CN112175919A - Lactone hydrolase mutant and application thereof - Google Patents

Lactone hydrolase mutant and application thereof Download PDF

Info

Publication number
CN112175919A
CN112175919A CN202011062911.9A CN202011062911A CN112175919A CN 112175919 A CN112175919 A CN 112175919A CN 202011062911 A CN202011062911 A CN 202011062911A CN 112175919 A CN112175919 A CN 112175919A
Authority
CN
China
Prior art keywords
val
gly
thr
leu
pro
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202011062911.9A
Other languages
Chinese (zh)
Other versions
CN112175919B (en
Inventor
万南微
陈永正
张秋华
周中平
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jiangxi Xiongdi Pharmaceutical Co ltd
Original Assignee
Jiangxi Xiongdi Pharmaceutical Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Jiangxi Xiongdi Pharmaceutical Co ltd filed Critical Jiangxi Xiongdi Pharmaceutical Co ltd
Priority to CN202011062911.9A priority Critical patent/CN112175919B/en
Publication of CN112175919A publication Critical patent/CN112175919A/en
Application granted granted Critical
Publication of CN112175919B publication Critical patent/CN112175919B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
    • C12P41/003—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by ester formation, lactone formation or the inverse reactions
    • C12P41/005—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by ester formation, lactone formation or the inverse reactions by esterification of carboxylic acid groups in the enantiomers or the inverse reaction
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/42—Hydroxy-carboxylic acids

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Mycology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

The scheme discloses a lactone hydrolase mutant in the fields of genetic engineering technology and biological catalysis, and application thereof in preparing optically pure D-pantoic acid by splitting D, L-pantoic acid lactone as a biocatalyst. The lactone hydrolase mutant is obtained by taking lactohydrolase from Fusarium verticillium as a parent enzyme and performing single mutation or combined mutation on 1-14 amino acid sites in an amino acid sequence SEQ NO. 2; compared with the parent enzyme, the hydrolysis activity of the lactone hydrolase mutant on D, L-pantoic acid lactone is improved by 2.3 to 128 times. The high-activity lactonohydrolase provided by the invention can be used for large-scale production of preparing optical pure D-pantoic acid by biological resolution of D, L-pantoic acid lactone.

