Thr Thr Leu Met Gin Ser lie Asn Asn Glu Leu lie

530 535 540

Thr Lys Gly His Gin Tyr Asp Val Leu lie Trp Val Gin Met Ser Arg 545 550 555 560

Glu Phe Gly Glu Cys Thr lie Gin Gin Ala Val Gly Ala Arg Leu Gly

565 570 575

Leu Ser Trp Asp Glu Lys Glu Thr Gly Glu Asn Arg Ala Leu Lys lie

580 585 590

Tyr Arg Ala Leu Arg Gin Lys Arg Phe Leu Leu Leu Leu Asp Asp Val

595 600 605

Trp Glu Glu lie Asp Leu Glu Lys Thr Gly Val Pro Arg Pro Asp Arg

610 615 620

Glu Asn Lys Cys Lys Val Met Phe Thr Thr Arg Ser lie Ala Leu Cys 625 630 635 640

Asn Asn Met Gly Ala Glu Tyr Lys Leu Arg Val Glu Phe Leu Glu Lys

645 650 655

Lys His Ala Trp Glu Leu Phe Cys Ser Lys Val Trp Arg Lys Asp Leu

660 665 670

Leu Glu Ser Ser Ser lie Arg Arg Leu Ala Glu

675 680 SEQ ID NO: 10

Sequence length: 2117

Sequence type: nucleic acid

Number of chains: double strand

Topology: linear

Sequence type: cDNA to mRNA Array features

Characteristic symbol: CDS

Location: 62..2110

Array:

GCGGCCGCCT TCTTTCCTCA ACCTTCTTTC TTCTTATATA TTCGAACAAT TCGATATCAT 60 G AAT GCA TTT TCT GGC ATA ATC CGG ATA GCT TTG TTG CCG GAT TCG 106 Asn Ala Phe Ser Gly lie lie Arg lie Ala Leu Leu Pro Asp Ser

1 5 10 15

GAT TCG AAA CAC GAG GCT GTT CTT GAC AAG TAT AGT TCT TGT TAC CCC 154 Asp Ser Lys His Glu Ala Val Leu Asp Lys Tyr Ser Ser Cys Tyr Pro

20 25 30

GTG TCA GGT AAA GCT GTG TTC AGA GAA CCT TTC TGT GTG GAA TAT AAC 202 Val Ser Gly Lys Ala Val Phe Arg Glu Pro Phe Cys Val Glu Tyr Asn

35 40 45

TGG GAG AAG AAA GAT TCA GGG GAT TTG CTA CTC TTG GCT CAC CCT CTC 250 Trp Glu Lys Lys Asp Ser Gly Asp Leu Leu Leu Leu Ala His Pro Leu

50 55 60

CAT GTT CAG CTT CTT CGT AAT GGA GAC AAT GAT GTC AAA ATT CTT GAA 298 His Val Gin Leu Leu Arg Asn Gly Asp Asn Asp Val Lys lie Leu Glu

65 70 75

GAT TTA AAG TAT AAA AGC ATT GAT GGG GAT CTT GTT GGT GTT GTC GGG 346 Asp Leu Lys Tyr Lys Ser lie Asp Gly Asp Leu Val Gly Val Val Gly

80 85 90 95

GAT TCA TGG GTT TTG AAA ACA GAT CCT TTG TTT GTA ACA TGG CAT TCA 394 Asp Ser Trp Val Leu Lys Thr Asp Pro Leu Phe Val Thr Trp His Ser

100 105 110

ATC AAG GGA ATC AAA GAA GAA TCC CAT GAT GAG ATT GTC TCA GCC CTT 442 lie Lys Gly lie Lys Glu Glu Ser His Asp Glu lie Val Ser Ala Leu

115 120 125 TCT AAA GAT GTT GAG AGC CTA GAT TCA TCA TCA ATA ACT ACA ACA GAG 490 Ser Lys Asp Val Glu Ser Leu Asp Ser Ser Ser lie Thr Thr Thr Glu

130 135 140

TCA TAT TTT TAT GGG AAG TTG ATT GCA AGG GCT GCA AGG TTG GTA TTG 538 Ser Tyr Phe Tyr Gly Lys Leu I le Ala Arg Ala Ala Arg Leu Val Leu

145 150 155

ATT GCT GAG GAG TTG AAC TAC CCT GAT GTG ATT CCA AAG GTT AGG AAT 586 lie Ala Glu Glu Glu Leu Asn Tyr Pro Asp Val lie Pro Lys Val Arg Asn

160 165 170 175

TTT TTG AAA GAA ACC ATT GAG CCA TGG TTG GAG GGA ACT TTT AGT GGG 634 Phe Leu Lys Glu Thr lie Glu Pro Trp Leu Glu Gly Thr Phe Ser Gly

180 185 190

AAT GGA TTC CTA CAT GAT GAA AAA TGG GGT GGC ATT ATT ACC CAA AAG 682 Asn Gly Phe Leu His Asp Glu Lys Trp Gly Gly lie He Thr Gin Lys

195 200 205

GGG TCC ACT GAT GCT GGT GGT GAT TTT GGA TTT GGA ATT TAC AAT GAT 730 Gly Ser Thr Asp Ala Gly Gly Asp Phe Gly Phe Glylie Tyr Asn Asp

210 215 220

CAC CAC TAT CAT TTG GGG TAC TTC ATT TAT GGA ATT GCG GTG CTC ACT 778 His His Tyr His Leu Gly Tyr Phe lie Tyr Gly I le Ala Val Leu Thr

225 230 235

AAG CTT GAT CCA GCA TGG GGT AGG AAG TAC AAG CCT CAA GCC TAT TCA 826 Lys Leu Asp Pro Ala Trp Gly Arg Lys Tyr Lys Pro Gin Ala Tyr Ser

