CN114736880A - Mutant D497N of glucose oxidase GoxM10 with improved acid stability as well as derivative mutant and application thereof - Google Patents
Mutant D497N of glucose oxidase GoxM10 with improved acid stability as well as derivative mutant and application thereof Download PDFInfo
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- CN114736880A CN114736880A CN202210648724.1A CN202210648724A CN114736880A CN 114736880 A CN114736880 A CN 114736880A CN 202210648724 A CN202210648724 A CN 202210648724A CN 114736880 A CN114736880 A CN 114736880A
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- 108010015776 Glucose oxidase Proteins 0.000 title claims abstract description 59
- 239000004366 Glucose oxidase Substances 0.000 title claims abstract description 55
- 229940116332 glucose oxidase Drugs 0.000 title claims abstract description 55
- 235000019420 glucose oxidase Nutrition 0.000 title claims abstract description 55
- 239000002253 acid Substances 0.000 title claims abstract description 41
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 22
- 102000004190 Enzymes Human genes 0.000 claims description 19
- 108090000790 Enzymes Proteins 0.000 claims description 19
- 229940088598 enzyme Drugs 0.000 claims description 18
- 230000035772 mutation Effects 0.000 claims description 18
- 239000013598 vector Substances 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 10
- 239000002773 nucleotide Substances 0.000 claims description 10
- 125000003729 nucleotide group Chemical group 0.000 claims description 10
- 239000003674 animal food additive Substances 0.000 claims description 2
- 230000001939 inductive effect Effects 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 9
- 235000013373 food additive Nutrition 0.000 claims 1
- 239000002778 food additive Substances 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000005516 engineering process Methods 0.000 abstract description 2
- 239000000463 material Substances 0.000 abstract description 2
- 230000001737 promoting effect Effects 0.000 abstract description 2
- 102000004169 proteins and genes Human genes 0.000 abstract description 2
- 238000011160 research Methods 0.000 abstract 2
- 150000001413 amino acids Chemical group 0.000 description 23
- 108020004414 DNA Proteins 0.000 description 21
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Abstract
Description
技术领域technical field
本发明属于饲料生物技术领域，具体涉及酸稳定性提高葡萄糖氧化酶GoxM10的突变体D497N及其衍生突变体和应用。The invention belongs to the technical field of feed biotechnology, and in particular relates to a mutant D497N of glucose oxidase GoxM10 with improved acid stability, a derivative mutant thereof, and applications thereof.
背景技术Background technique
葡萄糖氧化酶（Gox；EC1.1.3.4）是一种特性良好的需氧脱氢酶，在有氧条件下利用氧分子作为电子受体，催化β-d-葡萄糖生成d-葡萄糖酸内酯和过氧化氢，随后d-葡萄糖酸内酯水解成葡萄糖酸和水。近年来Gox作为一种新型抗生素替代添加剂受到饲料行业的普遍关注。Gox作为抗生素和生长促进剂的替代品，在不损害人类健康的前提下对保证动物健康和生产性能至关重要。大量应用研究表明，Gox作为一种绿色饲料添加剂，可用于预防牲畜胃肠道感染和腹泻，并有促进动物生长的作用。Glucose oxidase (Gox; EC1.1.3.4) is a well-characterized aerobic dehydrogenase that catalyzes the formation of d-gluconolactone from β-d-glucose using oxygen molecules as electron acceptors under aerobic conditions and hydrogen peroxide, followed by hydrolysis of d-gluconolactone to gluconic acid and water. In recent years, Gox as a new type of antibiotic substitute additive has received widespread attention in the feed industry. As an alternative to antibiotics and growth promoters, Gox is critical to animal health and performance without compromising human health. A large number of applied studies have shown that Gox, as a green feed additive, can be used to prevent gastrointestinal infections and diarrhea in livestock, and has the effect of promoting animal growth.
Gox能够在酸性环境中维持较高的活性，以便在动物胃肠道发挥最大作用是至关重要。所以为了满足Gox在饲料工业中的应用，Gox要具备良好的酸稳定性。现有技术已提高黑曲霉（Aspergillus niger）来源Gox的稳定性，获得了T 50增加7.5 ℃的突变体GoxM4，以及比GoxM4的T m提高了9 ℃的突变体GoxM10。The ability of Gox to maintain high activity in an acidic environment in order to exert its maximum effect in the gastrointestinal tract of animals is critical. Therefore, in order to meet the application of Gox in the feed industry, Gox must have good acid stability. The existing technology has improved the stability of Gox derived from Aspergillus niger , and obtained mutant GoxM4 with T 50 increased by 7.5 °C, and mutant GoxM10 with T m increased by 9 °C compared with GoxM4.
发明内容SUMMARY OF THE INVENTION
本发明的目的是提供酸稳定性提高的葡萄糖氧化酶GoxM10的突变体。It is an object of the present invention to provide mutants of glucose oxidase GoxM10 with improved acid stability.
本发明的再一目的是提供编码上述突变体的基因。Still another object of the present invention is to provide genes encoding the above mutants.
本发明的再一目的是提供包含上述突变体基因的重组载体。Still another object of the present invention is to provide a recombinant vector comprising the above mutant gene.
本发明的再一目的是提供包含上述突变体基因的重组菌株。Still another object of the present invention is to provide a recombinant strain comprising the above mutant gene.
本发明的再一目的是提供上述突变体的应用。Still another object of the present invention is to provide applications of the above mutants.
本发明的再一目的是提供制备稳定性提高的葡萄糖氧化酶的方法。Yet another object of the present invention is to provide a method for preparing a glucose oxidase with improved stability.
本发明的再一目的是提供提高葡萄糖氧化酶GoxM10的方法。Still another object of the present invention is to provide a method for increasing the glucose oxidase GoxM10.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶进行定点突变。According to a specific embodiment of the present invention, the glucose oxidase whose amino acid sequence is shown in SEQ ID NO: 1 is subjected to site-directed mutation.
根据本发明的具体实施方式，将氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶进行113、193、362、497、535、113/497、193/497、362/497、497/535氨基酸位点的突变，从而获得酸稳定性提高的葡萄糖氧化酶突变体。According to a specific embodiment of the present invention, the glucose oxidase whose amino acid sequence is shown in SEQ ID NO: 1 is subjected to 113, 193, 362, 497, 535, 113/497, 193/497, 362/497, 497/535 amino acids site to obtain a glucose oxidase mutant with improved acid stability.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶GoxM10进行H113S突变，得到氨基酸序列如SEQ ID NO:2所示的酸稳定提高的突变体。According to a specific embodiment of the present invention, H113S mutation is performed on the glucose oxidase GoxM10 whose amino acid sequence is shown in SEQ ID NO: 1 to obtain a mutant whose amino acid sequence is shown in SEQ ID NO: 2 with improved acid stability.
根据本发明的具体实施方式，还提供了编码上述酸稳定性提高的葡萄糖氧化酶突变体H113S的基因，其核苷酸序列如SEQ ID NO:3所示。According to a specific embodiment of the present invention, there is also provided a gene encoding the above-mentioned acid stability-improved glucose oxidase mutant H113S, the nucleotide sequence of which is shown in SEQ ID NO:3.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶GoxM10进行D193N突变，得到氨基酸序列如SEQ ID NO:4所示的酸稳定性提高的突变体。According to a specific embodiment of the present invention, the D193N mutation is performed on the glucose oxidase GoxM10 whose amino acid sequence is shown in SEQ ID NO: 1 to obtain a mutant whose amino acid sequence is shown in SEQ ID NO: 4 with improved acid stability.