Description

Lactone hydrolase mutant and application thereof
Technical Field
The invention belongs to the fields of genetic engineering technology and biocatalysis, and particularly relates to a lactone hydrolase mutant and application thereof.
Background
D-pantoic acid is a key chiral intermediate for the manufacture of calcium D-pantothenate, D-panthenol, D-pantethine. Calcium D-pantothenate is widely used in the feed, pharmaceutical and food industries, and has become a second-order vitamin to niacin, especially as an important feed additive. D-panthenol can be used as a skin remedy for preventing skin infection and is effective in treating skin wounds, skin abrasions and scalds. D-pantethine is used in the pharmaceutical industry as a hypolipidemic agent, and can reduce total cholesterol and triglycerides in serum and increase high density lipoprotein cholesterol in serum.
The preparation method of the D-pantoic acid comprises a chemical resolution method, a physical resolution method and a biological enzyme method resolution method. The chemical resolution technology needs expensive resolution reagents such as quinic acid, strychnine reduction and the like, the separation process is complex, and the pollution problem is serious; the physical resolution technology has harsh conditions, low optical purity and low yield. Therefore, the production process of D-pantoic acid chemical resolution and material resource resolution has higher cost, and the product quality is often difficult to ensure. The biological enzyme method resolution technology uses the lactone hydrolase with high stereoselectivity, the reaction condition is mild, the catalyst is cheap, the environmental pollution is little, the separation and purification process is simple and convenient, the optical purity of the product is high, and the production cost is low. At present, the global production of D-pantoic acid mainly adopts a biological enzyme method resolution technology.
Sakamoto Keiji et al, Kyoto university, Japan, screened a D-pantolactone hydrolase-producing microorganism strain from the environment and identified it as Fusarium oxysporum F.oxysporum. The strain can asymmetrically hydrolyze D-pantoic acid lactone in D, L-pantoic acid lactone with high selectivity to obtain D-pantoic acid. The secondary enzymatic resolution technique has been used for the industrial production of D-pantoic acid by Fuji corporation of Japan (US Patent 5,275,949,1994; US Patent 5,372,940,1994).
The microbial strain producing D-pantolactone hydrolase is screened from the environment by professor sunkenghaiensis of university of south of the Yangtze river and identified as Fusarium moniliforme SW-902 (preservation number is CGMCC No. 0563). The strain can also carry out high-selectivity asymmetric hydrolysis on D-pantoic acid lactone in D, L-pantoic acid lactone to obtain D-pantoic acid. Zhejiang Xin biochemical-rich GmbH company has used the biotechnology for the industrial production of D-pantoic acid, and the enzymatic production of D-pantoic acid is firstly realized at home (CN 1313402A).
The enzyme activities of the two microorganisms reach the requirements of industrial production, but the hydrolysis reaction time is longer and the catalyst dosage is larger in the actual production process. In order to improve the expression quantity of the lactone hydrolase in cells, the construction of recombinant lactone hydrolase gene engineering bacteria by a genetic engineering technology is an effective method. The first precision chemical company of Japan clones the natural lactone hydrolase gene of Fusarium oxysporum F.oxysporum into a cephalosporium apicompleyi or Aspergillus oryzae host by complicated gene recombination expression design, and constructs a corresponding gene engineering bacterium producing lactonohydrolase (WO 2004078951). The Aspergillus oryzae engineering bacteria express lactone hydrolase intracellularly, and the Sporotrichum acremonium secretes lactone hydrolase to the outside of the cell, but the enzyme activity of the two engineering bacteria producing lactone hydrolase gene is not obviously improved, and even the activity of the engineering bacteria is lower than that of wild Fusarium oxysporum. In addition, the above-mentioned microorganisms producing lactonohydrolase are filamentous fungi, and the cells thereof are in the form of various forms of hyphae, and the preparation of immobilized cells (catalysts) by filamentous fungi is difficult and has poor stability as compared with unicellular microorganisms.
Disclosure of Invention
In order to solve the problems, the invention provides a series of lactone hydrolase mutants with improved activity and recombinant genetic engineering bacteria thereof by a genetic engineering technology.
The lactone hydrolase mutant in the scheme is specifically a lactone hydrolase mutant which is obtained by using lactohydrolase (amino acid sequence SEQ NO:2) derived from Fusarium verticillium as a parent enzyme and performing directed evolution on protein to obtain a series of activity-improved lactone hydrolase mutants (amino acid sequence SEQ NO:4-78, wherein the sequence number is an even number) and contains 1-14 amino acid sites and is obtained by single mutation or multi-point combined mutation, and the specific steps are as follows:
the lactone hydrolase mutant is obtained by carrying out single mutation on any one of the following conditions of an amino acid sequence shown as SEQ ID NO: 2:
(1) mutating lysine at position 66 to glutamic acid to obtain mutant with amino acid sequence of SEQ ID NO. 56;
(2) the 147 th lysine is mutated into glutamine to obtain a mutant with the amino acid sequence of SEQ ID NO 60;
(3) the 163 th aspartic acid is mutated into histidine, and the obtained mutant amino acid sequence is SEQ ID NO. 62;
(4) the 163 th aspartic acid is mutated into tyrosine, and the obtained mutant amino acid sequence is SEQ ID NO: 64;
(5) the 341 th tyrosine is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 66;
(6) valine at the 260 th site is mutated into alanine, and the obtained mutant amino acid sequence is SEQ ID NO. 68;
(7) valine at the 260 th position is mutated into leucine, and the obtained mutant amino acid sequence is SEQ ID NO. 70;
(8) mutating the 228 th glycine into valine to obtain a mutant with an amino acid sequence of SEQ ID NO: 72;
(9) the 319 th lysine is mutated into leucine to obtain a mutant with the amino acid sequence of SEQ ID NO: 74;
(10) the 352 nd aspartic acid is mutated into phenylalanine, and the obtained mutant amino acid sequence is SEQ ID NO. 76;
(11) the 356 th lysine is mutated into arginine to obtain the mutant with the amino acid sequence of SEQ ID NO. 78.
The lactone hydrolase mutant is obtained by carrying out multipoint combined mutation on an amino acid sequence shown as SEQ ID NO. 2, specifically, the 66 th lysine is mutated into glutamic acid, the 147 th lysine is mutated into glutamine, the 172 th glycine is mutated into serine, and the 163 th aspartic acid is mutated under any one of the following conditions:
(1) mutating the 163 th aspartic acid into histidine to obtain a mutant with an amino acid sequence of SEQ ID NO. 4;
(2) the 163 th aspartic acid is mutated into tyrosine, and the obtained mutant amino acid sequence is SEQ ID NO 6.
Further, the lactone hydrolase mutant is obtained by mutating the amino acid sequence SEQ ID NO. 6 of the mutant in any one of the following cases:
(1) valine at the 260 th site is mutated into alanine, and the obtained mutant amino acid sequence is SEQ ID NO. 8;
(2) valine at the 260 th site is mutated into leucine, and the obtained mutant amino acid sequence is SEQ ID NO. 10;
(3) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, and the obtained mutant amino acid sequence is SEQ ID NO. 12;
(4) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 14;
(5) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into cysteine, and the obtained mutant amino acid sequence is SEQ ID NO. 16;
(6) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, and aspartic acid at the 226 th site is mutated into asparagine, and the obtained mutant amino acid sequence is SEQ ID NO. 18;
(7) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, and aspartic acid at the 226 th site is mutated into serine, so that the amino acid sequence of the obtained mutant is SEQ ID NO. 20;
(8) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, aspartic acid at the 226 th site is mutated into asparagine, and glutamic acid at the 141 th site is mutated into alanine, and the obtained mutant amino acid sequence is SEQ ID NO. 22;
(9) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, aspartic acid at the 226 th site is mutated into asparagine, glutamic acid at the 141 th site is mutated into alanine, and phenylalanine at the 327 th site is mutated into tyrosine, and the obtained mutant amino acid sequence is SEQ ID NO. 24;
(10) valine at the 260 th position is mutated into alanine, isoleucine at the 88 th position is mutated into phenylalanine, tyrosine at the 341 th position is mutated into serine, aspartic acid at the 226 th position is mutated into asparagine, glutamic acid at the 141 th position is mutated into alanine, phenylalanine at the 327 th position is mutated into tyrosine, threonine at the 4 th position is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 26;
(11) glutamic acid 141 is mutated into alanine, and threonine 4 is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 28.
The lactone hydrolase mutant is a multi-point combined mutation carried out by an amino acid sequence shown as SEQ ID NO. 2, and specifically comprises the following steps: the 147 th lysine is mutated into glutamine, the 141 th glutamic acid is mutated into alanine, the 4 th threonine is mutated into serine, and the 327 th phenylalanine is mutated under any one of the following conditions:
(1) the 327 th phenylalanine is mutated into tyrosine, and the obtained mutant amino acid sequence is SEQ ID NO: 36;
(2) the 327 th phenylalanine is mutated into glycine, and the obtained mutant amino acid sequence is SEQ ID NO 38.
Further, the lactone hydrolase mutant is subjected to mutation in any one of the following cases from the amino acid sequence SEQ ID NO:36 of the mutant:
(1) the 66 th lysine is mutated into glutamic acid, the 172 th glycine is mutated into serine, and the 260 th valine is mutated into alanine, so that the amino acid sequence of the mutant is SEQ ID NO. 30;
(2) the 66 th lysine is mutated into glutamic acid, the 226 th aspartic acid is mutated into asparagine, and the obtained mutant amino acid sequence is SEQ ID NO. 32;
(3) the aspartic acid at the 226 th site is mutated into asparagine, and the obtained mutant amino acid sequence is SEQ ID NO. 34;
(4) the aspartic acid at the 226 th site is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO. 40;
(5) the 66 th lysine is mutated into glutamic acid, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO. 42;
(6) the 88 th isoleucine is mutated into alanine, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO. 44;
(7) the 88 th isoleucine is mutated into phenylalanine, the 163 th asparagine is mutated into tyrosine, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO. 46.
The lactone hydrolase mutant is obtained by carrying out multipoint combined mutation on any one of the following conditions of an amino acid sequence shown as SEQ ID NO: 2:
(1) the 147 th lysine is mutated into glutamine, the 163 th asparagine is mutated into tyrosine, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO: 48;
(2) the 66 th lysine is mutated into glutamic acid, the 147 th lysine is mutated into glutamine, the 163 th asparagine is mutated into tyrosine, the 172 th glycine is mutated into serine, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO 50;
(3) the 147 th lysine is mutated into glutamine, the 163 th asparagine is mutated into tyrosine, the 226 th aspartic acid is mutated into methionine, the 88 th isoleucine is mutated into phenylalanine, and the 341 th tyrosine is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 52.
The lactone hydrolase mutant is a multi-point combined mutation carried out by an amino acid sequence shown as SEQ ID NO. 2, and specifically comprises the following steps: the 66 th lysine is mutated into glutamic acid, the 260 th valine is mutated into alanine, the 88 th isoleucine is mutated into phenylalanine, and the 341 th tyrosine is mutated into serine, so that the obtained mutant amino acid sequence is SEQ ID NO. 54.