240 245 250 255

ATA GTG CAA GAC TTC TTG AAC TTG GAC ACA AAA TTA AAC TCC AAT TAC 874 I le Val Gin Asp Phe Leu Asn Leu Asp Thr Lys Leu Asn Ser Asn Tyr

260 265 270

ACA CGT TTG AGG TGT TTT GAC CCT TAT GTG CTT CAC TCT TGG GCT GGA 922 Thr Arg Leu Arg Cys Phe Asp Pro Tyr Val Leu His Ser Trp Ala Gly 275 280 285

GGG TTA ACT GAG TTC ACA GAT GGA AGG AAT CAA GAG AGC ACA AGT GAG 970 Gly Leu Thr Glu Phe Thr Asp Gly Arg Asn Gin Glu Ser Thr Ser Glu

290 295 300

GCT GTG AGT GCA TAT TAT TCT GCT GCT TTG ATG GGA TTA GCA TAT GGT 1018 Ala Val Ser Ala Tyr Tyr Ser Ala Ala Leu Met Gly Leu Ala Tyr Gly

305 310 315

GAT GCA CCT CTT GTT GCA CTT GGA TCA ACA CTC ACA GCA TTG GAA ATT 1066 Asp Ala Pro Leu Val Ala Leu Gly Ser Thr Leu Thr Ala Leu Glulie

320 325 330 335

GAA GGG ACT AAA ATG TGG TGG CAT GCG ATG GAT TTC ATC TCA TCT CTT 1114 Glu Gly Thr Lys Met Trp Trp His Ala Met Asp Phelie Ser Ser Leu

340 345 350

ATC GTT GGC TGT GCT CAG GTG TTG TGT GAA TCT ATG AAT ATG GCG GAG 1162 lie Val Gly Cys Ala Gin Val Leu Cys Glu Ser Met Asn Met Ala Glu

355 360 365

AGA AGA GGA CAT AAG ACT GAT CTT AGA CAA GCC ATC ACT GAT CTT GAA 1210 Arg Arg Gly His Lys Thr Asp Leu Arg Gin Ala lie Thr Asp Leu Glu

370 375 380

ACA GCC ATC GGT GAC TTG AAG GCC ATA CGT GAT GAC CTG ACT TTA CGG 1258 Thr Ala lie Gly Asp Leu Lys Ala lie Arg Asp Asp Leu Thr Leu Arg

385 390 395

ATC CAA CAA GAC GGT CTA GAG GGA CGA AGC TGC TCA AAT CGT GCC AGA 1306 He Gin Gin Asp Gly Leu Glu Gly Arg Ser Cys Ser Asn Arg Ala Arg

400 405 410 415

GAG TGG CTT AGT GCG GTG CAA GTA ACG GAG ACT AAA ACA GCC CTA CTT 1354 Glu Trp Leu Ser Ala Val Gin Val Thr Glu Thr Lys Thr Ala Leu Leu

420 425 430

TTA GTG AGG TTT AGG CGT CGG GAA CAG AGG ACG CGA ATG AGG AGG AGA 1402 Leu Val Arg Phe Arg Arg Arg Glu Gin Arg Thr Arg Met Arg Arg Arg

435 440 445

TAC CTC AGT TGT TTC GGT TGT GCC GAC TAC AAA CTG TGC AAG AAG GTT 1450 Tyr Leu Ser Cys Phe Gly Cys Ala Asp Tyr Lys Leu Cys Lys Lys Val

450 455 460

TCT GCC ATA TTG AAG AGC ATT GGT GAG CTG AGA GAA CGC TCT GAA GCT 1498 Ser Ala lie Leu Lys Ser lie Gly Glu Leu Arg Glu Arg Ser Glu Ala

465 470 475

ATC AAA ACA GAT GGC GGG TCA ATT CAA GTA ACT TGT AGA GAG ATA CCC 1546 He Lys Thr Asp Gly Gly Serlie Gin Val Thr Cys Arg Glu He Pro

480 485 490 495

ATC AAG TCC GTT GTC GGA AAT ACC ACG ATG ATG GAA CAG GTT TTG GAA 1594 lie Lys Ser Val Val Gly Asn Thr Thr Met Met Glu Gin Val Leu Glu

500 505 510

TTT CTC AGT GAA GAA GAA GAA AGA GGA ATC ATT GGT GTT TAT GGA CCT 1642 Phe Leu Ser Glu Glu Glu Glu Arg Gly lie lie Gly Val Tyr Gly Pro

515 520 525

GGT GGG GTT GGG AAG ACA ACG TTA ATG CAG AGC ATT AAC AAC GAG CTG 1690 Gly Gly Val Gly Lys Thr Thr Leu Met Gin Ser lie Asn Asn Glu Leu

530 535 540

ATC ACA AAA GGA CAT CAG TAT GAT GTA CTG ATT TGG GTT CAA ATG TCC 1738 lie Thr Lys Gly His Gin Tyr Asp Val Leu lie Trp Val Gin Met Ser

545 550 555

AGA GAA TTC GGC GAG TGT ACA ATT CAG CAA GCC GTT GGA GCA CGG TTG 1786 Arg Glu Phe Gly Glu Cys Thrlie Gin Gin Ala Val Gly Ala Arg Leu

560 565 570 575

GGT TTA TCT TGG GAC GAG AAG GAG ACC GGC GAA AAC AGA GCT TTG AAG 1834 Gly Leu Ser Trp Asp Glu Lys Glu Thr Gly Glu Asn Arg Ala Leu Lys

580 585 590 ATA TAC AGA GCT TTG AGA CAG AAA CGT TTC TTG TTG TTG CTA GAT GAT 1882 l ie Tyr Arg Ala Leu Arg Gin Lys Arg Phe Leu Leu Leu Leu Asp Asp