根据本发明的具体实施方式，还提供了编码上述酸稳定性提高的葡萄糖氧化酶突变体D193N的基因，其核苷酸序列如SEQ ID NO:5所示。According to a specific embodiment of the present invention, there is also provided a gene encoding the above-mentioned acid stability-improved glucose oxidase mutant D193N, the nucleotide sequence of which is shown in SEQ ID NO:5.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶GoxM10进行K362Q突变，得到氨基酸序列如SEQ ID NO:6所示的酸稳定性提高的突变体。According to a specific embodiment of the present invention, K362Q mutation is performed on the glucose oxidase GoxM10 whose amino acid sequence is shown in SEQ ID NO: 1 to obtain a mutant whose amino acid sequence is shown in SEQ ID NO: 6 with improved acid stability.
根据本发明的具体实施方式，还提供了编码上述酸稳定性提高的葡萄糖氧化酶突变体K362Q的基因，其核苷酸序列如SEQ ID NO:7所示。According to a specific embodiment of the present invention, there is also provided a gene encoding the above-mentioned acid stability-improved glucose oxidase mutant K362Q, the nucleotide sequence of which is shown in SEQ ID NO:7.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶GoxM10进行D497N突变，得到氨基酸序列如SEQ ID NO:8所示的酸稳定性提高的突变体。According to a specific embodiment of the present invention, the D497N mutation is performed on the glucose oxidase GoxM10 whose amino acid sequence is shown in SEQ ID NO: 1 to obtain a mutant whose amino acid sequence is shown in SEQ ID NO: 8 with improved acid stability.
根据本发明的具体实施方式，还提供了编码上述酸稳定性提高的葡萄糖氧化酶突变体D497N的基因，其核苷酸序列如SEQ ID NO:9所示。According to a specific embodiment of the present invention, a gene encoding the above-mentioned acid stability-improved glucose oxidase mutant D497N is also provided, the nucleotide sequence of which is shown in SEQ ID NO: 9.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶GoxM10进行R535N突变，得到氨基酸序列如SEQ ID NO:10所示的酸稳定性提高的突变体。According to a specific embodiment of the present invention, R535N mutation is performed on the glucose oxidase GoxM10 whose amino acid sequence is shown in SEQ ID NO: 1 to obtain a mutant whose amino acid sequence is shown in SEQ ID NO: 10 with improved acid stability.
根据本发明的具体实施方式，还提供了编码上述酸稳定性提高的葡萄糖氧化酶突变体R535N的基因，其核苷酸序列如SEQ ID NO:11所示。According to a specific embodiment of the present invention, a gene encoding the above-mentioned glucose oxidase mutant R535N with improved acid stability is also provided, the nucleotide sequence of which is shown in SEQ ID NO: 11.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶GoxM10进行H113S/D497N突变，得到氨基酸序列如SEQ ID NO:12所示的酸稳定性提高的突变体。According to a specific embodiment of the present invention, H113S/D497N mutation is performed on the glucose oxidase GoxM10 whose amino acid sequence is shown in SEQ ID NO: 1 to obtain a mutant whose amino acid sequence is shown in SEQ ID NO: 12 with improved acid stability.
根据本发明的具体实施方式，还提供了编码上述酸稳定性提高的葡萄糖氧化酶突变体H113S/D497N的基因，其核苷酸序列如SEQID NO:13所示。According to a specific embodiment of the present invention, a gene encoding the above-mentioned acid stability-improved glucose oxidase mutant H113S/D497N is also provided, the nucleotide sequence of which is shown in SEQ ID NO: 13.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶GoxM10进行D193N/D497N突变，得到氨基酸序列如SEQ ID NO:14所示的酸稳定性提高的突变体。According to a specific embodiment of the present invention, D193N/D497N mutation is performed on the glucose oxidase GoxM10 whose amino acid sequence is shown in SEQ ID NO: 1 to obtain a mutant whose amino acid sequence is shown in SEQ ID NO: 14 with improved acid stability.
根据本发明的具体实施方式，还提供了编码上述酸稳定性提高的葡萄糖氧化酶突变体D193N/D497N的基因，其核苷酸序列如SEQ ID NO:15所示。According to a specific embodiment of the present invention, there is also provided a gene encoding the above-mentioned acid stability-improved glucose oxidase mutant D193N/D497N, the nucleotide sequence of which is shown in SEQ ID NO: 15.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶GoxM10进行K362Q/D497N突变，得到氨基酸序列如SEQ ID NO:16所示的酸稳定性提高的突变体。According to a specific embodiment of the present invention, K362Q/D497N mutation is performed on the glucose oxidase GoxM10 whose amino acid sequence is shown in SEQ ID NO: 1 to obtain a mutant whose amino acid sequence is shown in SEQ ID NO: 16 with improved acid stability.
根据本发明的具体实施方式，还提供了编码上述酸稳定性提高的葡萄糖氧化酶突变体K362Q/D497N的基因，其核苷酸序列如SEQ ID NO:17所示。According to a specific embodiment of the present invention, there is also provided a gene encoding the above-mentioned acid stability-improved glucose oxidase mutant K362Q/D497N, the nucleotide sequence of which is shown in SEQ ID NO: 17.
根据本发明的具体实施方式，对氨基酸序列如SEQ ID NO:1所示的葡萄糖氧化酶GoxM10进行D497N/R535N突变，得到氨基酸序列如SEQ ID NO:18所示的酸稳定性提高的突变体。According to a specific embodiment of the present invention, D497N/R535N mutation is performed on the glucose oxidase GoxM10 whose amino acid sequence is shown in SEQ ID NO: 1 to obtain a mutant whose amino acid sequence is shown in SEQ ID NO: 18 with improved acid stability.
根据本发明的具体实施方式，还提供了编码上述酸稳定性提高的葡萄糖氧化酶突变体D497N/R535N的基因，其核苷酸序列如SEQ ID NO:19所示。According to a specific embodiment of the present invention, a gene encoding the above-mentioned acid stability-improved glucose oxidase mutant D497N/R535N is also provided, the nucleotide sequence of which is shown in SEQ ID NO: 19.
根据本发明的具体实施方式，还提供了包含上述葡萄糖氧化酶突变体基因的重组载体，所述重组表达载体的出发载体为pPIC9，所述重组表达载体具体为pPIC9-goxm10-H113S，pPIC9-goxm10-D193N，pPIC9-goxm10-K362Q，pPIC9-goxm10-D497N，pPIC9-goxm10-R535N，pPIC9-goxm10-H113S/D497N，pPIC9-goxm10-D193N/D497N，pPIC9-goxm10-K362Q/D497N，pPIC9-goxm10-D497N/R535N。According to a specific embodiment of the present invention, there is also provided a recombinant vector comprising the above-mentioned glucose oxidase mutant gene, the starting vector of the recombinant expression vector is pPIC9, and the recombinant expression vector is specifically pPIC9- goxm10- H113S, pPIC9 -goxm10 -D193N, pPIC9 -goxm10- K362Q, pPIC9 -goxm10- D497N, pPIC9 -goxm10- R535N, pPIC9 -goxm10- H113S/D497N, pPIC9 -goxm10- D193N/D497N, pPIC9 -goxm10- K362Q/D49-D497N, pPIC9 -goxm10- D497N /R535N.