The invention also provides an escherichia coli genetic engineering bacterium and a yeast genetic engineering bacterium which respectively construct and produce the lactone hydrolase mutant under any condition by taking escherichia coli or yeast as host cells.
Further, the host cell adopted by the lactone hydrolase escherichia coli genetic engineering bacterium is escherichia coli, preferably E.coli BL21(DE 3); the vector for correspondingly expressing the lactone hydrolase can be pET-21a (+), pET-22a (+), pET-22a (+), pET-28a (+), pETDuet-1, and is preferably pET-28a (+) vector; but are not limited to, the E.coli host and the expression vector.
Further, host cell yeast adopted by the lactone hydrolase yeast genetic engineering bacteria is preferably pichia pastoris GS 115; the shuttle vector corresponding to the expression of the lactone hydrolase may be pPICZA, pPICZB, pPICZC, pPICZ alpha A, pPICZ alpha B, pPICZ alpha C; the pPICZA vector is preferred as a vector for extracellular expression of the lactone hydrolase mutant, but is not limited to the yeast host and shuttle vectors described.
Further, microbial cells or enzyme solution containing the recombinant lactonohydrolase mutant are obtained by fermentation of escherichia coli genetic engineering bacteria and yeast genetic engineering bacteria;
wherein, the culture medium for producing the lactone hydrolase by fermenting the escherichia coli genetically engineered bacteria is LB culture medium (peptone 10g/L, yeast extract 5g/L, sodium chloride 10g/L and pH neutral), 2 XYT culture medium (peptone 16g/L, yeast extract 10g/L, sodium chloride 5g/L and pH neutral), TB culture medium (peptone 12g/L, yeast extract 24g/L, glycerol 5g/L and pH neutral), preferably TB culture medium; the culture temperature is 25-37 ℃, the culture temperature in the early stage of fermentation is preferably 35 ℃, and the culture temperature in the enzyme production stage is preferably 30 ℃.
Fermentation product of yeast gene engineering bacteriaThe lactonohydrolase culture medium is BMGY culture medium (peptone 20g/L, yeast extract 10g/L, glycerol 10g/L, YNB 13.4g/L, 4 × 10-4g/L, pH 6.0-6.5), and the culture temperature is 25-35 deg.C, preferably 30 deg.C.
Further, microbial cells expressing the mutant lactonohydrolase intracellularly are collected by centrifugation, or extracellular secretion-expressed lactonohydrolase mutant enzyme solution is obtained by centrifugation and concentration. The microorganism wet cell or concentrated enzyme solution containing lactone hydrolase mutant can be refrigerated in a refrigerator at-20 deg.C for use; the microbial cells or concentrated enzyme solution containing the lactone hydrolase mutant can also be freeze-dried to obtain freeze-dried cells or freeze-dried enzyme powder for later use.
Furthermore, the D, L-pantoic acid lactone is catalyzed and resolved by taking a microorganism wet cell (or freeze-dried cell) or an enzyme solution (or freeze-dried enzyme powder) containing the lactone hydrolase mutant as a biocatalyst to prepare the optically pure D-pantoic acid.
The wet microbial cell is used as a biocatalyst, and the biotransformation system is as follows: dissolving D, L-pantoic acid lactone with water to prepare a substrate solution with the concentration of 50-350 g/L; adding 100 plus 200mL of substrate solution into a 500mL three-necked flask, and magnetically stirring for 200-300 r/min; hydrolysis was initiated by adding 1-10g of frozen wet cells containing the lactone hydrolase mutant. Controlling the hydrolysis reaction temperature to be 25-35 ℃, and preferably 30 ℃; the reaction pH is detected on line by a pH meter and is controlled between 6.5 and 7.5, preferably 7.0 +/-0.2, by feeding 15% ammonia water. The reaction time is 2-24 hours, and the reaction is stopped when the pH value of the reaction is basically stable. Taking 10mL of reaction solution, and centrifuging to remove microbial cells; taking 1mL of supernatant fluid into a 50mL volumetric flask, and diluting by 50 times with deionized water; 2mL of the diluted liquid film was filtered and analyzed by HPLC.
Taking freeze-dried enzyme powder as a biocatalyst, and a biotransformation system comprises the following components: dissolving D, L-pantoic acid lactone with water to prepare a substrate solution with the concentration of 50-350 g/L; adding 100 plus 200mL of substrate solution into a 500mL three-necked flask, and magnetically stirring for 200-300 r/min; adding 0.1-1g of freeze-dried lactonohydrolase mutant enzyme powder, and starting hydrolysis reaction. Controlling the hydrolysis reaction temperature to be 25-35 ℃, and preferably 30 ℃; the reaction pH is detected on line by a pH meter and is controlled between 6.5 and 7.5, preferably 7.0 +/-0.2, by feeding 15% ammonia water. The reaction time is 2-24 hours, and the reaction is stopped when the pH value of the reaction is basically stable. Taking 10mL of reaction solution, and filtering to remove solid particles; taking 1mL of supernatant fluid into a 50mL volumetric flask, and diluting by 50 times with deionized water; 2mL of the diluted liquid film was filtered and analyzed by HPLC.
The beneficial technical effects of the invention are as follows: compared with the wild type Fusarium verticillium lactone hydrolase (amino acid sequence SEQ ID:2), the lactone hydrolase mutant provided by the invention has the advantage that the activity of resolving D, L-pantolactone is improved by 2.3-128 times. In addition, the genetic engineering of the lactonohydrolase mutant provided by the invention is unicellular microorganism, the fermentation culture technology is mature, the obtained lactone hydrolase catalyst has high activity and good stability, and can be used as a biocatalyst for producing and applying the optically pure D-pantoic acid by dynamically splitting high-concentration D, L-pantoic acid lactone.
Drawings
FIG. 1 is a reaction formula of resolving D, L-pantoic acid lactone to prepare optically pure D-pantoic acid by using a lactone hydrolase mutant as a biocatalyst;
FIG. 2 is an SDS-PAGE identification map of recombinant lactonohydrolase mutants expressed in E.coli hosts: lane M is the standard protein marker; lane 1 is recombinant lactonohydrolase E.coli engineered bacterium; lane 2 is E.coli not carrying the recombinant lactonohydrolase gene.
Detailed Description
A series of lactone hydrolase mutants constructed in the publication have amino acid sequences shown in SEQ ID NO. 4-78 (wherein, the even-numbered sequences in 4-78), and are derived from Fusarium verticillium species such as: 2 and 1, and carrying out single-point mutation or multi-point combined mutation on 1-14 amino acid sites to obtain the novel protein. The mutated protein has the function of efficiently catalyzing the dynamic resolution of D, L-pantoic acid lactone to prepare optically pure D-pantoic acid. The gene sequences corresponding to the amino acid sequences of the lactone hydrolase and its mutants are shown in Table 1 below, wherein it is emphasized that the amino acid sequences corresponding to the mutation of the lactone hydrolase are not limited to those shown in the Table, but also include the gene sequences encoding the same amino acids obtained by "synonymous mutation".
The term "synonymous mutation" means that a nucleotide of a gene is substituted, and the amino acid sequence encoded by the nucleotide sequence after the substitution is identical to the amino acid sequence encoded by the nucleotide sequence before the substitution.
Table 1: lactone hydrolase mutant and corresponding amino acid sequence and coding gene base sequence thereof
Figure BDA0002712877890000071
Figure BDA0002712877890000081
For convenience, the abbreviations for amino acids of proteins may be expressed in either three or one letter English, as is well known to those skilled in the art, and the amino acids corresponding to these abbreviations are shown in Table 2 below:
table 2: amino acid abbreviation and Chinese-English comparison table
A Ala Alanine Alanine
C Cys Cysteine Cysteine
D Asp Aspartic acid Aspartic acid
E Glu Glutamic acid Glutamic acid
F Phe Phenylalanine Phenylalanine
G Gly Glycine Glycine
H His Histidine Histidine
I Ile Isoleucine Isoleucine
K Lys Lysine Lysine
L Leu Leucine Leucine
M Met Methionine Methionine
N Asn Asparagine Asparagine
P Pro Proline Proline
Q Gln Glutamine Glutamine
R Arg Arginine Arginine
S Ser Serine Serine
T Thr Threonine Threonine
V Val Valine Valine
W Trp Tryptophan Tryptophan
Y Tyr Tyrosine Tyrosine
In order to obtain high-activity and high-stability lactone hydrolase for resolving D, L-pantoic acid lactone to prepare high-optical-purity D-pantoic acid lactone, the invention carries out error-prone mutation and point saturation mutation on a gene sequence SEQ ID NO:1 of a code lactone hydrolase SEQ NO:2 to obtain a mutant gene code lactone hydrolase mutation library; further, a series of lactone hydrolase mutants LT 1-LT 38 with improved activities were obtained by activity screening, and the corresponding amino acid sequences and coding gene sequences thereof are shown in Table 1. These lactone hydrolase mutants comprise threonine at position 4, lysine at position 66, isoleucine at position 88, glutamic acid at position 141, lysine at position 147, asparagine at position 163, glycine at position 172, glycine at position 228, valine at position 260, lysine at position 319, asparagine at position 352, phenylalanine at position 327, tyrosine at position 341 and lysine at position 356, and a single mutant or a combined mutant of 14 amino acids in total. Compared with wild lactone hydrolase, the activity of hydrolase D, L-pantolactone of the lactone hydrolase mutants LT 1-LT 38 is improved by 2.3-128 times.
The invention also provides a method for expressing the lactone hydrolase mutants by constructing genetically engineered bacteria. When the amino acid sequence of the lactone hydrolase mutant is known, the genes encoding it, expression cassettes and plasmids containing the genes, and transformants containing the plasmids are readily available to those skilled in the art. These genes, expression cassettes, plasmids and transformants can be constructed by genetic engineering methods well known to those skilled in the art.
The above-mentioned transformant host for expressing the lactone hydrolase-producing mutants may be any microorganism suitable for expressing these mutants, including bacteria and fungi, preferably E.coli and yeast.
When used as biocatalysts for production, the lactone hydrolase mutants of the invention can be in the form of cells or isolated enzymes. The cell forms comprise wet cells, freeze-dried cells, immobilized cells and the like; the form of the separating enzyme comprises crude enzyme liquid, pure enzyme, immobilized enzyme and the like.
The present invention will be described in further detail with reference to specific examples. It should be understood that the following examples are illustrative of the present invention only and are not intended to limit the scope of the present invention.
The addition amount, content and concentration of various substances are referred to herein, wherein the percentage refers to the mass percentage unless otherwise specified.
The molecular biology experiments in the embodiment include experimental techniques such as preparation of host competent cells, construction of plasmids, restriction and ligation of genes, plasmid transformation, PCR gene amplification and the like, and are mainly carried out with reference to molecular cloning experimental guidelines (third edition), and detailed and feasible experimental conditions can be determined through simple preliminary experiments if necessary; the reagents, plasmids and host cells involved in molecular cloning are all obtained from commercial companies.
Example 1: construction of wild-type lactone hydrolase-expressing Strain
For the derived lactone hydrolase LT0, according to the reported coding gene sequence SEQ ID NO:1, the Wuhan Kirginia bioengineering Co., Ltd is entrusted to complete gene sequence synthesis, subcloned to pGM-T vector, transformed to Escherichia coli E.coli DH5 alpha host, and the recombinant plasmid pGM-T-LT0 transformant containing the lactone hydrolase gene is obtained.
Designing a cloning primer:
forward primer F1: 5' -CATATGCCTTCCACCATTTCTG-3’
Forward primer R1: 5' -CTCGAGCTAGTCATACAACTTG-3’