595 600 605

GTC TGG GAA GAG ATA GAC TTG GAG AAA ACT GGA GTT CCT CGA CCT GAC 1930 Val Trp Glu Glu I le Asp Leu Glu Lys Thr Gly Val Pro Arg Pro Asp

610 615 620

AGG GAA AAC AAA TGC AAG GTG ATG TTC ACG ACA CGG TCT ATA GCA TTA 1978 Arg Glu Asn Lys Cys Lys Val Met Phe Thr Thr Arg Ser I le Ala Leu

625 630 635

TGC AAC AAT ATG GGT GCG GAA TAC AAG TTG AGA GTG GAG TTT CTG GAG 2026 Cys Asn Asn Met Gly Ala Glu Tyr Lys Leu Arg Val Glu Phe Leu Glu

640 645 650 655

AAG AAA CAC GCG TGG GAG CTG TTC TGT AGT AAG GTA TGG AGA AAA GAT 2074 Lys Lys His Ala Trp Glu Leu Phe Cys Ser Lys Val Trp Arg Lys Asp

660 665 670

CTT TTA GAG TCA TCA TCA ATT CGC CGG CTC GCG GAG TGAATTC 2117 Leu Leu Glu Ser Ser Ser I le Arg Arg Leu Ala Glu

675 680 SEQ ID NO: 11

Sequence length: 347

Sequence type: amino acid

Topology: linear

Sequence type: protein

Array:

Met Ala Lys Tyr His Ser Ser Gly Lys Ser Ser Ser Met Thr Ala lie

1 5 10 15

Ala Phe Leu Phe lie Leu Leu 1 le Thr Tyr Thr Gly Thr Thr Asp Ala

20 25 30 in Ser Gly Val Cys Tyr Gly Arg Leu Gly Asn Asn Leu Pro Thr Pro

35 40 45

in Glu Val Val Ala Leu Tyr Asn Gin Ala Asn lie Arg Arg Met Arg

50 55 60

lie Tyr Gly Pro Ser Pro Glu Val Leu Glu Ala Leu Arg Gly Ser Asn 65 70 75 80 lie Glu Leu Leu Leu Asp I le Pro Asn Asp Asn Leu Arg Asn Leu Ala

85 90 95

Ser Ser Gin Asp Asn Ala Asn Lys Trp Val Gin Asp Asn lie Lys Asn

100 105 110

Tyr Ala Asn Asn Val Arg Phe Arg Tyr Val Ser Val Gly Asn Glu Val

115 120 125

Lys Pro Glu His Ser Phe Ala Gin Phe Leu Val Pro Ala Leu Glu Asn

130 135 140

lie Gin Arg Ala lie Ser Asn Ala Gly Leu Gly Asn Gin Val Lys Val 145 150 155 160

Ser Thr Ala lie Asp Thr Gly Ala Leu Ala Glu Ser Phe Pro Pro Ser

165 170 175

Lys Gly Ser Phe Lys Ser Asp Tyr Arg Gly Ala Tyr Leu Asp Gly Val

180 185 190 lie Arg Phe Leu Val Asn Asn Asn Ala Pro Leu Met Val Asn Val Tyr

195 200 205

Ser Tyr Phe Ala Tyr Thr Ala Asn Pro Lys Asp lie Ser Leu Asp Tyr

210 215 220

Ala Leu Phe Arg Ser Pro Ser Val Val Val Gin Asp Gly Ser Leu Gly 225 230 235 240

Tyr Arg Asn Leu Phe Asp Ala Ser Val Asp Ala Val Tyr Ala Ala Leu

245 250 255

Glu Lys Ala Gly Gly Gly Ser Leu Asn lie Val Val Ser Glu Ser Gly 260 265 270

Trp Pro Ser Ser Gly Gly Thr Ala Thr Ser Leu Asp Asn Ala Arg Thr

275 280 285

Tyr Asn Thr Asn Leu Val Arg Asn Val Lys Gin Gly Thr Pro Lys Arg

290 295 300

Pro Gly Ala Pro Leu Glu Thr Tyr Val Phe Ala Met Phe Asp Glu Asn

305 310 315 320

Gin Lys Gin Pro Glu Phe Glu Lys Phe Trp Gly Leu Phe Ser Pro lie

325 330 335

Thr Lys Gin Pro Lys Tyr Ser lie Asn Phe Asn

340 345 SEQ ID NO: 12

Array length: 1044

Sequence type: nucleic acid

Number of chains: double strand

Topology: linear

Sequence type: cDNA to mRNA

Array:

ATG GCT AAG TAT CAT TCA AGT GGG AAA AGC TCT TCC ATG ACT GCT ATA 48 Met Ala Lys Tyr His Ser Ser Gly Lys Ser Ser Ser Met Thr Ala lie

1 5 10 15

GCC TTC CTG TTT ATC CTT CTA ATC ACT TAT ACA GGC ACA ACA GAT GCA 96 Ala Phe Leu Phe lie Leu Leu lie Thr Tyr Thr Gly Thr Thr Asp Ala

20 25 30

CAA TCC GGG GTA TGT TAT GGA AGA CTT GGC AAC AAC TTA CCA ACC CCT 144 Gin Ser Gly Val Cys Tyr Gly Arg Leu Gly Asn Asn Leu Pro Thr Pro

35 40 45

CAA GAA GTT GTG GCC CTC TAC AAT CAA GCC AAC ATT CGC AGG ATG CGA 192 Gin Glu Val Val Ala Leu Tyr Asn Gin Ala Asn lie Arg Arg Met Arg

50 55 60

ATC TAC GGT CCA AGC CCA GAA GTC CTC GAA GCA CTA AGA GGT TCC AAC 240 lie Tyr Gly Pro Ser Pro Glu Val Leu Glu Ala Leu Arg Gly Ser Asn