根据本发明的具体实施方式，还提供了包含上述葡萄糖氧化酶突变体基因的重组菌株，所述重组菌的出发菌株具体为GS115(pPIC9-goxm10)。According to a specific embodiment of the present invention, a recombinant strain comprising the above-mentioned glucose oxidase mutant gene is also provided, and the starting strain of the recombinant strain is specifically GS115 (pPIC9 -goxm10 ).
根据本发明的制备具有稳定性提高的葡萄糖氧化酶的方法，包括以下步骤：The method for preparing glucose oxidase with improved stability according to the present invention comprises the following steps:
1）制备包含上述突变体基因的重组载体；1) Prepare a recombinant vector comprising the above mutant gene;
2）所述重组载体转化毕赤酵母GS115表达宿主；2) The recombinant vector is transformed into the Pichia pastoris GS115 expression host;
3）发酵培养所述宿主，并分离葡萄糖氧化酶。3) The host is fermented and cultured, and glucose oxidase is isolated.
本发明提供了上述酸稳定性改进的葡萄糖氧化酶突变体的应用，具体可以应用于食品和饲料领域中。The present invention provides the application of the above-mentioned glucose oxidase mutant with improved acid stability, which can be specifically applied in the field of food and feed.
本发明的突变体与GoxM10相比，酸稳定性均明显提高。突变体D193N、D497N和D193N/D497N的最适pH降低1个单位，双点突变体H113S/D497N、K362Q/D497N和D497N/R535N的最适反应pH为5.5，与GoxM10相比下降了0.5个单位，其他突变体的最适pH和GoxM10一致均为6.0。pH 2.5条件下处理30 min时，突变体H113S、D193N、K362Q、D497N、R535N、H113S/D497N、D193N/D497N、K362Q/D497N、D497N/R535N的相对剩余酶活力分别为22%、26%、26%、34%、24%、27%、35%、25%、28%，高于GoxM10（21%）；在pH 2.75和37 ℃条件下处理60 min，突变体H113S、D193N、K362Q、D497N、R535N、H113S/D497N、D193N/D497N、K362Q/D497N、D497N/R535N的相对剩余酶活力分别为26%、28%、26%、32%、29%、47%、49%、32%、43%左右，高于GoxM10的相对剩余酶活力24%。Compared with GoxM10, the mutants of the present invention have significantly improved acid stability. The pH optimum of mutants D193N, D497N and D193N/D497N decreased by 1 unit, and the optimum pH of double point mutants H113S/D497N, K362Q/D497N and D497N/R535N was 5.5, a decrease of 0.5 units compared with GoxM10 , the optimum pH of other mutants is consistent with that of GoxM10 at 6.0. When treated at pH 2.5 for 30 min, the relative residual enzyme activities of mutants H113S, D193N, K362Q, D497N, R535N, H113S/D497N, D193N/D497N, K362Q/D497N, D497N/R535N were 22%, 26%, and 26%, respectively. %, 34%, 24%, 27%, 35%, 25%, 28%, higher than GoxM10 (21%); treated at pH 2.75 and 37 ℃ for 60 min, mutants H113S, D193N, K362Q, D497N, The relative residual enzyme activities of R535N, H113S/D497N, D193N/D497N, K362Q/D497N, D497N/R535N were 26%, 28%, 26%, 32%, 29%, 47%, 49%, 32%, 43%, respectively about 24% higher than the relative residual enzyme activity of GoxM10.
本发明提供的葡萄糖氧化酶突变体能很好的满足食品和饲料等领域中的应用，有着非常广阔的应用前景。The glucose oxidase mutant provided by the invention can well meet the application in the fields of food and feed, and has a very broad application prospect.
附图说明Description of drawings
图1显示GoxM10和突变体在pH 2.5条件下处理30 min的酸稳定性对比；Figure 1 shows a comparison of the acid stability of GoxM10 and mutants treated at pH 2.5 for 30 min;
图2显示GoxM10和突变体在pH 2.75条件下处理60 min的酸稳定性对比；Figure 2 shows a comparison of the acid stability of GoxM10 and mutants treated at pH 2.75 for 60 min;
图3显示GoxM10和突变体的最适pH对比。Figure 3 shows a comparison of pH optimum for GoxM10 and mutants.
具体实施方式Detailed ways
试验材料和试剂Test Materials and Reagents
1、菌株及载体：表达宿主为Pichia pastoris GS115，表达质粒载体为pPIC9。1. Strain and vector: The expression host is Pichia pastoris GS115, and the expression plasmid vector is pPIC9.
2、酶类及其它生化试剂：内切酶/高保真DNA聚合酶和重组酶。2. Enzymes and other biochemical reagents: endonuclease/high-fidelity DNA polymerase and recombinase.
3、大肠杆菌LB培养基（1%蛋白胨、0.5%酵母提取物、1% NaCl，pH自然）。毕赤酵母YPD培养基（1%酵母提取物，2%蛋白胨，2%葡萄糖，pH自然）；BMGY(1%酵母提取物，2%蛋白胨，10%无氨基酵母氮源，0.1%生物素，pH自然)；BMMY（1%酵母提取物，2%蛋白胨，10%无氨基酵母氮源，0.1%生物素，1%甲醇，pH自然）。3. Escherichia coli LB medium (1% peptone, 0.5% yeast extract, 1% NaCl, natural pH). Pichia YPD medium (1% yeast extract, 2% peptone, 2% glucose, pH natural); BMGY (1% yeast extract, 2% peptone, 10% amino-free yeast nitrogen source, 0.1% biotin, pH natural); BMMY (1% yeast extract, 2% peptone, 10% amino-free yeast nitrogen source, 0.1% biotin, 1% methanol, pH natural).
说明：以下实施例中未作具体说明的分子生物学实验方法，均参照《分子克隆实验指南》（第三版）J.萨姆布鲁克一书中所列的具体方法进行，或者按照试剂盒和产品说明书进行。Note: The molecular biology experimental methods that are not specifically described in the following examples are all carried out with reference to the specific methods listed in the book "Molecular Cloning Experiment Guide" (Third Edition) by J. Sambrook, or according to the kit and product manual.
实施例1、重组菌株GS115(pPIC9-goxm10)的制备Example 1. Preparation of recombinant strain GS115 (pPIC9 -goxm10 )
1. 扩增高热稳定性葡萄糖氧化酶GoxM10的核酸序列goxm10 1. Amplify the nucleic acid sequence goxm10 of the hyperthermostable glucose oxidase GoxM10
采用PCR的方法扩增goxm10基因片段，采用双酶切方法获得载体pPIC9核酸片段，通过重组酶将两者连接，获得重组质粒pPIC9-goxm10，并转化毕赤酵母GS115，获得重组毕赤酵母菌株GS115(pPIC9-goxm10)。PCR所用引物如下：The goxm10 gene fragment was amplified by PCR, the nucleic acid fragment of vector pPIC9 was obtained by double enzyme digestion, and the two were connected by recombinase to obtain the recombinant plasmid pPIC9- goxm10 , and transformed into Pichia pastoris GS115 to obtain recombinant Pichia pastoris strain GS115 (pPIC9 -goxm10 ). The primers used for PCR are as follows:
GoxM10-F（SEQ ID NO: 20），GoxM10-R（SEQ ID NO: 21）。GoxM10-F (SEQ ID NO: 20), GoxM10-R (SEQ ID NO: 21).