pGM-T-LT0 plasmid was extracted as a gene template, and LT 0-encoding gene was amplified by PCR using F1 and R1 forward and reverse primers to amplify a fragment of about 1.2 kb. 50 microliter of PCR reaction system: 1 microliter each of primers F1 and R1, 3 microliter plasmid template, 25 microliter 2X Premix PrimeSTAR HS, 10 microliter sterile distilled water. PCR procedure: 5min at 95 ℃; 10s at 98 ℃, 5s at 55 ℃ and 1.5min at 72 ℃ for 30 cycles; finally 72 ℃ for 10 min.
After the PCR amplification is finished, 5 microliters of PCR products are detected by agarose gel electrophoresis, and after the 1.2kb fragment is determined to be amplified, the residual PCR products are recovered into the target gene fragment by a gel recovery kit. The amplified target gene has NdeI restriction site at 5 'end and XhoI restriction site at 3' end.
Further, the recovered lactone hydrolase gene fragment was digested with NdeI and XhoI, and pET-28a (+) plasmid vector was simultaneously digested. And recovering the digested LT0 gene fragment and the large vector fragment by using a gel recovery kit.
Further, ligation was performed overnight at 16 ℃ in a metal bath by T4 DNA ligase. Gene ligation system 20 microliters: 14 microliter of vector fragment, 5 microliter of LT0 gene fragment and 1 microliter of T4 DNA ligase.
Further, 10 microbial overnight ligations were taken to transform E.coli DH 5. alpha. competent host cells, plated on solid LB plates containing kanamycin sulfate. The components of the culture medium are as follows: 10g/L of peptone, 5g/L of yeast extract, 10g/L of sodium chloride, 15g/L of agar powder and 50 mu g/mL of kanamycin sulfate. And (3) inversely placing the plate containing the transformation liquid in a constant-temperature incubator at 37 ℃ for culturing for 12-16 hours, and growing an escherichia coli colony on a solid LB culture medium.
Furthermore, after colony PCR verification, a plurality of recombinant clones are picked out for culture, and Shanghai biological Limited company is entrusted to carry out sequencing analysis on recombinant plasmid pET-28-LT0 genes in the host, so as to determine that the target genes in the plasmids contained in the host cells are completely correct.
Further, the pET-28-LT0 plasmid in e.coli DH5 α host was extracted by a plasmid extraction kit, transformed into expression host cell e.coli BL21(DE3) competent, and plated on a solid LB plate containing kanamycin sulfate. The components of the culture medium are as follows: 10g/L of peptone, 5g/L of yeast extract, 10g/L of sodium chloride, 15g/L of agar powder and 50 mu g/mL of kanamycin sulfate. And (3) inversely placing the plate containing the transformation liquid in a constant-temperature incubator at 37 ℃ for culturing for 12-16 hours, and growing an escherichia coli colony on a solid LB culture medium. Through colony PCR verification, the Escherichia coli genetic engineering bacteria capable of expressing the lactone hydrolase LT0 are obtained.
Fermentation of intracellular products: and transferring the escherichia coli genetic engineering bacteria into a 250mL shake bottle for culture, wherein the shake bottle contains 50mL of liquid LB culture medium. The components of the culture medium are as follows: 10g/L of peptone, 5g/L of yeast extract, 10g/L of sodium chloride and 50 mu g/mL of kanamycin sulfate. Carrying out shake culture at the constant temperature of 37 ℃, adding IPTG with the final concentration of 0.5mM when the OD600 value of the cell concentration is 0.6-1.0, inducing the expression of the recombinant lactonohydrolase LT0 in the cell, and inducing the continuous growth for 12-14 hours at the temperature of 37 ℃. After fermentation is finished, centrifuging the fermentation liquor, collecting microbial cells containing the lactone hydrolase mutant, and storing the microbial cells in a refrigerator at the temperature of 20 ℃ below zero for later use; or freeze drying the microbial cells in one step to obtain lyophilized cells, and storing in a refrigerator at-20 deg.C.
Example 2: obtaining lactone hydrolase mutant with excellent performance by directed evolution
Designing a mutation primer:
forward primer F2: 5'-TAATACGACTCACTATAGGG-3'
Reverse primer R2: 5'-TCGCTTGGAACATTATTAAC-3'
A random mutation library is constructed by an error-prone PCR technology by taking pET-28-LT0 plasmid containing a gene sequence of SEQ ID NO. 1 as a template and F2 and R2 as positive and negative primers. Error-prone PCR system 50 μ l: 2 XTaqPCR MaterMix 25. mu.l, primers F2 and R2 each 1. mu.l, pET-28-LT0 plasmid 1. mu.l, dGTP (10mM) 0.5. mu.l, MnCl 21 microliter (10mM) and 20.5 microliter of sterile water. PCR conditions were as follows:5min at 95 ℃; 30 cycles of 95 ℃ for 30s, 55 ℃ for 30s and 72 ℃ for 1.5 min; finally 72 ℃ for 10 min.
And carrying out agarose gel electrophoresis detection on the PCR product, wherein the amplification product is about 1200 bp. And recovering an error-prone PCR gene product by using a gel recovery kit, and performing whole plasmid amplification by using KOD-Plus-Neo high-assurance DNA polymerase by using the error-prone PCR gene product as a large primer. Large primer PCR system 50 μ l: 10 x buffer 5. mu.l, KOD-Plus-Neo polymerase 2.5. mu.l, error-prone PCR gene product 8. mu.l, pET-28-LT0 plasmid 0.5. mu.l, dNTPmix 5. mu.l, MgSO43 microliters (25mM) and 26 microliters of sterile water. Large primer PCR procedure: 5min at 68 ℃; 2min at 98 ℃; 30 cycles of 10s at 98 ℃ and 7min at 68 ℃; finally, 68 ℃ for 10 min.
After the large primer PCR was completed, 2.0. mu.l of NEB DpnI restriction enzyme was added, and the methylated template plasmid was digested by incubation at 37 ℃ for 3 hours. Carrying out digestion PCR at 65 ℃ for 30min, taking 10 microliters of the PCR solution, transforming the PCR solution to an expression host cell E.coli BL21(DE3) competence, and carrying out transformation for 5 times; the transformation solution was plated on plates containing screening solid medium at 100. mu.l/plate. Screening the culture medium components: 10g/L of peptone, 5g/L of yeast extract, 10g/L of sodium chloride, 15g/L of agar powder, 50 mu g/mL of kanamycin sulfate, 2g/L of D, L-pantolactone and trace bromothymol blue. And (3) inversely placing the plate containing the transformation liquid in a constant-temperature incubator at 37 ℃ for culturing for 16-24 hours, and growing an escherichia coli colony on a solid LB culture medium. And (4) observing through a discoloration ring around the bacterial colony, selecting the bacterial colony which generates a yellow discoloration ring and is obvious, and further verifying the activity and the selectivity.
The wild LT0 colibacillus engineering bacteria and the colony with obvious color changing ring are inoculated into a test tube containing 10mL liquid LB culture medium and cultured for 12 hours at 37 ℃ to be used as seed liquid. Further, respectively inoculating the seed liquid into 500mL shake flasks according to the volume ratio of 2%, placing the shake flasks containing 100mL liquid LB culture medium into a constant-temperature shaking table at 37 ℃ for culture, adding IPTG (isopropyl-beta-D-thiogalactoside) with the final concentration of 0.5mM when the OD600 value of the cell concentration is 0.6-1.0, inducing the recombinant lactonohydrolase mutant to express in the cells, and inducing the recombinant lactonohydrolase mutant to continue to grow for 12-14 hours at 37 ℃. And centrifuging the fermentation liquor, removing supernatant, and collecting microbial cells containing the lactone hydrolase. The wet cells are used as biocatalysts to carry out the D, L-pantoic acid lactone resolution experiment.
Activity verification the biotransformation system was as follows: 20mL of 200mM phosphate buffer pH7.0 was added with 1g D, L-pantolactone, and 0.1g of wet cells, and reacted at 30 ℃ for 10 minutes. After the reaction, 2mL of the reaction solution was centrifuged, the supernatant sample was diluted 10 times, and the diluted solution was subjected to membrane filtration and HPLC analysis.
The conversion was calculated from the concentrations of Pantolactone and (PL) and Pantoic Acid (PA) in the reaction solution. The reaction rate is calculated by the formula: conversion (%) ═ PA concentration/(PL concentration + PA concentration) × 100. The HPLC analysis conditions were as follows:
the optical purity of D-pantoic acid reflects the stereoselectivity of lactone hydrolase, and the calculation formula is as follows: optical purity (%) - (D-PA concentration-L-PA concentration)/(D-PA concentration + L-PA concentration) × 100. The chiral HPLC analytical conditions were as follows: a detector: ultraviolet absorption photometer (measuring wavelength: 254nm) column: lactone 4.6mm, length 50mm, packing diameter 3um, stainless steel tube-silica gel packing (MCI GEL CRS10W) detection temperature: about 30 ℃. Mobile phase: 0.32g of copper sulphate is dissolved in 1000ml of water, 900ml of copper sulphate solution and 100ml of chromatographic acetonitrile are taken. Retention time: the retention time of D-PA was about 4.9 minutes, and the overall retention time was 3.5 minutes for L-PA.
The mutant strain with improved activity was preserved, and the lactone hydrolase gene in the superior mutant strain was subjected to sequencing analysis by Shanghai Biometrics Ltd to determine the amino acid sequence of the superior mutant. Table 3 below is a partial screening result:
table 3 partial screening results:
Figure BDA0002712877890000121
in the next round of mutation screening experiments, the random mutation or the site-specific saturation mutation can be performed on the basis of the excellent lactone hydrolase mutant, namely, the excellent mutant lactone hydrolase mutant gene is used as a template. The point saturation mutation amino acid site can be a hot spot discovered by random mutation, and a potential mutation hot spot selected by computer analysis, and the experimental design concept is well known to those skilled in the art. Further lactone hydrolase mutants with continuously improved activities can be obtained through multiple rounds of random mutation and point saturation mutation. Specifically, the present invention was subjected to 38 rounds of mutation in total, and the mutants having improved activities and the amino acid sequences thereof are shown in Table 1.
Example 3: construction of yeast engineering bacteria and secretion expression of lactonohydrolase mutant
And (3) constructing a yeast engineering bacterium for producing the lactone hydrolase mutant by taking pPICZ alpha A as a shuttle vector and pichia pastoris as an expression host GS115 to realize extracellular secretory expression of the lactone hydrolase mutant.
Designing a cloning primer:
forward primer F3: 5' -GAATTCGCCTTCCACCATTTCTG-3’
Forward primer R3: 5' -GCGGCCGCCTAGTCATACAACTTG-3’
The recombinant plasmid contained in the mutant strain TB4 of the lactone hydrolase obtained in example 2 was pET-28-LT-23, the amino acid sequence of the expressed mutant strain of the lactone hydrolase was SEQ ID NO. 48, and the corresponding gene sequence was SEQ ID NO. 47.
A plasmid pET-28-LT-23 of the mutant strain TB4 is extracted by a plasmid extraction kit, the plasmid is taken as a template gene, F3 and R3 are taken as forward and reverse primers, the lactone hydrolase mutant gene SEQ ID NO:47 is amplified by PCR, and the amplified fragment is about 1.2 kb. 50 microliter of PCR reaction system: 1 microliter each of primers F3 and R3, 3 microliter plasmid template, 25 microliter 2X Premix PrimeSTAR HS, 10 microliter sterile distilled water. PCR procedure: 5min at 95 ℃; 10s at 98 ℃, 5s at 55 ℃ and 1.5min at 72 ℃ for 30 cycles; finally 72 ℃ for 10 min.
After the PCR amplification is finished, 5 microliters of PCR products are detected by agarose gel electrophoresis, and after the 1.2kb fragment is determined to be amplified, the residual PCR products are recovered into the target gene fragment by a gel recovery kit. The 5 'end of the amplified target gene is provided with an EcoRI enzyme cutting site, and the 3' end is provided with a NotI enzyme cutting site.
Further, the recovered lactone hydrolase gene fragment was digested with EcoRI and NotI, and the pPICZ. alpha.A plasmid vector was simultaneously digested. The pPICZ alpha A plasmid vector is extracted from Escherichia coli cloning host bacteria preserved in a laboratory, and the plasmid is a bleomycin resistance plasmid. After the enzyme digestion is finished, the digested LT23 gene fragment and the large vector fragment are recovered by a gel recovery kit.
Further, ligation was performed overnight at 16 ℃ in a metal bath by T4 DNA ligase. Gene ligation system 20 microliters: 16 microliter of vector fragment, 3 microliter of LT23 gene fragment and 1 microliter of T4 DNA ligase.