65 70 75 80

ATT GAG CTT TTG CTA GAC ATT CCA AAT GAC AAC CTC AGA AAC CTA GCA 288 lie Glu Leu Leu Leu Asp lie Pro Asn Asp Asn Leu Arg Asn Leu Ala

85 90 95

TCT AGC CAA GAC AAT GCA AAC AAA TGG GTG CAA GAC AAC ATC AAA AAC 336 Ser Ser Gin Asp Asn Ala Asn Lys Trp Val Gin Asp Asn lie Lys Asn

100 105 110

TAT GCC AAC AAT GTC AGA TTC AGA TAC GTT TCA GTG GGA AAT GAA GTG 384 Tyr Ala Asn Asn Val Arg Phe Arg Tyr Val Ser Val Gly Asn Glu Val

115 120 125

AAA CCC GAA CAC TCA TTT GCA CAA TTT CTA GTG CCT GCA TTG GAA AAC 432 Lys Pro Glu His Ser Phe Ala Gin Phe Leu Val Pro Ala Leu Glu Asn

130 135 140

ATT CAG AGG GCC ATT TCT AAT GCT GGC CTT GGA AAC CAA GTA AAA GTT 480 lie Gin Arg Ala lie Ser Asn Ala Gly Leu Gly Asn Gin Val Lys Val 145 150 155 160

TCC ACT GCC ATT GAT ACT GGT GCC TTG GCA GAA TCA TTC CCA CCA TCA 528 Ser Thr Ala lie Asp Thr Gly Ala Leu Ala Glu Ser Phe Pro Pro Ser

-165 170 175

AAG GGT TCC TTC AAA TCT GAT TAT AGA GGA GCA TAT CTT GAT GGT GTC 576 Lys Gly Ser Phe Lys Ser Asp Tyr Arg Gly Ala Tyr Leu Asp Gly Val

180 185 190

ATC AGA TTT CTA GTG AAC AAT AAT GCC CCA TTA ATG GTT AAT GTG TAC 624 lie Arg Phe Leu Val Asn Asn Asn Ala Pro Leu Met Val Asn Val Tyr

195 200 205 OK

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ΘΙ Ι OJJ J8S Slid nsi Aio dj 丄 9 sAq nio nio OJJ UIO SAI uio 8001 VIV 100 131 III DID DOO DDI III VVV VVO III OVO VOD OVO OVV OVD

ozz 9ΐε οτε 9οε usv nio dsy aqj 13 ^ ^ iv sqj Ι¾Λ 丄 `` io naq OJJ BIV IO OJJ 096 IVV VVO IVD 111 DIV 330 III DID IV 丄 IDV VVO 113 0D3 VOO 100 丄 33

00S S6Z 062

3JV s OJJ jqi u] 9 sAq eight usy S-iy Ι¾Λ "91 usy J¾ usy αΛχ

Ζΐ6 90V VVV 133 3DV VOD VVO OVV DID IVV DOD 110 Oil OVV VOV OVV 3VI

SSZ 08Z 2LZ

JiU SJV BIV usv dsy "91 ¾IV Jqi ^ IO ^ IO J3S J9S OJJ dj 丄 98 10V VOV VOO IVV IVO 113 V31 IDV VOD I3V VOD IDD 131 131 103 001

OLZ 9Z 09Z

^ ID niO ^J9 S Ι ^ Λ Ι¾Λ 811 usy nai Jas Λιο λιο Ajo ¾ιγ sAq nio

918 VOD IDV OVO 131 DID IID VIV OVV Oil VD1 ODD VDO VDO VOD VVV OVO

Z 0 Z Z

naq ¾iy V

89 Oil V30 100 IVl 110 100 IVO IID DDI 100 IVO 111 013 OVV IDD DVI

O Z 2Z 0 Z 9ZZ

^ IO "9Ί J8S ^ IO dsv uio Ι ^ Λ Ι ^ Λ Ι¾Λ OJJ jas §JV 3Ud naq ¾iv OZ 100 113 V31 IDO IVO WD 010 VIO DID 031 130 131 OOV HI IID VOO

OZZ 1Z 01Z 丄丄 dsy neq d \ \ dsy sAi OJJ usy ¾iv Jqi 丄 ¾iv aqd 丄 Z / .9 IVl OVD 113 10V IIV DVD OVV 133 OVV VOO I3V 3V1 IDD Oil DVI 丄 :) 丄

6 Z0 / 86df / 13d S908S / 86 OM Array length: 331

Sequence type: amino acid

Topology: linear

Sequence type: protein

Array:

Met ¾er Ala Leu Leu Leu Leu Leu Gly Val Leu Ser Ser Thr Gly Val

1 5 10 15

Leu Leu Thr Gly Val Glu Ser Val Gly Val Cys Tyr Gly Gly Asn Gly

20 25 30

Asn Asn Leu Pro Thr Lys Gin Ala Val Val Asn Leu Tyr Lys Ser Asn

35 40 45

Gly lie Gly Lys lie Arg Leu Tyr Tyr Pro Asp Glu Gly Ala Leu Gin

50 55 60

Ala Leu Arg Gly Ser Asn lie Glu Val lie Leu Ala Val Pro Asn Asp 65 70 75 80

Gin Leu Gin Ser Val Ser Asn Asn Gly Gly Ala Thr Asn Trp Val Asn

85 90 95

Lys Tyr Val Lys Pro Tyr Ala Gly Asn Val Lys Leu Lys Tyr lie Ala

100 105 110

Val Gly Asn Glu Val His Pro Gly Asp Ala Leu Ala Gly Ser Val Leu

115 120 125

Pro Ala Leu Gin Ser I le Gin Asn Ala lie Ser Ala Ala Asn Leu Gin

130 135 140

Arg Gin lie Lys Val Ser Thr Ala lie Asp Thr Thr Leu Leu Gly Asn 145 150 155 160

Ser Tyr Pro Pro Lys Asp Gly Val Phe Ser Asn Ser Ala Ser Ser Tyr

165 170 175 lie Thr Pro I le lie Asn Phe Leu Ala Lys Asn Gly Ala Pro Leu Leu

180 185 190 Ala Asn Val Tyr Pro Tyr Phe Ala Tyr Val Asn Asn Gin Gin Asn lie

195 200 205

Gly Leu Asp Tyr Ala Leu Phe Thr Lys Gin Gly Asn Asn Glu Val Gly

210 215 220

Tyr Gin Asn Leu Phe Asp Ala Leu Val Asp Ser Leu Tyr Ala Ala Leu

225 230 235 240

Glu Lys Val Gly Ala Ser Asn Val Lys Val Val Val Ser Glu Ser Gly

245 250 255

Trp Pro Ser Gin Gly Gly Val Gly Ala Thr Val Gin Asn Ala Gly Thr

260 265 270

Tyr Tyr Arg Asn Leu lie Lys His Val Lys Gly Gly Thr Pro Lys Arg

275 280 285

Pro Asn Gly Pro lie Glu Thr Tyr Leu Phe Ala Met Phe Asp Glu Asn

290 295 300

Gin Lys Gly Gly Ala Glu Thr Glu Lys His Phe Gly Leu Phe Arg Pro

305 310 315 320

Asp Lys Ser Pro Lys Tyr Gin Leu Ser Phe Asn

325 330 SEQ ID NO: 14

Sequence length: 993

Sequence type: nucleic acid

Number of chains: double strand

Topology: linear

Sequence type: cDNA to mRNA

Array:

ATG TCT GCC TTA TTG CTG CTT CTT GGA GTA TTA TCT TCC ACT GGA GTA 48 Met Ser Ala Leu Leu Leu Leu Leu Gly Val Leu Ser Ser Thr Gly Val

1 5 10 15 CTG CTT ACT GGG GTA GAA TCT GTG GGT GTG TGT TAT GGA GGA AAT GGA 96 Leu Leu Thr Gly Val Glu Ser Val Gly Val Cys Tyr Gly Gly Asn Gly

20 25 30

AAC AAT CTA CCA ACA AAG CAA GCA GTG GTG AAT CTC TAC AAA TCA AAC 144 Asn Asn Leu Pro Thr Lys Gin Ala Val Val Asn Leu Tyr Lys Ser Asn

35 40 45

GGA ATT GGC AAA ATC CGT TTA TAC TAT CCA GAT GAA GGT GCC CTT CAA 192 Gly lie Gly Lys lie Arg Leu Tyr Tyr Pro Asp Glu Gly Ala Leu Gin

50 55 60

GCC CTC AGA GGT TCA AAC ATA GAA GTG ATA CTT GCT GTT CCT AAT GAT 240 Ala Leu Arg Gly Ser Asn lie Glu Val lie Leu Ala Val Pro Asn Asp

65 70 75 80

CAA CTT CAA TCT GTC TCC AAC AAT GGA GGT GCA ACA AAT TGG GTC AAC 288 Gin Leu Gin Ser Val Ser Asn Asn Gly Gly Ala Thr Asn Trp Val Asn

85 90 95

AAG TAC GTG AAA CCC TAT GCA GGA AAC GTG AAA TTG AAG TAC ATT GCA 336 Lys Tyr Val Lys Pro Tyr Ala Gly Asn Val Lys Leu Lys Tyr lie Ala

100 105 110

GTT GGC AAC GAA GTT CAC CCT GGT GAT GCT CTA GCA GGC TCA GTT CTT 384 Val Gly Asn Glu Val His Pro Gly Asp Ala Leu Ala Gly Ser Val Leu

115 120 125

CCA GCA CTT CAA AGC ATT CAG AAC GCA ATT TCT GCA GCA AAT TTG CAA 432 Pro Ala Leu Gin Ser lie Gin Asn Ala lie Ser Ala Ala Asn Leu Gin

130 135 140

CGC CAA ATC AAA GTC TCC ACA GCA ATA GAC ACC ACT CTA CTG GGC AAC 480 Arg Gin ly Lys Val Ser Thr Ala lie Asp Thr Thr Leu Leu Gly Asn

145 150 155 160

TCT TAC CCA CCA AAA GAT GGC GTT TTC AGC AAC AGT GCA AGT TCA TAC 528 Ser Tyr Pro Pro Lys Asp Gly Val Phe Ser Asn Ser Ala Ser Ser Tyr C66 IVV 311 10V DID VV3 OVl VVV VDD VOl VVV IVO

^{0Jd SJV 94d ^ 10 9 ¾1 S} IH niO J ¾ n ID ^ IV ^ IO ^ ID sAq u \ ^) 096 133 ODV Oil DID IDO 111 OVO VVV OVO I3V WO VDO 100 IDO OVV OVO

OOC S6Z 062 usv ⁿ I3 dsy 94d

usy OJJ 216 OVV WD IVO 111 DIV ODD III DID DVl I3V OVO VIV 330 VDO IVV 丄 3 :)

Z 02Z LZ

SJV sA ojj jqi 3 io sA Ι¾Λ S IH sAq 9i! Na usy §jy J ^ l 丄 98 ODV OVV VOO DDV ODD IDD OVV IID IVO VVV OIV DII IVV ODV OVl IV 丄