其中，GoxM10-F及GoxM10-R用于扩增GoxM10的基因编码序列；载体pPIC9通过将保存的菌种接到液体LB培养基过夜培养后，质粒提取得到。Among them, GoxM10-F and GoxM10-R are used to amplify the gene coding sequence of GoxM10; the vector pPIC9 is obtained by culturing the preserved strain in liquid LB medium overnight, and then extracting the plasmid.
扩增结束后，将PCR产物以及提取质粒进行核酸电泳检测，goxm10与载体pPIC9条带大小分别为1746bp、8088bp，将载体用EcoRI和NotI进行酶切后，将PCR产物与酶切产物分别回收纯化。After the amplification, the PCR product and the extracted plasmid were detected by nucleic acid electrophoresis. The band sizes of goxm10 and the carrier pPIC9 were respectively 1746bp and 8088bp. After the vector was digested with Eco RI and Not I, the PCR product and the digested product were respectively Recovery and purification.
2. 构建重组菌株GS115(pPIC9-goxm10)2. Construction of recombinant strain GS115 (pPIC9 -goxm10 )
将回收的goxm10与pPIC9基因片段通过重组酶进行重组连接，然后将重组产物转化大肠杆菌JM109感受态，并涂布于LB（含100 μg/mL Amp）进行筛选。待测序正确后，利用BglⅡ限制性内切酶将重组质粒pPIC9-goxm10进行酶切，回收酶切产物后电击转化毕赤酵母GS115感受态细胞，得到重组表达菌株GS115(pPIC9-goxm10)。The recovered goxm10 and pPIC9 gene fragments were recombined and ligated by recombinase, and then the recombinant product was transformed into E. coli JM109 competent and spread on LB (containing 100 μg/mL Amp) for screening. After the sequencing was correct, the recombinant plasmid pPIC9 -goxm10 was digested with Bgl II restriction enzyme, and the digested product was recovered and electroporated into Pichia pastoris GS115 competent cells to obtain recombinant expression strain GS115 (pPIC9 -goxm10 ).
实施例2、重组突变菌株GS115的制备Example 2. Preparation of recombinant mutant strain GS115
1. 稳定性提高的重组质粒的构建1. Construction of recombinant plasmids with improved stability
根据突变体设计突变引物，以质粒pPIC9-goxm10为模板，通过点突变试剂盒引入突变氨基酸，含突变氨基酸的PCR产物用DpnⅠ消化处理以去除模板，消化后的PCR产物转化进入大肠杆菌JM109感受态细胞，并对其进行测序验证，获得含葡萄糖氧化酶突变质粒的大肠杆菌pPIC9-goxm10-H113S，pPIC9-goxm10-D193N，pPIC9-goxm10-K362Q，pPIC9-goxm10-D497N，pPIC9-goxm10-R535N， pPIC9-goxm10-H113S/D497N，pPIC9-goxm10-D193N/D497N，pPIC9-goxm10-K362Q/D497N，pPIC9-goxm10-D497N/R535N。所用引物如下所示：Mutation primers were designed according to the mutants. Using plasmid pPIC9- goxm10 as the template, mutant amino acids were introduced by a point mutation kit. The PCR products containing the mutant amino acids were digested with Dpn I to remove the template. The digested PCR products were transformed into E. coli JM109 The cells were sequenced and verified to obtain E. coli pPIC9 -goxm10- H113S, pPIC9 -goxm10- D193N, pPIC9 -goxm10- K362Q, pPIC9 -goxm10- D497N, pPIC9 -goxm10 -R535N containing glucose oxidase mutant plasmids, pPIC9 -goxm10- H113S/D497N, pPIC9 -goxm10- D193N/D497N, pPIC9 -goxm10- K362Q/D497N, pPIC9 -goxm10- D497N/R535N. The primers used are as follows:
Gox-H113S-F（SEQ ID NO：22），Gox-H113S-R（SEQ ID NO：23）；Gox-D193N-F（SEQID NO：24），Gox-D193N-R（SEQ ID NO：25）；Gox-K362Q-F（SEQ ID NO：26），Gox-K362Q-R（SEQID NO：27）；Gox-D497N-F（SEQ ID NO：28），Gox-D497N-R（SEQ ID NO：29）；Gox-H113S-F (SEQ ID NO:22), Gox-H113S-R (SEQ ID NO:23); Gox-D193N-F (SEQ ID NO:24), Gox-D193N-R (SEQ ID NO:25) ; Gox-K362Q-F (SEQ ID NO: 26), Gox-K362Q-R (SEQ ID NO: 27); Gox-D497N-F (SEQ ID NO: 28), Gox-D497N-R (SEQ ID NO: 29 );
Gox-R535N-F（SEQ ID NO：30），Gox-R535N-R（SEQ ID NO：31）。Gox-R535N-F (SEQ ID NO:30), Gox-R535N-R (SEQ ID NO:31).
2. 酸稳定性和热稳定重组菌株GS115构建2. Construction of acid-stable and thermostable recombinant strain GS115
利用BglⅡ将测序正确的突变质粒进行酶切，回收酶切产物后电击转化毕赤酵母GS115感受态细胞，得到重组表达菌株GS115(pPIC9-goxm10-H113S)，GS115(pPIC9-goxm10-D193N)，GS115(pPIC9-goxm10-K362Q)，GS115(pPIC9-goxm10-D497N)，GS115(pPIC9-goxm10-R535N)， GS115(pPIC9-goxm10-H113S/D497N)，GS115(pPIC9-goxm10-D193N/D497N)，GS115(pPIC9-goxm10-K362Q/D497N)，GS115(pPIC9-goxm10-D497N/R535N)。The correctly sequenced mutant plasmid was digested with Bgl II, and the digested product was recovered and electroporated into Pichia pastoris GS115 competent cells to obtain recombinant expression strains GS115 (pPIC9 -goxm10- H113S), GS115 (pPIC9 -goxm10- D193N), GS115(pPIC9 -goxm10- K362Q), GS115(pPIC9 -goxm10- D497N), GS115(pPIC9 -goxm10- R535N), GS115(pPIC9 -goxm10- H113S/D497N), GS115(pPIC9 -goxm10- D193N/D4957N), GS1 (pPIC9 -goxm10- K362Q/D497N), GS115 (pPIC9 -goxm10- D497N/R535N).