Further, 10 microbial overnight ligations were taken to transform E.coli DH 5. alpha. competent host cells, plated on solid LB plates containing kanamycin sulfate. The components of the culture medium are as follows: 10g/L of peptone, 5g/L of yeast extract, 10g/L of sodium chloride, 15g/L of agar powder and 25 mu g/mL of bleomycin. And (3) inversely placing the plate containing the transformation liquid in a constant-temperature incubator at 37 ℃ for culturing for 12-16 hours, and growing an escherichia coli colony on a solid LB culture medium.
Furthermore, after colony PCR verification, a plurality of recombinant clones are picked out for culture, and Shanghai biological Limited company is entrusted to carry out sequencing analysis on the recombinant plasmid pPICZ alpha-LT 23 gene in the host, so as to determine that the target gene in the plasmid contained in the host cell is completely correct.
Further, a plasmid extraction kit is used for extracting pPICZ alpha-LT 23 plasmid from E.coli DH5 alpha host, and SacI restriction endonuclease is used for digesting the plasmid to obtain linearized gene. 20 microliter of enzyme digestion system: 1 microliter of SacI endonuclease, 10 microliter of buffer2 microliter, 8 microliter of pPICZ alpha-LT 23 plasmid is extracted, and 9 microliter of sterile water is extracted; the reaction was incubated at 37 ℃ for 1 h. After the enzyme digestion is finished, the enzyme digestion product is recovered by a gel recovery kit, and the size of the recovered gene fragment is about 4.8kb by agarose gel electrophoresis detection.
The linearized plasmid pPICZ alpha-LT 23 fragment recovered from 3. mu.l of gum was transformed into Pichia pastoris GS115 competent. The conditions for electric shock conversion are as follows: voltage 1.5kV, capacitance 25uF, shock time 5 mLsec. Immediately after the shock was completed, 1.0mL of pre-cooled sterile 1mL sorbitol was added and gently mixed. And the transformation solution was rapidly transferred from the electric rotor to a 2.0mL EP tube and incubated at 30 ℃ for 1 hour. And spreading 100 microliter resuscitation transformation liquid on YPD plate containing bleomycin, and culturing in 30 deg.C incubator in dark for 72-96 hr until single colony appears. YPD medium composition: 20g/L of peptone, 10g/L of yeast extract, 20g/L of glucose, 15g/L of agar powder and 10 mu g/mL of bleomycin. Further, after colony PCR verification and SDS-PAGE verification of fermentation liquor, the yeast gene engineering bacteria secreting and expressing the lactone hydrolase mutant LT23 are obtained.
And (3) fermentation extracellular secretion enzyme production: and transferring the saccharomyces cerevisiae engineering bacteria into a 250mL shake bottle for culture, wherein the shake bottle contains 50mL of liquid BMGY culture medium. The components of the culture medium are as follows: peptone 20g/L, yeast extract 10g/L, and glucose 20 g/L. Shaking-culturing at 30 deg.C for 24 hr, centrifuging, collecting thallus, and removing supernatant. Further, after the thalli were resuspended in 10mL of BMMY media, the thalli were transferred to a 500mL shake flask containing 190mL of BMMY media. The culture medium is continuously subjected to shake culture at the constant temperature of 30 ℃ for 72 hours, and 200 microliters of methanol is supplemented every 24 hours for induction. After fermentation is finished, centrifuging the fermentation liquor, and collecting supernatant containing the lactone hydrolase mutant protein; further, concentrating the fermentation liquor by membrane filtration equipment to obtain concentrated enzyme solution, and storing in a refrigerator at-20 deg.C for use; or further freeze drying the concentrated supernatant to obtain lyophilized enzyme powder, and storing in a refrigerator at-20 deg.C. BMGY media components: peptone 20g/L, yeast extract 10g/L, glycerol 10g/L, YNB 13.4g/L, 4X 10-4g/L pH 6.0. BMMY media components: peptone 20g/L, yeast extract 10g/L, methanol 0.5%, YNB 13.4g/L, 4X 10%-4g/L pH 6.0。
Example 4: method for dynamic resolution of D, L-pantolactone by catalyzing resting cells of escherichia coli engineering bacteria
Weighing 10g of wet cells of the engineering bacteria LT3 of escherichia coli collected by fermentation, completely putting the wet cells into 100mL of aqueous solution containing 220g/L lactone for hydrolysis experiment, wherein the hydrolysis condition is that the temperature is 28 ℃, stirring is carried out at 200rpm, 2% ammonia water is adopted in the hydrolysis process to control the pH value to be 6.8-7.0, 1mL of hydrolysate is sampled every 1 hour, 0.22 mu m membrane is adopted to filter filtrate, HPLC chromatographic analysis is carried out, after 8 hours of hydrolysis, the hydrolysis rate is 28.1%, and the optical purity is 90.7% (the kinetic resolution data is shown in Table 4).
TABLE 4 procedure of catalytic reaction of resting cells of engineering bacteria of Escherichia coli
Figure BDA0002712877890000141
Example 5: dynamic resolution of D, L-pantolactone by catalysis of resting cells of yeast engineering bacteria
Weighing 2g of yeast engineering bacteria LT5 wet cells collected by fermentation, carrying out hydrolysis experiment in an aqueous solution containing 220g/L lactone, wherein the hydrolysis condition is that the temperature is 28 ℃, stirring is carried out at 200rpm, 2% ammonia water is adopted in the hydrolysis process, the pH is controlled to be 6.8-7.0, 1mL of hydrolysate is sampled every 0.5 hour, a 0.22 mu m membrane is adopted to filter filtrate, HPLC chromatographic analysis is carried out, the reaction is carried out for 2 hours, the hydrolysis rate of D, L-pantoic acid lactone is 48.2%, and the optical purity of the product D-pantoic acid is 96.7% (the kinetic resolution data is shown in Table 5).
TABLE 5 Yeast engineering bacteria resting cell catalytic reaction course
Figure BDA0002712877890000151
Sequence listing
<110> Jiangxi Brother medicine Co Ltd
<120> lactone hydrolase mutants and uses thereof
<160> 78
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 2
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 2
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 3
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatccacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 4
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 4
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile His Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 5
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 5
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 6
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 6
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 7
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 7
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 8
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 8
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 9
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 9
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccctc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 10
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 10
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Leu Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 11
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 11
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct cttcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 12
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 12
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 13
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 13
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct cttcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tctgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 14
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 14
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Ser Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 15
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 15
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct cttcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tgtgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 16
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 16
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Cys Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 17
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 17
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct cttcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtaactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tctgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 18
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 18
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asn Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Ser Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 19
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 19
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct cttcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtagctt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tctgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 20
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 20
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Ser Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Ser Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 21
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 21
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct cttcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
ggggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtagctt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tctgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 22
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 22
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Ser Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Ser Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 23
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 23
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct cttcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
ggggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtagctt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctta catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tctgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 24
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 24
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Ser Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Tyr Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Ser Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 25
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 25
atgccttcct ctatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct cttcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
ggggtccgca agggcaagca ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagagcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtagctt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctta catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tctgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 26
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 26
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Ser Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Tyr Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Ser Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 27
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 27
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctgaacttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagtcaaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 28
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 28
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 29
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 29
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctgaacttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagtcaaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctta catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 30
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 30
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Tyr Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 31
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 31
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctgaacttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggttcatt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctgg catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 32
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 32