OLZ 992 09Z

丄 9 ¾iv usv uio Ι¾Λ jqi EIV AIO Ι¾Λ ^ IO uio s OJJ dj 丄

918 ODV VOO VOO OVV VV3 IID IOV D30 VDD 110 VDD 100 VV3 VOl VOO 001

Z 0 Z Z

lO J9S "ID Ι Ι ^ Λ Ι ^ Λ ^s Ί Ι¾Λ usv ¾IV ^ IO 1¾Λ sAq

89 ODD IOV OVO IDl 010 110 IID OVV 010 IVV VOl VDO VDO 010 VVV OVO

OZ 9 £ Z OZZ ZZ na B v ¾IV丄^{^{ja§ dsy ΐ¾Λ n9 l ^ IV ds}} V 9Md na l usy u {)丄

0 113 IDO VOO DVl 013 IDl IVO DID Oil V3D IVD III DID DVV VV3 OVl

OZZ ζ \ Ζ ΟΐΖ IO Ι ^ Λ ^ io usy usv Λιο U \ D sA J¾ sqd nsi BJVΐλ 丄 dsy na Λιο

ZL9 DDO 110 WD OVV OVV 300 VVD VVV 33V 111 Oil 330 IVl IVO 110 IDO

03 OOZ S61

9] I usv uio uio usv usy Ι ^ Λ ¾IV ^ ά "^ 丄 OJJ JA 丄 Ι¾Λ usy ¾iv

^ 9 11V OVV WD VV3 IVV DVV 110 OVl 330 311 OVl IDD OVl DID OVV VOO

061 S8I 081

na ^] Tisi OJJ BJV 19 usy sA- Bfy na aqj usy ei I d \ \ OJJ 丄

9 S IID 113 VOO DDO IDO OVV VVV DDO Vll 111 DVV VIV OIV VDD IDV VIV

SL1 0Z.1 S91

6 I0 / 86Jf / 13d S908S / 86 OM sp Lys Ser Pro Lys Tyr Gin Leu Ser Phe Asn

325 330 SEQ ID NO: 15

Array length: 34

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

CTAGAGGATC CGGTACCCCC GGGGTCGACG AGCT 34 SEQ ID NO: 16

Array length: 26

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

CGTCGACCCC GGGGGTACCG GATCCT 26 SEQ ID NO: 17

Array length: 4

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

CTAGACTTCT TTCCTCAACC TTCTTTCTTC TTATATATTC GAAC 44 SEQ ID NO: 18

Array length: 44

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

GATCGTTCGA ATATATAAGA AGAAAGAAGG TTGAGGAAAG AAGT 44 SEQ ID NO: 19

Array length: 30

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

TGGCATGCGA TGGATTTCAT CTCATCTCTT 30 SEQ ID NO: 20

Array length: 27

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

GGGAATTCAC TCCGCGAGCC GGCGAAT 27 SEQ ID NO: 21 Array length: 35

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

ACAAGTAAAA GAAAGAGCGA GAAATCATCG AAATG 35 SEQ ID NO: 22

Array length: 24

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

AGCCATGGCT CCTCCTAAAG TGAT 24 SEQ ID NO: 23

Array length: 21

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

GGCCGCGATA TCATGAATGC A 21 SEQ ID NO: 24

Array length:

Sequence type: nucleic acid Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

TTCATGATAT CGC 13 SEQ ID NO: 25

Array length: 45

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

GGCCGCCTTC TTTCCTCAAC CTTCTTTCTT CTTATATATT CGAAC 45 SEQ ID NO: 26

Array length: 45

Sequence type: nucleic acid

Number of chains: single strand

Topology: linear

Sequence type: Other nucleic acids (synthetic DNA)

Array:

AATTGTTCGA ATATATAAGA AGAAAGAAGG TTGAGGAAAG AAGGC 45 SEQ ID NO: 27

Array length: 204

Sequence type: amino acid

Topology: linear

Array type: Array:

Glu Tyr Asn Trp Glu Lys Lys Asp Ser Gly Asp Leu Leu Leu Leu Ala

1 5 10 15

His Pro Leu His Val Gin Leu Leu Arg Asn Gly Asp Asn Asp Val Lys

20 25 30 lie Leu Glu Asp Leu Lys Tyr Lys Ser I le Asp Gly Asp Leu Val Gly

35 40 45

Val Val Gly Asp Ser Trp Val Leu Lys Thr Asp Pro Leu Phe Val Thr

50 55 60

Trp His Ser lie Lys Gly lie Lys Glu Glu Ser His Asp Glu lie Val 65 70 75 80

Ser Ala Leu Ser Lys Asp Val Glu Ser Leu Asp Ser Ser Ser lie Thr

85 90 95

Thr Thr Glu Ser Tyr Phe Tyr Gly Lys Leu lie Ala Arg Ala Ala Arg

100 105 110

Leu Val Leu lie Ala Glu Glu Leu Asn Tyr Pro Asp Val lie Pro Lys

115 120 125

Val Arg Asn Phe Leu Lys Glu Thr lie Glu Pro Trp Leu Glu Gly Thr

130 135 140

Phe Ser Gly Asn Gly Phe Leu His Asp Glu Lys Trp Gly Gly lie I le 145 150 155 160

Thr Gin Lys Gly Ser Thr Asp Ala Gly Gly Asp Phe Gly Phe Gly He

-165 170 175

Tyr Asn Asp His His Tyr His Leu Gly Tyr Phe I le Tyr Gly lie Ala

180 185 190

Val Leu Thr Lys Leu Asp Pro Ala Trp Gly Arg Lys

195 200 SEQ ID NO: 28 Sequence length: 612

Sequence type: nucleic acid

Number of chains: double strand

Topology: linear

Sequence type: cDNA to mRNA

Array:

GAA TAT AAC TGG GAG AAG AAA GAT TCA GGG GAT TTG CTA CTC TTG GCT 48 Glu Tyr Asn Trp Glu Lys Lys Asp Ser Gly Asp Leu Leu Leu Leu Ala

1 5 10 15

CAC CCT CTC CAT GTT CAG CTT CTT CGT AAT GGA GAC AAT GAT GTC AAA 96 His Pro Leu His Val Gin Leu Leu Arg Asn Gly Asp Asn Asp Val Lys