实施例3、葡萄糖氧化酶GoxM10及高酸稳定性和热稳定性突变体酶蛋白的获得Example 3. Acquisition of Glucose Oxidase GoxM10 and High Acid Stability and Thermostable Mutant Enzyme Proteins
1. GoxM10及突变体的诱导表达1. Inducible expression of GoxM10 and mutants
以1%的接种量将上述得到的重组酵母表达菌株接种至50 mL YPD培养基中进行种子液培养，200 rpm，30 ℃培养48 h后，以1%接种量转接至400 mL BMGY培养基中，200 rpm，30 ℃培养48 h，离心收集菌体转接到200 mL BMMY培养基中，每24 h补加1%的甲醇进行诱导表达，共培养72 h。The recombinant yeast expression strain obtained above was inoculated into 50 mL of YPD medium with an inoculum of 1% for seed liquid culture, incubated at 200 rpm at 30 °C for 48 h, and then transferred to 400 mL of BMGY medium with an inoculum of 1%. The cells were cultured at 200 rpm and 30 °C for 48 h. The cells were collected by centrifugation and transferred to 200 mL of BMMY medium. 1% methanol was added every 24 h to induce expression for a total of 72 h.
2. GoxM10及突变体的纯化2. Purification of GoxM10 and mutants
将诱导表达后的菌液12000 rpm, 10 min离心，收集上清后用10 kDa的膜包进行浓缩，再用10 mM磷酸氢二钠-柠檬酸缓冲液（pH 6.5）进行透析，然后将透析后的粗酶液用阴离子柱纯化，纯化用A液为10 mM磷酸氢二钠-柠檬酸缓冲液（pH 6.5），B液为A液加1 MNaCl，纯化蛋白，收集洗脱液。Centrifuge the induced expression at 12,000 rpm for 10 min, collect the supernatant and concentrate it with a 10 kDa membrane bag, then dialyze it with 10 mM disodium hydrogen phosphate-citric acid buffer (pH 6.5), and then dialyze the The crude enzyme solution was purified by an anion column. The purification solution A was 10 mM disodium hydrogen phosphate-citric acid buffer (pH 6.5), and the B solution was solution A plus 1 M NaCl to purify the protein and collect the eluate.
3. GoxM10和突变体的酶学性质测定3. Determination of the enzymatic properties of GoxM10 and mutants
（1）GoxM10和突变体的酸稳定性测定(1) Acid stability assay of GoxM10 and mutants
1采用两种测定方法评估GoxM10和突变体的pH稳定性。方法一：把相同浓度的GoxM10和突变体酶液置于准确配置的pH 2.5、2.75、3.0、4.0、5.0、6.0、7.0和8.0的0.1 M的磷酸氢二钠-柠檬酸缓冲液，然后于37 ℃保温处理1 h。保温结束后在标准条件下测定其剩余酶活力，以未处理样品的酶活力记为100%。方法二：把相同浓度的GoxM10和突变体酶液置于pH 2.5的0.1 M的磷酸氢二钠-柠檬酸缓冲液，然后于37 ℃保温处理10 min、20 min、30min、40 min、50 min和60 min。保温结束后在标准条件下测定GoxM10及其突变体酶液的相对剩余酶活。如图1所示，pH 2.5条件下处理30 min时，突变体H113S、D193N、K362Q、D497N、R535N、H113S/D497N、D193N/D497N、K362Q/D497N、D497N/R535N的相对剩余酶活力分别为22%、26%、26%、34%、24%、27%、35%、25%、28%，与GoxM10（21%）相比分别提高了约5%、24%、24%、62%、14%、29%、67%、19%、33%。如图2所示，在pH 2.75和37 ℃条件下处理60 min，突变体H113S、D193N、K362Q、D497N、R535N、H113S/D497N、D193N/D497N、K362Q/D497N、D497N/R535N的相对剩余酶活力分别为26%、28%、26%、32%、29%、47%、49%、32%、43%左右，与GoxM10（24%）相比分别提高了约8%、17%、8%、33%、21%、96%、104%、33%、79%。1 Two assays were used to assess the pH stability of GoxM10 and mutants. Method 1: Put the same concentration of GoxM10 and mutant enzyme solution in 0.1 M disodium hydrogen phosphate-citric acid buffer at pH 2.5, 2.75, 3.0, 4.0, 5.0, 6.0, 7.0 and 8.0, and then Incubate at 37 °C for 1 h. After the incubation, the remaining enzyme activity was measured under standard conditions, and the enzyme activity of the untreated sample was recorded as 100%. Method 2: Put the same concentration of GoxM10 and mutant enzyme solution in 0.1 M disodium hydrogen phosphate-citric acid buffer at pH 2.5, and then incubate at 37 °C for 10 min, 20 min, 30 min, 40 min, 50 min and 60 min. After the incubation, the relative residual enzyme activities of GoxM10 and its mutant enzyme solutions were determined under standard conditions. As shown in Figure 1, the relative residual enzyme activities of mutants H113S, D193N, K362Q, D497N, R535N, H113S/D497N, D193N/D497N, K362Q/D497N, D497N/R535N were 22 when treated at pH 2.5 for 30 min, respectively. %, 26%, 26%, 34%, 24%, 27%, 35%, 25%, 28%, which were increased by about 5%, 24%, 24%, 62%, 14%, 29%, 67%, 19%, 33%. As shown in Figure 2, the relative residual enzyme activities of mutants H113S, D193N, K362Q, D497N, R535N, H113S/D497N, D193N/D497N, K362Q/D497N, D497N/R535N after treatment at pH 2.75 and 37 °C for 60 min They are about 26%, 28%, 26%, 32%, 29%, 47%, 49%, 32%, and 43%, respectively, which are about 8%, 17%, and 8% higher than GoxM10 (24%). , 33%, 21%, 96%, 104%, 33%, 79%.
（4）GoxM10和突变体的最适pH测(4) Optimal pH measurement of GoxM10 and mutants
首先，分别配置不同pH梯度（3.0、4.0、5.0、6.0、7.0、8.0）的Gox酶活力测定的底物混合液，然后，在30 ℃下测定GoxM10及其突变体的酶活力，把最适反应pH条件下的酶活力设定为100%，然后计算其余pH条件下的相对酶活力，即可得到GoxM10及其突变体的最适反应pH。如图3所示，突变体D193N、D497N和D193N/D497N的最适pH为5，与GoxM10相比下降一个单位，突变体H113S/D497N、K362Q/D497N和D497N/R535N的最适反应pH为5.5，与GoxM10相比下降了0.5个单位，突变体H113S、K362Q、R535N的最适反应pH与GoxM10相比均为6.0。First, the substrate mixtures for the determination of Gox enzyme activity with different pH gradients (3.0, 4.0, 5.0, 6.0, 7.0, 8.0) were prepared respectively, and then the enzyme activities of GoxM10 and its mutants were measured at 30 °C, and the optimal The enzyme activity under the reaction pH condition was set to 100%, and then the relative enzyme activity under other pH conditions was calculated to obtain the optimal reaction pH of GoxM10 and its mutants. As shown in Figure 3, the optimum pH of mutants D193N, D497N and D193N/D497N was 5, which was one unit lower than that of GoxM10, and the optimum pH of mutants H113S/D497N, K362Q/D497N and D497N/R535N was 5.5 , compared with GoxM10, decreased by 0.5 unit, and the optimal pH of mutant H113S, K362Q, R535N was 6.0 compared with that of GoxM10.
以上实施例仅用于解释本申请的技术方案，不限定本申请的保护范围。The above embodiments are only used to explain the technical solutions of the present application, and do not limit the protection scope of the present application.