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Ser Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Gly Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 33
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 33
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggttcatt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctta catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 34
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 34
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Ser Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Tyr Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 35
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 35
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttc aatccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 36
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 36
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Ser Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 37
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 37
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctgg catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 38
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 38
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Gly Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 39
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 39
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtatgtt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctta catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 40
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 40
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Met Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Tyr Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 41
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 41
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggaacc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtatgtt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctta catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 42
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 42
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Met Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Tyr Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 43
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 43
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct ctttgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtatgtt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctta catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 44
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 44
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Met Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Tyr Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 45
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 45
atgccttcct caatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct ctttgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gccgtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtatgtt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatctta catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 46
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 46
Met Pro Ser Ser Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Ala Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Met Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Tyr Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 47
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 47
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtatgtt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 48
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 48
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Met Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 49
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 49
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggaacc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagtcaaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtatgtt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 50
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 50
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Asn Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Ser Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Met Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 51
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 51
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct ctttgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtatgtt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tctgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 52
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 52
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Met Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Ser Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 53
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 53
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggaacc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct ctttgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tctgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 54
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 54
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Ser Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 55
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 55
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgaggaacc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 56
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 56
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Glu Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 57
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 57
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct ctttgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 58
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 58
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Phe Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 59
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 59
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagca agacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 60
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 60
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Gln Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 61
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 61
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatccacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 62
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 62
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile His Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 63
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 63
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatctacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 64
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 64
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Tyr Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 65
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 65
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tctgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 66
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 66
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Ser Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 67
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 67
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgcc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 68
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 68
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Ala Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 69
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 69
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccctc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 70
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 70
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Leu Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 71
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 71
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tgttatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 72
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 72
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 73
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 73
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcctgacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 74
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 74
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Leu Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 75
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 75
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggttcccct ctggcaagct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 76
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 76
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Phe
340 345 350
Pro Ser Gly Lys Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400
<210> 77
<211> 1203
<212> DNA/RNA
<213> Artificial Sequence (Artificial Sequence)
<400> 77
atgccttcca ccatttctgt acttgcgggg gttctcgtgc ccgttttggg cgccgtggct 60
gctaagcttc cttctacggc tcagattatc gaccagaagt cgttcaatgt cttgaaggat 120
gtgccgcctc ctgcagtggc caatgactct ctggtgttta cttggcctgg tgtaactgag 180
gagtctttgg tcgagaagcc tttccatgtc tacgatgaag agttttacga cgtcattgga 240
aaggacccct cactgaccct catcgcgaca tcggataccg acccaatttt ccacgaggcc 300
gttgtctggt atcctcctac cgaagaggtc ttctttgtcc agaatgcagg cgctcctgct 360
gctggcactg gattgaacaa gtcttccatc attcagaaga tttccctcaa ggaggccgac 420
gaggtccgca agggcaagaa ggacgaggtc aaggtcaccg tcgttgactc aaaccctcaa 480
gtcatcaacc ccaatggtgg tacttactac aagggcaaca tcatcttcgc tggtgaaggc 540
caaggtgacg atgttccctc cgccctgtac ctgatgaacc ctctccctcc ttacaacact 600
accaccctcc tcaacaacta cttcggtcgc cagttcaact ccctcaacga tgttggcatc 660
aaccccagga atggtgactt gtacttcacc gacaccctct acggatatct ccaagacttc 720
cgccctgttc ctggtctgcg aaaccaagtc tatcgttaca actttgacac cggcgccgtc 780
actgttgtcg ctgatgactt taccctaccc aacggtattg gcttcggtcc cgatggcaag 840
aaggtctatg tcaccgacac tggtatcgct ctcggtttct acggccgcaa cctctcttca 900
cccgcctctg tttactcttt cgacgtgaac caggacggta cactccagaa ccgcaagacc 960
tttgcttacg tcgcatcttt catccccgat ggtgttcata ccgattccaa gggccgtgtt 1020
tatgccggtt gcggcgatgg tgtccatgtc tggaacccct ctggccggct catcggcaag 1080
atctataccg gcactgttgc tgctaacttc cagtttgctg gtaagggaag gatgatcatt 1140
actggacaga caaagctgtt ctatgtgact ttgggagcct caggccccaa gttgtatgac 1200
tag 1203
<210> 78
<211> 400
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 78
Met Pro Ser Thr Ile Ser Val Leu Ala Gly Val Leu Val Pro Val Leu
1 5 10 15
Gly Ala Val Ala Ala Lys Leu Pro Ser Thr Ala Gln Ile Ile Asp Gln
20 25 30
Lys Ser Phe Asn Val Leu Lys Asp Val Pro Pro Pro Ala Val Ala Asn
35 40 45
Asp Ser Leu Val Phe Thr Trp Pro Gly Val Thr Glu Glu Ser Leu Val
50 55 60
Glu Lys Pro Phe His Val Tyr Asp Glu Glu Phe Tyr Asp Val Ile Gly
65 70 75 80
Lys Asp Pro Ser Leu Thr Leu Ile Ala Thr Ser Asp Thr Asp Pro Ile
85 90 95
Phe His Glu Ala Val Val Trp Tyr Pro Pro Thr Glu Glu Val Phe Phe
100 105 110
Val Gln Asn Ala Gly Ala Pro Ala Ala Gly Thr Gly Leu Asn Lys Ser
115 120 125
Ser Ile Ile Gln Lys Ile Ser Leu Lys Glu Ala Asp Glu Val Arg Lys
130 135 140
Gly Lys Lys Asp Glu Val Lys Val Thr Val Val Asp Ser Asn Pro Gln
145 150 155 160
Val Ile Asn Pro Asn Gly Gly Thr Tyr Tyr Lys Gly Asn Ile Ile Phe
165 170 175
Ala Gly Glu Gly Gln Gly Asp Asp Val Pro Ser Ala Leu Tyr Leu Met
180 185 190
Asn Pro Leu Pro Pro Tyr Asn Thr Thr Thr Leu Leu Asn Asn Tyr Phe
195 200 205
Gly Arg Gln Phe Asn Ser Leu Asn Asp Val Gly Ile Asn Pro Arg Asn
210 215 220
Gly Asp Leu Tyr Phe Thr Asp Thr Leu Tyr Gly Tyr Leu Gln Asp Phe
225 230 235 240
Arg Pro Val Pro Gly Leu Arg Asn Gln Val Tyr Arg Tyr Asn Phe Asp
245 250 255
Thr Gly Ala Val Thr Val Val Ala Asp Asp Phe Thr Leu Pro Asn Gly
260 265 270
Ile Gly Phe Gly Pro Asp Gly Lys Lys Val Tyr Val Thr Asp Thr Gly
275 280 285
Ile Ala Leu Gly Phe Tyr Gly Arg Asn Leu Ser Ser Pro Ala Ser Val
290 295 300
Tyr Ser Phe Asp Val Asn Gln Asp Gly Thr Leu Gln Asn Arg Lys Thr
305 310 315 320
Phe Ala Tyr Val Ala Ser Phe Ile Pro Asp Gly Val His Thr Asp Ser
325 330 335
Lys Gly Arg Val Tyr Ala Gly Cys Gly Asp Gly Val His Val Trp Asn
340 345 350
Pro Ser Gly Arg Leu Ile Gly Lys Ile Tyr Thr Gly Thr Val Ala Ala
355 360 365
Asn Phe Gln Phe Ala Gly Lys Gly Arg Met Ile Ile Thr Gly Gln Thr
370 375 380
Lys Leu Phe Tyr Val Thr Leu Gly Ala Ser Gly Pro Lys Leu Tyr Asp
385 390 395 400