20 25 30

ATT CTT GAA GAT TTA AAG TAT AAA AGC ATT GAT GGG GAT CTT GTT GGT 144 lie Leu Glu Asp Leu Lys Tyr Lys Ser lie Asp Gly Asp Leu Val Gly

35 40 45

GTT GTC GGG GAT TCA TGG GTT TTG AAA ACA GAT CCT TTG TTT GTA ACA 192 Val Val Gly Asp Ser Trp Val Leu Lys Thr Asp Pro Leu Phe Val Thr

50 55 60

TGG CAT TCA ATC AAG GGA ATC AAA GAA GAA TCC CAT GAT GAG ATT GTC 240 Trp His Ser lie Lys Gly He Lys Glu Glu Ser His Asp Glu lie Val 65 70 75 80

TCA GCC CTT TCT AAA GAT GTT GAG AGC CTA GAT TCA TCA TCA ATA ACT 288 Ser Ala Leu Ser Lys Asp Val Glu Ser Leu Asp Ser Ser Serlie Thr

85 90 95

ACA ACA GAG TCA TAT TTT TAT GGG AAG TTG ATT GCA AGG GCT GCA AGG 336 Thr Thr Glu Ser Tyr Phe Tyr Gly Lys Leu lie Ala Arg Ala Ala Arg

100 105 110

TTG GTA TTG ATT GCT GAG GAG TTG AAC TAC CCT GAT GTG ATT CCA AAG 384 Leu Val Leu lie Ala Glu Glu Leu Asn Tyr Pro Asp Val lie Pro Lys 115 120 125

GTT AGG AAT TTT TTG AAA GAA ACC ATT GAG CCA TGG TTG GAG GGA ACT 432 Val Arg Asn Phe Leu Lys Glu Thr lie Glu Pro Trp Leu Glu Gly Thr

130 135 140

TTT AGT GGG AAT GGA TTC CTA CAT GAT GAA AAA TGG GGT GGC ATT ATT 480 Phe Ser Gly Asn Gly Phe Leu His Asp Glu Lys Trp Gly Gly lie lie

145 150 155 160

ACC CAA AAG GGG TCC ACT GAT GCT GGT GGT GAT TTT GGA TTT GGA ATT 528 Thr Gin Lys .Gly Ser Thr Asp Ala Gly Gly Asp Phe Gly Phe Gly He

165 170 175

TAC AAT GAT CAC CAC TAT CAT TTG GGG TAC TTC ATT TAT GGA ATT GCG 576 Tyr Asn Asp His His Tyr His Leu Gly Tyr Phe lie Tyr Gly lie Ala

180 185 190

GTG CTC ACT AAG CTT GAT CCA GCA TGG GGT AGG AAG 612 Val Leu Thr Lys Leu Asp Pro Ala Trp Gly Arg Lys

195 200 SEQ ID NO: 29

Array length: 1080

Sequence type: nucleic acid

Number of chains: double strand

Topology: linear

Sequence type: cDNA to mRNA

Array:

ATG GAT TTC ATC TCA TCT CTT ATC GTT GGC TGT GCT CAG GTG TTG TGT 48 Met Asp Phe lie Ser Ser Leu lie Val Gly Cys Ala Gin Val Leu Cys

1 5 10 15

GAA TCT ATG AAT ATG GCG GAG AGA AGA GGA CAT AAG ACT GAT CTT AGA 96 Glu Ser Met Asn Met Ala Glu Arg Arg Gly His Lys Thr Asp Leu Arg 20 25 30

CAA GCC ATC ACT GAT CTT GAA ACA GCC ATC GGT GAC TTG AAG GCC ATA 144 Gin Ala lie Thr Asp Leu Glu Thr Ala I le Gly Asp Leu Lys Ala lie

35 40 45

CGT GAT GAC CTG ACT TTA CGG ATC CAA CAA GAC GGT CTA GAG GGA CGA 192 Arg Asp Asp Leu Thr Leu Arg lie Gin Gin Asp Gly Leu Glu Gly Arg

50 55 60

AGC TGC TCA AAT CGT GCC AGA GAG TGG CTT AGT GCG GTG CAA GTA ACG 240 Ser Cys Ser Asn Arg Ala Arg Glu Trp Leu Ser Ala Val Gin Val Thr

65 70 75 80

GAG ACT AAA ACA GCC CTA CTT TTA GTG AGG TTT AGG CGT CGG GAA CAG 288 Glu Thr Lys Thr Ala Leu Leu Leu Val Arg Phe Arg Arg Arg Glu Gin

85 90 95

AGG ACG CGA ATG AGG AGG AGA TAC CTC AGT TGT TTC GGT TGT GCC GAC 336 Arg Thr Arg Met Arg Arg Arg Tyr Leu Ser Cys Phe Gly Cys Ala Asp

100 105 110

TAC AAA CTG TGC AAG AAG GTT TCT GCC ATA TTG AAG AGC ATT GGT GAG 384 Tyr Lys Leu Cys Lys Lys Val Ser Ala I le Leu Lys Serlie Gly Glu

115 120 125

CTG AGA GAA CGC TCT GAA GCT ATC AAA ACA GAT GGC GGG TCA ATT CAA 432 Leu Arg Glu Arg Ser Glu Ala I le Lys Thr Asp Gly Gly Serlie Gin

130 135 140

GTA ACT TGT AGA GAG ATA CCC ATC AAG TCC GTT GTC GGA AAT ACC ACG 480 Val Thr Cys Arg Glu lie Pro lie Lys Ser Val Val Gly Asn Thr Thr

145 150 155 160

ATG ATG GAA CAG GTT TTG GAA TTT CTC AGT GAA GAA GAA GAA AGA GGA 528 Met Met Glu Gin Val Leu Glu Phe Leu Ser Glu Glu Glu Glu Arg Gly

165 170 175

ATC ATT GGT GTT TAT GGA CCT GGT GGG GTT GGG AAG ACA ACG TTA ATG 576 lie lie Gly Val Tyr Gly Pro Gly Gly Val Gly Lys Thr Thr Leu Met

180 185 190

CAG AGC ATT AAC AAC GAG CTG ATC ACA AAA GGA CAT CAG TAT GAT GTA 624 Gin Ser lie Asn Asn Glu Leu lie Thr Lys Gly His Gin Tyr Asp Val

195 200 205

CTG ATT TGG GTT CAA ATG TCC AGA GAA TTC GGC GAG TGT ACA ATT CAG 672 Leu lie Trp Val Gin Met Ser Arg Glu Phe Gly Glu Cys Thr lie Gin

210 215 220

CAA GCC GTT GGA GCA CGG TTG GGT TTA TCT TGG GAC GAG AAG GAG ACC 720 Gin Ala Val Gly Ala Arg Leu Gly Leu Ser Trp Asp Glu Lys Glu Thr

225 230 235 240

GGC GAA AAC AGA GCT TTG AAG ATA TAC AGA GCT TTG AGA CAG AAA CGT 768 Gly Glu Asn Arg Ala Leu Lys lie Tyr Arg Ala Leu Arg Gin Lys Arg

245 250 255

TTC TTG TTG TTG CTA GAT GAT GTC TGG GAA GAG ATA GAC TTG GAG AAA 816 Phe Leu Leu Leu Leu Asp Asp Val Trp Glu Glu lie Asp Leu Glu Lys

260 265 270

ACT GGA GTT CCT CGA CCT GAC AGG GAA AAC AAA TGC AAG GTG ATG TTC 864 Thr Gly Val Pro Arg Pro Asp Arg Glu Asn Lys Cys Lys Val Met Phe

275 280 285

ACG ACA CGG TCT ATA GCA TTA TGC AAC AAT ATG GGT GCG GAA TAC AAG 912 Thr Thr Arg Ser lie Ala Leu Cys Asn Asn Met Gly Ala Glu Tyr Lys

290 295 300

TTG AGA GTG GAG TTT CTG GAG AAG AAA CAC GCG TGG GAG CTG TTC TGT 960 Leu Arg Val Glu Phe Leu Glu Lys Lys His Ala Trp Glu Leu Phe Cys

305 310 315 320

AGT AAG GTA TGG AGA AAA GAT CTT TTA GAG TCA TCA TCA ATT CGC CGG 1008 Ser Lys Val Trp Arg Lys Asp Leu Leu Glu Ser Ser Serlie Arg Arg

325 330 335 CTC GCG GAG ATT ATA GTG AGT AAA TGT GGA GGA TTG CCA CTA GCG TTG 1056

Leu Ala Glu lie lie Val Ser Lys Cys Gly Gly Leu Pro Leu Ala Leu

340 345 350

ATC ACT TTA GGA GGA GCC ATG GCT 1080 lie Thr Leu Gly Gly Ala Met Ala

355 360

Claims

The scope of the claims

1. A DNA encoding a chimeric protein comprising at least a domain containing at least an elicitor binding site of an elicitor receptor and at least a domain containing a signal transduction motif of a protein that causes plant disease resistance.

2. The method according to claim 1, wherein the elicitor is an AVr gene product derived from glucan, polygalacturonic acid, N-acetylchitooligosaccharide, ericitin, Cladosporium fulvum, or a nip1 gene product derived from Rincosporium secalis. DN A.

3. The DNA according to claim 1, wherein the elicitor-binding site comprises the following recombinant protein (a) or (b).

(a) a protein consisting of the amino acid sequence represented by SEQ ID NO: 27

(b) a protein comprising an amino acid sequence represented by SEQ ID NO: 27 in which at least one amino acid has been deleted, substituted, added or inserted, and which has binding ability to glucan elicitor

4. The domain according to any one of claims 1 to 3, wherein the domain containing the signal transduction motif contains at least one selected from the group consisting of leucine-rich repeat, leucine zipper, nucleic acid binding site, and serine Z threonine kinase domain. The DNA according to any one of the above items.

5. The domain containing the signal transduction motif is the signal transduction domain of the expression product of the Pto gene, Prf gene, Cf-2 gene or Cf-9 gene derived from tomato, and the Xa21 gene derived from rice. Signaling domain of expression product, Signaling domain of expression product of RPS2 gene or RPM1 gene from Arabidopsis thaliana, Signaling domain of expression product of L6 gene from flax or signal of expression product of N gene from tobacco The DNA according to any one of claims 1 to 4, which is a transduction domain.

6. The DNA according to claim 5, wherein the RPS2 gene comprises the nucleotide sequence represented by SEQ ID NO: 29.

7. DNA encoding the following protein (c) or (d): (c) a chimeric protein comprising the amino acid sequence represented by SEQ ID NO: 5, 7, or 9

(d) a chimeric protein which comprises an amino acid sequence in which at least one amino acid is deleted, substituted, added or inserted in the amino acid sequence represented by SEQ ID NO: 5, 7 or 9, and which produces plant disease resistance

8. A DNA encoding a chimeric protein having plant disease resistance, comprising a nucleotide sequence represented by SEQ ID NO: 6, 8 or 10.

9. A recombinant vector containing the DNA of any one of claims 1 to 8.

10. A transformed plant transformed by the recombinant vector according to claim 9, or a progeny thereof.

1 1. A method for producing a disease-resistant plant, comprising introducing the recombinant vector according to claim 9 into a plant.

1 2. A plant disease-resistant plant or progeny thereof, into which the DNA of any one of claims 1 to 8 has been introduced and expressed.

1 3. A method for producing a plant having plant disease resistance or its progeny by integrating and expressing the DNA according to any one of claims 1 to 8 in a plant chromosome.