序列表 sequence listing
<110> 中国农业科学院北京畜牧兽医研究所<110> Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences
<120> 酸稳定性提高葡萄糖氧化酶GoxM10的突变体D497N及其衍生突变体和应用<120> Mutant D497N of Glucose Oxidase GoxM10 with Improved Acid Stability and Its Derived Mutants and Applications
<160> 31<160> 31
<170> SIPOSequenceListing 1.0<170> SIPOSequenceListing 1.0
<210> 1<210> 1
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 1<400> 1
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
His Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlyHis Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Gln Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnGln Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 2<210> 2
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 2<400> 2
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
Ser Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlySer Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Ser Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnSer Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 3<210> 3
<211> 1746<211> 1746
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 3<400> 3
ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60
atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120
gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180
gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240
gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300
tctactttgg ttaacggagg tacttggacc agaccatcta aggctcaagt tgactcttgg 360tctactttgg ttaacggagg tacttggacc agaccatcta aggctcaagt tgactcttgg 360
gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420
gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480
tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540
tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600
gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660
caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720
caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780
agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840
cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900
atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960
ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020
gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080
gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140
agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200
attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260
ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320
ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380
ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440
tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500
tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560
atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620
ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680
gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740
caataa 1746caataa 1746
<210> 4<210> 4
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 4<400> 4
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
His Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlyHis Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asn Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsn Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Ser Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnSer Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 5<210> 5
<211> 1746<211> 1746
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 5<400> 5
ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60
atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120
gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180
gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240
gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300
tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360
gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420
gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480
tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540
tctcctatcg tcaaggcctt gatgtccgct gttgaaaaca gaggtgtccc aactaagaag 600tctcctatcg tcaaggcctt gatgtccgct gttgaaaaca gaggtgtccc aactaagaag 600
gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660
caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720
caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780
agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840
cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900
atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960
ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020
gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080
gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140
agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200
attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260
ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320
ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380
ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440
tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500
tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560
atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620
ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680
gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740
caataa 1746caataa 1746
<210> 6<210> 6
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 6<400> 6
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
His Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlyHis Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Ser Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnSer Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Gln Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Gln Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 7<210> 7
<211> 1746<211> 1746
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 7<400> 7
ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60
atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120
gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180
gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240
gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300
tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360
gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420
gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480
tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540
tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600
gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660
caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720
caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780
agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840
cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900
atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960
ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020
gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080
gagcaagctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140gagcaagctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140
agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200
attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260
ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320
ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380
ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440
tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500
tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560
atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620
ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680
gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740
caataa 1746caataa 1746
<210> 8<210> 8
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 8<400> 8
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
His Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlyHis Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Ser Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnSer Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 9<210> 9
<211> 1746<211> 1746
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 9<400> 9
ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60
atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120
gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180
gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240
gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300
tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360
gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420
gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480
tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540
tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600
gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660
caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720
caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780
agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840
cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900
atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960
ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020
gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080
gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140
agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200
attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260
ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320
ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380
ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440
tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctaa cttgagagct 1500tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctaa cttgagagct 1500
tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560
atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620
ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680
gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740
caataa 1746caataa 1746
<210> 10<210> 10
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 10<400> 10
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
His Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlyHis Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Ser Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnSer Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsp Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Asn Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Asn Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 11<210> 11
<211> 1746<211> 1746
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 11<400> 11
ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60
atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120
gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180
gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240
gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300
tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360
gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420
gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480
tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540
tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600
gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660
caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720
caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780
agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840
cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900
atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960
ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020
gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080
gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140
agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200
attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260
ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320
ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380
ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440
tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500
tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560
atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctaacgttta cggtgtccag 1620atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctaacgttta cggtgtccag 1620
ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680
gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740
caataa 1746caataa 1746
<210> 12<210> 12
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 12<400> 12
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
Ser Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlySer Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Ser Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnSer Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 13<210> 13
<211> 1746<211> 1746
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 13<400> 13
ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60
atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120
gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180
gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240
gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300
tctactttgg ttaacggagg tacttggacc agaccatcta aggctcaagt tgactcttgg 360tctactttgg ttaacggagg tacttggacc agaccatcta aggctcaagt tgactcttgg 360
gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420
gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480
tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540
tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600
gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660
caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720
caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780
agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840
cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900
atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960
ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020
gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080
gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140
agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200
attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260
ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320
ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380
ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440
tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctaa cttgagagct 1500tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctaa cttgagagct 1500
tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560
atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620
ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680
gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740
caataa 1746caataa 1746
<210> 14<210> 14
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 14<400> 14
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
His Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlyHis Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asn Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsn Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Ser Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnSer Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 15<210> 15
<211> 1746<211> 1746
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 15<400> 15
ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60
atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120
gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180
gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240
gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300
tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360
gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420
gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480
tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540
tctcctatcg tcaaggcctt gatgtccgct gttgaaaaca gaggtgtccc aactaagaag 600tctcctatcg tcaaggcctt gatgtccgct gttgaaaaca gaggtgtccc aactaagaag 600
gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660
caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720
caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780
agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840
cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900
atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960
ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020
gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080
gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140
agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200
attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260
ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320
ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380
ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440
tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctaa cttgagagct 1500tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctaa cttgagagct 1500
tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560
atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620
ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680
gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740
caataa 1746caataa 1746
<210> 16<210> 16
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 16<400> 16
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
His Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlyHis Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Ser Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnSer Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Gln Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Gln Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Arg Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 17<210> 17
<211> 1746<211> 1746
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 17<400> 17
ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60
atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120
gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180
gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240
gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300
tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360
gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420
gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480
tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540
tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600
gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660
caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720
caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780
agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840
cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900
atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960
ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020
gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080
gagcaagctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140gagcaagctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140
agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200
attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260
ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320
ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380
ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440
tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctaa cttgagagct 1500tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctaa cttgagagct 1500
tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560
atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctagagttta cggtgtccag 1620
ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680
gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740
caataa 1746caataa 1746
<210> 18<210> 18
<211> 581<211> 581
<212> PRT<212> PRT
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 18<400> 18
Gly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly ArgGly Ile Glu Ala Ser Leu Leu Thr Asp Pro Lys Glu Val Ala Gly Arg
1 5 10 151 5 10 15
Thr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val ValThr Val Asp Tyr Ile Ile Ala Gly Gly Gly Leu Thr Gly Leu Val Val
20 25 30 20 25 30
Ala Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile GluAla Ala Arg Leu Thr Glu Asn Pro Asp Ile Thr Val Leu Val Ile Glu
35 40 45 35 40 45
Ser Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu AsnSer Gly Ser Tyr Glu Ser Asp Arg Gly Pro Ile Ile Glu Asp Leu Asn
50 55 60 50 55 60
Ala Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu ThrAla Tyr Gly Lys Ile Phe Gly Ser Ser Val Asp His Ala Tyr Glu Thr
65 70 75 8065 70 75 80
Val Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly AsnVal Cys Leu Ala Thr Asn Asn Arg Thr Ala Leu Ile Arg Ala Gly Asn
85 90 95 85 90 95
Gly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg ProGly Leu Gly Gly Ser Thr Leu Val Asn Gly Gly Thr Trp Thr Arg Pro
100 105 110 100 105 110
His Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu GlyHis Lys Ala Gln Val Asp Ser Trp Glu Thr Val Phe Gly Asn Glu Gly
115 120 125 115 120 125
Trp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg AlaTrp Asn Trp Asp Ser Val Ala Ala Tyr Ser Leu Gln Ala Glu Arg Ala
130 135 140 130 135 140
Arg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn AlaArg Ala Pro Asn Ala Lys Gln Ile Ala Ala Gly His Tyr Phe Asn Ala
145 150 155 160145 150 155 160
Ser Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp ThrSer Cys His Gly Ile Asn Gly Thr Val His Ala Gly Pro Arg Asp Thr
165 170 175 165 170 175
Gly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val GluGly Asp Asp Tyr Ser Pro Ile Val Lys Ala Leu Met Ser Ala Val Glu
180 185 190 180 185 190
Asp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro HisAsp Arg Gly Val Pro Thr Lys Lys Asp Leu Gly Cys Gly Asp Pro His
195 200 205 195 200 205
Gly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg SerGly Val Ser Met Phe Pro Asn Thr Leu His Glu Asp Gln Val Arg Ser
210 215 220 210 215 220
Asp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn LeuAsp Ala Ala Arg Glu Trp Leu Leu Pro Asn Tyr Gln Arg Pro Asn Leu
225 230 235 240225 230 235 240
Ser Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln AsnSer Val Leu Thr Gly Gln Tyr Val Gly Lys Val Leu Leu Ser Gln Asn
245 250 255 245 250 255
Ala Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys GlyAla Thr Thr Pro Arg Ala Val Gly Val Glu Phe Gly Thr His Lys Gly
260 265 270 260 265 270
Asn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala GlyAsn Phe His Asn Val Thr Ala Lys His Glu Val Leu Leu Ala Ala Gly
275 280 285 275 280 285
Ser Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met LysSer Ala Val Ser Pro Thr Ile Leu Glu Tyr Ser Gly Ile Gly Met Lys
290 295 300 290 295 300
Ser Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro ValSer Ile Leu Glu Pro Leu Gly Ile Lys Thr Val Val Asp Leu Pro Val
305 310 315 320305 310 315 320
Gly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg IleGly Leu Asn Leu Gln Asp Gln Thr Thr Ser Thr Val Arg Ser Arg Ile
325 330 335 325 330 335
Thr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr PheThr Ser Ala Gly Ala Gly Gln Gly Gln Ala Ala Trp Phe Ala Thr Phe
340 345 350 340 345 350
Asn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu AsnAsn Glu Thr Phe Gly Asp Tyr Thr Glu Lys Ala His Glu Leu Leu Asn
355 360 365 355 360 365
Thr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly PheThr Lys Leu Glu Gln Trp Ala Glu Glu Ala Val Ala Arg Gly Gly Phe
370 375 380 370 375 380
His Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp TrpHis Asn Thr Thr Ala Leu Leu Ile Gln Tyr Glu Asn Tyr Arg Asp Trp
385 390 395 400385 390 395 400
Ile Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr AlaIle Val Lys Asp Asn Val Ala Tyr Ser Glu Leu Phe Leu Asp Thr Ala
405 410 415 405 410 415
Gly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg GlyGly Glu Ala Ser Phe Asp Val Trp Asp Leu Leu Pro Phe Thr Arg Gly
420 425 430 420 425 430
Tyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala TyrTyr Val His Ile Leu Asp Lys Asp Pro Tyr Leu Arg His Phe Ala Tyr
435 440 445 435 440 445
Asp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala AlaAsp Pro Gln Tyr Phe Leu Asn Glu Leu Asp Leu Leu Gly Gln Ala Ala
450 455 460 450 455 460
Ala Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln ThrAla Thr Gln Leu Ala Arg Asn Ile Ser Asn Ser Gly Ala Met Gln Thr
465 470 475 480465 470 475 480
Tyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp AlaTyr Phe Ala Gly Glu Thr Ile Pro Gly Asp Asn Leu Ala Tyr Asp Ala
485 490 495 485 490 495
Asn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro AsnAsn Leu Arg Ala Trp Val Glu Tyr Ile Pro Tyr His Phe Arg Pro Asn
500 505 510 500 505 510
Tyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly GlyTyr His Gly Val Gly Thr Cys Ser Met Met Pro Lys Glu Met Gly Gly
515 520 525 515 520 525
Val Val Asp Asn Ala Ala Asn Val Tyr Gly Val Gln Gly Leu Arg ValVal Val Asp Asn Ala Ala Asn Val Tyr Gly Val Gln Gly Leu Arg Val
530 535 540 530 535 540
Ile Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met ThrIle Asp Gly Ser Ile Pro Pro Thr Gln Met Ser Ser His Val Met Thr
545 550 555 560545 550 555 560
Val Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala AspVal Phe Tyr Ala Met Ala Leu Lys Ile Ala Asp Ala Val Leu Ala Asp
565 570 575 565 570 575
Tyr Ala Ser Met GlnTyr Ala Ser Met Gln
580 580
<210> 19<210> 19
<211> 1746<211> 1746
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 19<400> 19
ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60ggtattgagg cttccttgtt gactgaccca aaggaggtcg ccggtagaac tgttgactac 60
atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120atcattgctg gtggtggatt gactggtttg gttgtcgctg ccagattgac tgagaaccca 120
gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180gacatcaccg ttttggtcat tgagtccggt tcttacgaat ctgatagagg tcctatcatt 180
gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240gaagacttga acgcttacgg taaaatcttc ggatcttccg ttgaccacgc ttacgagact 240
gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300gtctgccttg ccactaacaa tagaaccgct ttgattagag ctggtaacgg tttgggtggt 300
tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360tctactttgg ttaacggagg tacttggacc agaccacaca aggctcaagt tgactcttgg 360
gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420gagaccgtct tcggtaacga aggttggaat tgggattctg tcgcagctta ctccttgcag 420
gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480gccgagagag cccgtgctcc aaacgctaag caaatcgccg caggtcacta cttcaacgcc 480
tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540tcctgtcacg gtattaacgg aactgttcac gctggtccaa gagacaccgg tgacgattac 540
tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600tctcctatcg tcaaggcctt gatgtccgct gttgaagaca gaggtgtccc aactaagaag 600
gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660gacttgggtt gcggagaccc acatggtgtt tctatgttcc ctaacacctt gcacgaggac 660
caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720caagtcagat ccgatgctgc ccgtgaatgg ttgcttccaa actaccaaag acctaacttg 720
caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780caggttttga ccggtcaata cgttggtaag gtccttttgt ctcaaaacgc cactacccca 780
agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840agagctgttg gtgtcgagtt cggaactcac aagggtaact ttcacaatgt taccgctaaa 840
cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900cacgaagtcc ttttggcagc tggttccgct gtttctccaa ctatcttgga gtactctggt 900
atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960atcggaatga agtccatttt ggaaccactt ggtattaaga ccgtcgttga cttgcctgtt 960
ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020ggtctgaact tgcaagacca gactacctct actgtcagat cccgtattac ctccgccggt 1020
gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080gctggacagg gtcaggctgc ctggtttgct actttcaacg agaccttcgg tgactacact 1080
gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140gagaaggctc acgaattgct taacaccaaa ttggaacaat gggctgagga agccgttgct 1140
agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200agaggtggtt tccacaacac taccgctctt ttgatccaat acgagaacta cagagactgg 1200
attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260attgttaagg ataacgtcgc ttactctgaa ttgttcttgg acactgccgg tgaggcttcc 1260
ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320ttcgacgtct gggacttgct gccattcact agaggatacg ttcacatctt ggacaaggac 1320
ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380ccatacttga gacacttcgc ttacgatcct caatacttct tgaacgagtt ggacttgctt 1380
ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440ggtcaggctg ccgctactca attggctaga aacatctcta actccggtgc catgcaaact 1440
tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500tactttgctg gtgaaaccat tccaggtgac aacttggcct acgatgctga cttgagagct 1500
tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560tgggttgaat acattccata ccacttcaga cctaactacc atggtgtcgg aacctgttct 1560
atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctaacgttta cggtgtccag 1620atgatgccaa aggagatggg tggtgtcgtt gacaacgccg ctaacgttta cggtgtccag 1620
ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680ggattgagag ttatcgacgg ttctatccca cctactcaaa tgtcctctca cgttatgacc 1680
gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740gtcttctacg ctatggcttt gaagatcgca gacgctgttt tggctgacta cgcctccatg 1740
caataa 1746caataa 1746
<210> 20<210> 20
<211> 30<211> 30
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 20<400> 20
ggtattgagg cttccttgtt gactgaccca 30ggtattgagg cttccttgtt gactgaccca 30
<210> 21<210> 21
<211> 30<211> 30
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 21<400> 21
ttgcatggag gcgtagtcag ccaaaacagc 30ttgcatggag gcgtagtcag ccaaaacagc 30
<210> 22<210> 22
<211> 32<211> 32
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 22<400> 22
tggaccagac catctaaggc tcaagttgac tc 32tggaccagac catctaaggc tcaagttgac tc 32
<210> 23<210> 23
<211> 32<211> 32
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 23<400> 23
aacttgagcc ttagatggtc tggtccaagt ac 32aacttgagcc ttagatggtc tggtccaagt ac 32
<210> 24<210> 24
<211> 32<211> 32
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 24<400> 24
tccgctgttg aaaacgatgg tgtcccaact aa 32tccgctgttg aaaacgatgg tgtcccaact aa 32
<210> 25<210> 25
<211> 32<211> 32
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 25<400> 25
tgggacacca tcgttttcaa cagcggacat ca 32tgggacacca tcgttttcaa cagcggacat ca 32
<210> 26<210> 26
<211> 32<211> 32
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 26<400> 26
gactacactg agcaagctca cgaattgctt aa 32gactacactg agcaagctca cgaattgctt aa 32
<210> 27<210> 27
<211> 32<211> 32
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 27<400> 27
caattcgtga gcttgctcag tgtagtcacc ga 32caattcgtga gcttgctcag tgtagtcacc ga 32
<210> 28<210> 28
<211> 32<211> 32
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 28<400> 28
caattcgtga gcttgctcag tgtagtcacc ga 32caattcgtga gcttgctcag tgtagtcacc ga 32
<210> 29<210> 29
<211> 32<211> 32
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 29<400> 29
ccaagcatcc aagttagcat cgtaggccaa gt 32ccaagcatcc aagttagcat cgtaggccaa gt 32
<210> 30<210> 30
<211> 32<211> 32
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 30<400> 30
gacaacgccg ctaacgttta cggtgtccag gg 32gacaacgccg ctaacgttta cggtgtccag gg 32
<210> 31<210> 31
<211> 31<211> 31
<212> DNA<212> DNA
<213> 人工序列(Artificial Sequence)<213> Artificial Sequence
<400> 31<400> 31
gacaccgtaa acgttagcgg cgttgtcaac g 31gacaccgtaa acgttagcgg cgttgtcaac g 31
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101528766A (en) * | 2006-08-04 | 2009-09-09 | 维莱尼姆公司 | Glucanases, nucleic acids encoding them and methods for making and using them |
| CN101629204A (en) * | 2003-03-31 | 2010-01-20 | 诺维信股份有限公司 | Methods for producing biological substances in enzyme-deficient mutants of aspergillus |
| AR101611A1 (en) * | 2014-08-21 | 2016-12-28 | Dsm Ip Assets Bv | GLUCOSE OXIDASES TO IMPROVE THE BAKING |
| US20180186882A1 (en) * | 2014-10-14 | 2018-07-05 | Novartis Ag | Antibody molecules to pd-l1 and uses thereof |
| US20180222982A1 (en) * | 2015-07-29 | 2018-08-09 | Novartis Ag | Combination therapies comprising antibody molecules to pd-1 |
| CN110577939A (en) * | 2018-06-07 | 2019-12-17 | 青岛红樱桃生物技术有限公司 | glucose oxidase mutant with improved heat resistance as well as coding gene and application thereof |
| CN110651048A (en) * | 2017-04-11 | 2020-01-03 | 诺维信公司 | Glucoamylase variants and polynucleotides encoding them |
| CN112301009A (en) * | 2019-07-26 | 2021-02-02 | 中国农业科学院北京畜牧兽医研究所 | Glucose oxidase mutant GOD with improved heat stability as well as gene and application thereof |
| CN113862233A (en) * | 2021-12-03 | 2021-12-31 | 中国农业科学院北京畜牧兽医研究所 | Method for improving acid stability of glucose oxidase, mutant Q241E/R499E, gene and application |
-
2022
- 2022-06-09 CN CN202210648724.1A patent/CN114736880B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101629204A (en) * | 2003-03-31 | 2010-01-20 | 诺维信股份有限公司 | Methods for producing biological substances in enzyme-deficient mutants of aspergillus |
| CN101528766A (en) * | 2006-08-04 | 2009-09-09 | 维莱尼姆公司 | Glucanases, nucleic acids encoding them and methods for making and using them |
| AR101611A1 (en) * | 2014-08-21 | 2016-12-28 | Dsm Ip Assets Bv | GLUCOSE OXIDASES TO IMPROVE THE BAKING |
| US20180186882A1 (en) * | 2014-10-14 | 2018-07-05 | Novartis Ag | Antibody molecules to pd-l1 and uses thereof |
| US20180222982A1 (en) * | 2015-07-29 | 2018-08-09 | Novartis Ag | Combination therapies comprising antibody molecules to pd-1 |
| CN110651048A (en) * | 2017-04-11 | 2020-01-03 | 诺维信公司 | Glucoamylase variants and polynucleotides encoding them |
| CN110577939A (en) * | 2018-06-07 | 2019-12-17 | 青岛红樱桃生物技术有限公司 | glucose oxidase mutant with improved heat resistance as well as coding gene and application thereof |
| CN112301009A (en) * | 2019-07-26 | 2021-02-02 | 中国农业科学院北京畜牧兽医研究所 | Glucose oxidase mutant GOD with improved heat stability as well as gene and application thereof |
| CN113862233A (en) * | 2021-12-03 | 2021-12-31 | 中国农业科学院北京畜牧兽医研究所 | Method for improving acid stability of glucose oxidase, mutant Q241E/R499E, gene and application |
Non-Patent Citations (2)
| Title |
|---|
| 曾庆梅等: "黑曲霉酸性α-淀粉酶基因的克隆及其在毕赤酵母中的表达", 《食品科学》 * |
| 闻一凡等: "定点突变提高毕赤酵母产葡萄糖氧化酶的氧化稳定性", 《食品与生物技术学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023225459A2 (en) | 2022-05-14 | 2023-11-23 | Novozymes A/S | Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections |
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