Claims (10)

1. A lactone hydrolase mutant characterized by: it is obtained by carrying out single mutation on any one of the following conditions of an amino acid sequence shown as SEQ ID NO: 2:
(1) mutating lysine at position 66 to glutamic acid to obtain mutant with amino acid sequence of SEQ ID NO. 56;
(2) the 147 th lysine is mutated into glutamine to obtain a mutant with the amino acid sequence of SEQ ID NO 60;
(3) the 163 th aspartic acid is mutated into histidine, and the obtained mutant amino acid sequence is SEQ ID NO. 62;
(4) the 163 th aspartic acid is mutated into tyrosine, and the obtained mutant amino acid sequence is SEQ ID NO: 64;
(5) the 341 th tyrosine is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 66;
(6) valine at the 260 th site is mutated into alanine, and the obtained mutant amino acid sequence is SEQ ID NO. 68;
(7) valine at the 260 th position is mutated into leucine, and the obtained mutant amino acid sequence is SEQ ID NO. 70;
(8) mutating the 228 th glycine into valine to obtain a mutant with an amino acid sequence of SEQ ID NO: 72;
(9) the 319 th lysine is mutated into leucine to obtain a mutant with the amino acid sequence of SEQ ID NO: 74;
(10) the 352 nd aspartic acid is mutated into phenylalanine, and the obtained mutant amino acid sequence is SEQ ID NO. 76;
(11) the 356 th lysine is mutated into arginine to obtain the mutant with the amino acid sequence of SEQ ID NO. 78.
2. A lactone hydrolase mutant characterized by: the amino acid sequence is shown as SEQ ID NO. 2 to carry out multipoint combined mutation, specifically, the 66 th lysine is mutated into glutamic acid, the 147 th lysine is mutated into glutamine, the 172 th glycine is mutated into serine, and the 163 th aspartic acid is mutated under any one of the following conditions:
(1) mutating the 163 th aspartic acid into histidine to obtain a mutant with an amino acid sequence of SEQ ID NO. 4;
(2) the 163 th aspartic acid is mutated into tyrosine, and the obtained mutant amino acid sequence is SEQ ID NO 6.
3. A lactone hydrolase mutant characterized by: the mutant of claim 2 having the amino acid sequence SEQ ID NO 6 mutated in any one of the following ways:
(1) valine at the 260 th site is mutated into alanine, and the obtained mutant amino acid sequence is SEQ ID NO. 8;
(2) valine at the 260 th site is mutated into leucine, and the obtained mutant amino acid sequence is SEQ ID NO. 10;
(3) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, and the obtained mutant amino acid sequence is SEQ ID NO. 12;
(4) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 14;
(5) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into cysteine, and the obtained mutant amino acid sequence is SEQ ID NO. 16;
(6) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, and aspartic acid at the 226 th site is mutated into asparagine, and the obtained mutant amino acid sequence is SEQ ID NO. 18;
(7) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, and aspartic acid at the 226 th site is mutated into serine, so that the amino acid sequence of the obtained mutant is SEQ ID NO. 20;
(8) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, aspartic acid at the 226 th site is mutated into asparagine, and glutamic acid at the 141 th site is mutated into alanine, and the obtained mutant amino acid sequence is SEQ ID NO. 22;
(9) valine at the 260 th site is mutated into alanine, isoleucine at the 88 th site is mutated into phenylalanine, tyrosine at the 341 th site is mutated into serine, aspartic acid at the 226 th site is mutated into asparagine, glutamic acid at the 141 th site is mutated into alanine, and phenylalanine at the 327 th site is mutated into tyrosine, and the obtained mutant amino acid sequence is SEQ ID NO. 24;
(10) valine at the 260 th position is mutated into alanine, isoleucine at the 88 th position is mutated into phenylalanine, tyrosine at the 341 th position is mutated into serine, aspartic acid at the 226 th position is mutated into asparagine, glutamic acid at the 141 th position is mutated into alanine, phenylalanine at the 327 th position is mutated into tyrosine, threonine at the 4 th position is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 26;
(11) glutamic acid 141 is mutated into alanine, and threonine 4 is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 28.
4. A lactone hydrolase mutant characterized by: the amino acid sequence is shown as SEQ ID NO 2 to carry out multipoint combined mutation, specifically, the 147 th lysine is mutated into glutamine, the 141 th glutamic acid is mutated into alanine, the 4 th threonine is mutated into serine, and the 327 th phenylalanine is mutated under any one of the following conditions:
(1) the 327 th phenylalanine is mutated into tyrosine, and the obtained mutant amino acid sequence is SEQ ID NO: 36;
(2) the 327 th phenylalanine is mutated into glycine, and the obtained mutant amino acid sequence is SEQ ID NO 38.
5. A lactone hydrolase mutant characterized by: the mutant of claim 4, wherein the amino acid sequence is SEQ ID NO:36, and wherein the mutation is any one of the following:
(1) the 66 th lysine is mutated into glutamic acid, the 172 th glycine is mutated into serine, and the 260 th valine is mutated into alanine, so that the amino acid sequence of the mutant is SEQ ID NO. 30;
(2) the 66 th lysine is mutated into glutamic acid, the 226 th aspartic acid is mutated into asparagine, and the obtained mutant amino acid sequence is SEQ ID NO. 32;
(3) the aspartic acid at the 226 th site is mutated into asparagine, and the obtained mutant amino acid sequence is SEQ ID NO. 34;
(4) the aspartic acid at the 226 th site is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO. 40;
(5) the 66 th lysine is mutated into glutamic acid, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO. 42;
(6) the 88 th isoleucine is mutated into alanine, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO. 44;
(7) the 88 th isoleucine is mutated into phenylalanine, the 163 th asparagine is mutated into tyrosine, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO. 46.
6. A lactone hydrolase mutant characterized by: it is obtained by carrying out multipoint combined mutation on any one of the following conditions of an amino acid sequence shown as SEQ ID NO. 2:
(1) the 147 th lysine is mutated into glutamine, the 163 th asparagine is mutated into tyrosine, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO: 48;
(2) the 66 th lysine is mutated into glutamic acid, the 147 th lysine is mutated into glutamine, the 163 th asparagine is mutated into tyrosine, the 172 th glycine is mutated into serine, and the 226 th aspartic acid is mutated into methionine, and the obtained mutant amino acid sequence is SEQ ID NO 50;
(3) the 147 th lysine is mutated into glutamine, the 163 th asparagine is mutated into tyrosine, the 226 th aspartic acid is mutated into methionine, the 88 th isoleucine is mutated into phenylalanine, and the 341 th tyrosine is mutated into serine, and the obtained mutant amino acid sequence is SEQ ID NO. 52.
7. A lactone hydrolase mutant characterized by: the amino acid sequence of the mutant is shown as SEQ ID NO. 2 to carry out multipoint combined mutation, specifically, the 66 th lysine is mutated into glutamic acid, the 260 th valine is mutated into alanine, the 88 th isoleucine is mutated into phenylalanine, and the 341 th tyrosine is mutated into serine, so that the mutant amino acid sequence is shown as SEQ ID NO. 54.
8. The lactone hydrolase mutant genetic engineering bacteria, characterized in that the genetic engineering bacteria take escherichia coli or saccharomycetes as host cells to carry out recombinant expression on the lactone hydrolase mutant according to any one of claims 1 to 7.
9. The lactone hydrolase mutants of claim 8, as a class of biocatalysts characterized by: the biocatalyst is microorganism whole cell or enzyme solution containing lactone hydrolase mutant obtained by fermenting the lactone hydrolase gene engineering bacteria of claim 8.
10. The use of the lactone hydrolase mutant as a class of biocatalysts as claimed in claim 9 wherein: the lactone hydrolase mutant is used for splitting D, L-pantoic acid lactone to prepare optically pure D-pantoic acid.
CN202011062911.9A 2020-09-30 2020-09-30 Lactone hydrolase mutant and application thereof Active CN112175919B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202011062911.9A CN112175919B (en) 2020-09-30 2020-09-30 Lactone hydrolase mutant and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202011062911.9A CN112175919B (en) 2020-09-30 2020-09-30 Lactone hydrolase mutant and application thereof

Publications (2)

Publication Number Publication Date
CN112175919A true CN112175919A (en) 2021-01-05
CN112175919B CN112175919B (en) 2024-02-09

Family

ID=73949254

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202011062911.9A Active CN112175919B (en) 2020-09-30 2020-09-30 Lactone hydrolase mutant and application thereof

Country Status (1)

Country Link
CN (1) CN112175919B (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113046337A (en) * 2021-03-18 2021-06-29 赤峰制药股份有限公司 Pantolactone hydrolase mutant strain and application thereof
CN117070494A (en) * 2023-10-13 2023-11-17 天津凯莱英生物科技有限公司 Esterase mutant and application thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1761742A (en) * 2003-03-03 2006-04-19 第一精密化学株式会社 Process for producing lactonase and utilization thereof
CN1793321A (en) * 2005-11-22 2006-06-28 浙江杭州鑫富药业股份有限公司 Microorganism of producing D-pantothenic acid enternal ester hydrolase and process for preparing D-pantothenic acid thereof
CN108004291A (en) * 2017-12-21 2018-05-08 浙江新和成股份有限公司 One kind is used to hydrolyze D, the microbes producing cellulase of L- pantoic acid lactones and its application and screening technique

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1761742A (en) * 2003-03-03 2006-04-19 第一精密化学株式会社 Process for producing lactonase and utilization thereof
CN1793321A (en) * 2005-11-22 2006-06-28 浙江杭州鑫富药业股份有限公司 Microorganism of producing D-pantothenic acid enternal ester hydrolase and process for preparing D-pantothenic acid thereof
CN108004291A (en) * 2017-12-21 2018-05-08 浙江新和成股份有限公司 One kind is used to hydrolyze D, the microbes producing cellulase of L- pantoic acid lactones and its application and screening technique

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MA, L.J., 等: ""XP_018760988.1"", NCBI GENBANK *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113046337A (en) * 2021-03-18 2021-06-29 赤峰制药股份有限公司 Pantolactone hydrolase mutant strain and application thereof
CN113046337B (en) * 2021-03-18 2023-04-07 赤峰制药股份有限公司 Pantolactone hydrolase mutant strain and application thereof
CN117070494A (en) * 2023-10-13 2023-11-17 天津凯莱英生物科技有限公司 Esterase mutant and application thereof
CN117070494B (en) * 2023-10-13 2024-01-19 天津凯莱英生物科技有限公司 Esterase mutant and application thereof

Also Published As

Publication number Publication date
CN112175919B (en) 2024-02-09

Similar Documents

Publication Publication Date Title
US10837036B2 (en) Method for preparing L-aspartic acid with maleic acid by whole-cell biocatalysis
CN111979163B (en) Recombinant Roche bacterium, preparation method and application thereof
CN110938616B (en) Mutant of nitrile hydratase derived from hot spring thermokalite bacillus
CN112877307B (en) Amino acid dehydrogenase mutant and application thereof
CN109182298B (en) Recombinant lipase mutant, engineering bacterium and application
CN112175919A (en) Lactone hydrolase mutant and application thereof
CN114317508B (en) Halohydrin dehalogenase mutant, engineering bacterium and application thereof
CN114480512B (en) Application of oxidoreductase and mutant thereof in biosynthesis of nootkatone
CN115873836A (en) Nerolidol synthetase and application thereof
CN109593749B (en) Halogen alcohol dehalogenase mutant and application thereof in synthesis of chiral epichlorohydrin
CN112175839B (en) D-pantoic acid lactone hydrolase, producing strain, genetic engineering strain and application thereof
CN106754818B (en) Heat-resistant esterase mutant and preparation method and application thereof
CN103981160A (en) Thermomyces lanuginosus lipase mutant, coding genes, and applications of thermomyces lanuginosus lipase mutant
CN105296513B (en) A kind of ocean esterase and its encoding gene E22 and application
CN109609479B (en) Aspergillus usamii epoxide hydrolase mutant with improved enantioselectivity
CN110819609B (en) Mutant lipase with improved thermal stability as well as preparation method and application thereof
CN104560912B (en) Esterase and its encoding gene and application
CN109762801B (en) Halogen alcohol dehalogenase mutant and application thereof in synthesizing chiral drug intermediate
CN108018265B (en) Inositol oxidase mutant and coding gene and application thereof
CN112941003A (en) Method for synthesizing L-alanine by catalyzing maleic acid through double-enzyme coupling whole cells
CN114196659B (en) Amidase mutant, coding gene, engineering bacteria and application thereof
CN113930402B (en) Method for improving laccase catalytic activity, mutant Lcc9-M2, gene and application
CN107236718B (en) Low-temperature esterase from metagenome, coding gene and application thereof
CN114292825B (en) Synthesis method of tropinone
CN114891763B (en) Streptomyces fei-derived cyclooxygenase, gene, vector, engineering bacteria, preparation method and application

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant