CN111699247A - 通过发酵产生(6e)-8-羟基香叶醇的经改造的生物合成途径 - Google Patents

通过发酵产生(6e)-8-羟基香叶醇的经改造的生物合成途径 Download PDF

Info

Publication number
CN111699247A
CN111699247A CN201880088768.5A CN201880088768A CN111699247A CN 111699247 A CN111699247 A CN 111699247A CN 201880088768 A CN201880088768 A CN 201880088768A CN 111699247 A CN111699247 A CN 111699247A
Authority
CN
China
Prior art keywords
leu
ala
ser
val
gly
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.)
Pending
Application number
CN201880088768.5A
Other languages
English (en)
Inventor
C·特雷西韦尔
S·埃德加
S·季莫申科
A·希勒
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.)
Zymergen Inc
Original Assignee
Zymergen Inc
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 Zymergen Inc filed Critical Zymergen Inc
Publication of CN111699247A publication Critical patent/CN111699247A/zh
Pending legal-status Critical Current

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
    • 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
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • 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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • C12N9/0073—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14) with NADH or NADPH as one donor, and incorporation of one atom of oxygen 1.14.13
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
    • 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)
    • 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
    • C12N9/90—Isomerases (5.)
    • 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
    • C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
    • C12P5/007—Preparation of hydrocarbons or halogenated hydrocarbons containing one or more isoprene units, i.e. terpenes
    • 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
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/18—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic polyhydric
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01034—Hydroxymethylglutaryl-CoA reductase (NADPH) (1.1.1.34)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y205/00—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5)
    • C12Y205/01—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5) transferring alkyl or aryl groups, other than methyl groups (2.5.1)
    • C12Y205/01001—Dimethylallyltranstransferase (2.5.1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y205/00—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5)
    • C12Y205/01—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5) transferring alkyl or aryl groups, other than methyl groups (2.5.1)
    • C12Y205/0101—(2E,6E)-Farnesyl diphosphate synthase (2.5.1.10), i.e. geranyltranstransferase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/07—Diphosphoric monoester hydrolases (3.1.7)
    • C12Y301/07011—Geranyl diphosphate diphosphatase (3.1.7.11)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y503/00—Intramolecular oxidoreductases (5.3)
    • C12Y503/03—Intramolecular oxidoreductases (5.3) transposing C=C bonds (5.3.3)
    • C12Y503/03002—Isopentenyl-diphosphate DELTA-isomerase (5.3.3.2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
    • C12Y114/13—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with NADH or NADPH as one donor, and incorporation of one atom of oxygen (1.14.13)
    • C12Y114/13152—Geraniol 8-hydroxylase (1.14.13.152)

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Botany (AREA)
  • Virology (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

本公开内容描述了改造微生物细胞以用于(6E)‑8‑羟基香叶醇的发酵生产，并提供了新的经改造的微生物细胞和培养物，以及相关的(6E)‑8‑羟基香叶醇生产方法。

Description

通过发酵产生(6E)-8-羟基香叶醇的经改造的生物合成途径
相关申请的交叉引用
本申请要求2017年12月7日提交的美国临时申请62/596,013的权益，该临时申请以引用的方式全文纳入本文。
关于在联邦资助的研究和开发下进行的发明的权利的声明
本发明在DARPA授予的协议号HR0011-15-9-0014下的政府支持下进行。政府对本发明拥有一定的权利。
以引用的方式纳入序列表
本申请包括序列表，该序列表以ASCII格式电子提交并且以引用的方式全文纳入本文。该ASCII副本于2018年12月5日生成，被命名为2018-12-05_ZMGNP007WO_SeqList_ST25.txt，大小为327,680字节。
技术领域
本公开内容总体上涉及改造微生物以通过发酵产生(6E)-8-羟基香叶醇的领域。
背景技术
已知(6E)-8-羟基香叶醇(8-羟基香叶醇)是存在于自然界中的无环单萜。已知通过化学合成生产萜烯醇的方法(美国专利4,107,219)。(6E)-8-羟基香叶醇来自基于核心代谢物前体乙酰CoA的甲羟戊酸生物合成途径(图1)。甲羟戊酸生物合成途径中的HMG-CoA还原酶受到反馈抑制[1，2]。8-羟基香叶醇是单萜吲哚生物碱[3]和单萜糖苷的前体(JP2013158298A[4]；美国专利9,518,282)。萜烯已被用于制备新的聚酯材料[5]。已制备了热封萜烯聚合物膜(美国专利3,278,646)。氢化萜烯已被用于聚合物共混物(美国专利3,361,849)。已制备了萜烯树脂和萜烯酚树脂，并将其用作汽车工业的涂层保护膜(美国专利5,643,676)。萜烯已被掺入具有高防潮性能的定向聚丙烯薄膜中，用作包装薄膜材料(美国专利5,500,282)。
发明内容
本公开内容提供了经改造的微生物细胞、微生物细胞的培养物以及生产(6E)-8-羟基香叶醇的方法，包括以下实施方案：
实施方案1：一种经改造的微生物细胞，其中所述经改造的微生物细胞表达：(a)非天然的香叶基二磷酸二磷酸酶(香叶醇合酶)；和(b)非天然的香叶醇-8-羟化酶；其中所述经改造的微生物细胞产生(6E)-8-羟基香叶醇。
实施方案2：实施方案1的经改造的微生物细胞，其中所述经改造的微生物细胞包括一种或多种上游(6E)-8-羟基香叶醇途径酶或上游途径活性调节因子的提高的活性，所述提高的活性为相对于对照细胞的提高。
实施方案3：实施方案2的经改造的微生物细胞，其中所述一种或多种上游(6E)-8-羟基香叶醇途径酶选自ATP-柠檬酸合酶、乙酰CoA合酶、硫解酶、羟甲基戊二酰辅酶A合酶(HMG-CoA合酶)、羟甲基戊二酰辅酶A还原酶(HMG-CoA还原酶)、甲羟戊酸激酶、磷酸甲羟戊酸激酶、二磷酸甲羟戊酸脱羧酶、异戊烯基二磷酸δ-异构酶(isopentenyl-diphosphatedelta-isomerase)和香叶基二磷酸合酶。
实施方案4：实施方案3的经改造的微生物细胞，其中所述一种或多种上游(6E)-8-羟基香叶醇途径酶包含异戊烯基二磷酸δ-异构酶。
实施方案5：实施方案1-4中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶的降低的活性，所述降低的活性为相对于对照细胞的降低。
实施方案6：实施方案5的经改造的微生物细胞，其中所述消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶包括双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶(dimethylallyltranstransferase)和/或香叶基焦磷酸合酶。
实施方案7：实施方案1-6中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞还表达反馈去调控的HMG-CoA还原酶。
实施方案8：实施方案1-7中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括相对于对照细胞，提高的乙酰CoA的可用性，这是由于乙酰CoA合成率较高和/或乙酰CoA降解率较低。
实施方案9：一种经改造的微生物细胞，其中所述经改造的微生物细胞包括：(a)表达非原生的天然香叶基二磷酸二磷酸酶(香叶醇合酶)的方式；和(b)表达非天然的香叶醇-8羟化酶的方式；其中所述经改造的微生物细胞产生(6E)-8-羟基香叶醇。
实施方案10：实施方案9的经改造的微生物细胞，其中所述经改造的微生物细胞包括提高一种或多种上游(6E)-8-羟基香叶醇途径酶或上游途径活性的调节因子活性的方式。
实施方案11：实施方案10的经改造的微生物细胞，其中所述一种或多种上游(6E)-8-羟基香叶醇途径酶选自ATP-柠檬酸合酶、乙酰CoA合酶、硫解酶、羟甲基戊二酰辅酶A合酶(HMG-CoA合酶)、羟甲基戊二酰辅酶A还原酶(HMG-CoA还原酶)、甲羟戊酸激酶、磷酸甲羟戊酸激酶、二磷酸甲羟戊酸脱羧酶、异戊烯基二磷酸δ-异构酶和香叶基二磷酸合酶。
实施方案12：实施方案11的经改造的微生物细胞，其中所述一种或多种上游(6E)-8-羟基香叶醇途径酶包括异戊烯基二磷酸δ-异构酶。
实施方案13：实施方案9-12中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括降低消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶的活性的方式，所述降低的活性为相对于对照细胞的降低。
实施方案14：实施方案13的经改造的微生物细胞，其中所述消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶包括双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶和/或香叶基焦磷酸合酶。
实施方案15：实施方案9-14中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞还包括表达反馈去调控的HMG-CoA还原酶的方式。
实施方案16：实施方案9-15中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括相对于对照细胞，由于乙酰CoA合成速率较高和/或乙酰CoA降解速率较低而提高乙酰CoA可用性的方式。
实施方案17：实施方案1-16中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括真菌细胞。
实施方案18：实施方案17的经改造的微生物细胞，其中所述经改造的微生物细胞包括酵母细胞。
实施方案19：实施方案18的经改造的微生物细胞，其中所述酵母细胞为酵母属(Saccharomyces)细胞。
实施方案20：实施方案19的经改造的微生物细胞，其中所述酵母细胞为酿酒酵母(Saccharomyces cerevisiae)种的细胞。
实施方案21：实施方案18的经改造的微生物细胞，其中所述酵母细胞为亚罗酵母属(Yarrowia)细胞。
实施方案22：实施方案21的经改造的微生物细胞，其中所述酵母细胞为解脂亚罗酵母(Yarrowia lipolytica)种的细胞。
实施方案23：实施方案1-22中任一项的经改造的微生物细胞，其中所述非天然的香叶醇合酶包括与来自紫苏(Perilla setoyensis)的香叶醇合酶具有至少70％的氨基酸序列同一性的香叶醇合酶。
实施方案24：实施方案1-22中任一项的经改造的微生物细胞，其中所述非天然的香叶醇合酶包括与来自葡萄(Vitis vinifera)的香叶醇合酶具有至少70％的氨基酸序列同一性的香叶醇合酶。
实施方案25：实施方案1-23中任一项的经改造的微生物细胞，其中所述非天然的香叶醇-8-羟化酶包括与来自赤豆(Phaseolus angularis)的香叶醇-8-羟化酶具有至少70％的氨基酸序列同一性的香叶醇-8-羟化酶。
实施方案26：实施方案4或12-25中任一项的经改造的微生物细胞，其中所述异戊烯基二磷酸δ-异构酶的提高的活性是通过异源表达异戊烯基二磷酸δ-异构酶来实现的。
实施方案27：实施方案26的经改造的微生物细胞，其中所述异源异戊烯基二磷酸δ-异构酶包括与来自酿酒酵母的异戊烯基二磷酸δ-异构酶具有至少70％氨基酸序列同一性的异戊烯基二磷酸δ-异构酶。
实施方案28：实施方案6和14-27中任一项的经改造的微生物细胞，其中所述消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶包括双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶。
实施方案29：实施方案28的经改造的微生物细胞，其中双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶与来自大肠杆菌(Escherichia coli)的双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶具有至少70％的氨基酸同一性，并包含氨基酸置换S80F。
实施方案30：实施方案7或15-27中任一项的经改造的微生物细胞，其中所述HMG-CoA还原酶为酿酒酵母HMG-CoA还原酶的变体。
实施方案31：实施方案1-30中任一项的经改造的微生物细胞，其中，当培养时，所述经改造的微生物细胞产生水平高于100μg/L培养基的(6E)-8-羟基香叶醇。
实施方案32：实施方案1-31中任一项的经改造的微生物细胞的培养物。
实施方案33：实施方案32的培养物，其中所述底物包括碳源和氮源，所述氮源选自尿素、铵盐、氨及其任意组合。
实施方案34：实施方案32-33中任一项的培养物，其中所述经改造的微生物细胞以一定浓度存在，从而使得所述培养物在600nm下的光密度为10-500。
实施方案35：实施方案32-34中任一项的培养物，其中所述培养物包含(6E)-8-羟基香叶醇。
实施方案36：实施方案32-35中任一项的培养物，其中所述培养物包含水平高于100μg/L培养基的(6E)-8-羟基香叶醇。
实施方案37：培养实施方案1-31中任一项的经改造的微生物细胞的方法，所述方法包括在适合产生(6E)-8-羟基香叶醇的条件下培养细胞。
实施方案38：实施方案37的方法，其中所述方法包括分批补料培养，初始葡萄糖水平范围为1-100g/L，然后是受控的糖补料。
实施方案39：实施方案37或实施方案38的方法，其中所述发酵底物包括葡萄糖和选自尿素、铵盐、氨及其任意组合的氮源。
实施方案40：实施方案37-39中任一项的方法，其中在培养过程中所述培养物是pH受控的。
实施方案41：实施方案37-40中任一项的方法，其中在培养过程中给所述培养物通气。
实施方案42：实施方案37-41中任一项的方法，其中所述经改造的微生物细胞以高于100μg/L培养基的水平产生(6E)-8-羟基香叶醇。
实施方案43：实施方案37-42中任一项的方法，其中所述方法还包括从培养物中回收(6E)-8-羟基香叶醇。
实施方案44：利用经改造以产生(6E)-8-羟基香叶醇的微生物细胞来制备(6E)-8-羟基香叶醇的方法，所述方法包括：(a)在微生物细胞中表达非天然的香叶基二磷酸二磷酸酶(香叶醇合酶)；(b)在微生物细胞中表达非天然的香叶醇-8-羟化酶；(c)在使微生物细胞产生(6E)-8-羟基香叶醇的条件下，在适当的培养基中培养微生物细胞，其中(6E)-8-羟基香叶醇被释放到培养基中；和(d)从培养基中分离(6E)-8-羟基香叶醇。
附图说明
图1：通过发酵产生(6E)-8-羟基香叶醇的途径。
图2：第一轮经改造的宿主酿酒酵母发酵之后，在细胞外培养液中测定的(6E)-8-羟基香叶醇效价。(另见实施例1，表1)
图3：第二轮经改造的宿主酿酒酵母发酵之后，在细胞外培养液中测定的(6E)-8-羟基香叶醇效价。(另见实施例1，表2)
图4：第一轮经改造的宿主解脂亚罗酵母发酵之后，在细胞外培养液中测定的(6E)-8-羟基香叶醇效价。(另见实施例2，表4)
图5：在用于宿主评估的酿酒酵母发酵之后，在细胞外培养液中测定的(6E)-8-羟基香叶醇效价。(另见实施例3，表5)
图6：在用于第四(提高)轮酿酒酵母菌株发酵之后，在细胞外培养液中测定的(6E)-8-羟基香叶醇效价。(另见实施例1，表3)
图7：将启动子-基因-终止子整合到酿酒酵母和解脂亚罗酵母中。
图8：酿酒酵母和解脂亚罗酵母中的启动子替代。
图9：酿酒酵母和解脂亚罗酵母中的靶基因缺失。
具体实施方式
通过生物发酵生产二醇(如(6E)-8-羟基香叶醇)可以使单体经济地用于旧的和新发现的材料应用。含二醇的聚合物具有吸引人的特性，可用于新的材料应用。
我们对代谢[1]进行了检索，以鉴定在工业宿主生物体中能够使代谢途径产生(6E)-8-羟基香叶醇的酶(见表1)。为了改造生产(6E)-8-羟基香叶醇，工业微生物需要基因工程工具和方法来操纵DNA序列(见图7-9)。然后，通过使用单基因和多基因修饰的重复高通量(HTP)菌株改造，系统地重构微生物代谢以产生(6E)-8-羟基香叶醇，包括在并非其所有生化反应或代谢调节模式都已被表征的工业宿主中(参见例如共有和共同未决的美国专利公开20170159045中的HTP菌株改造方法；还可参见共有和共同未决的美国申请62/455,428，说明了生产酪胺的微生物的改造)。
如上所述，(6E)-8-羟基香叶醇是由类萜途径代谢产生的单萜。微生物中有两种类萜生物合成途径：甲羟戊酸途径和非甲羟戊酸途径。酿酒酵母和解脂亚罗酵母都使用甲羟戊酸途径生产萜烯[2]。
本公开内容描述了微生物细胞的改造，用于发酵产生(6E)-8-羟基香叶醇，并提供了新的经改造的微生物细胞和培养物，以及相关的(6E)-8-羟基香叶醇生产方法。
定义
除非另外具体说明，权利要求书和说明书中使用的术语如下文所定义。
术语“发酵”在本文中用于指这样的过程，即其中微生物细胞通过一个或多个生物转化步骤将一种或多种底物转化为所需产物(例如(6E)-8-羟基香叶醇)，而不需要任何化学转化步骤。
本文中使用的术语“经改造的(engineered)”与细胞有关，用于表示所述细胞包含至少一种人工引入的靶向遗传改变，该遗传改变使经改造的细胞区别于天然存在的细胞。
术语“天然的”在本文中用于指天然存在于特定细胞中的细胞组分，例如多核苷酸或多肽。天然多核苷酸或多肽是细胞内源性的。
当与多核苷酸或多肽有关时，术语“非天然的”用于指非天然存在于特定细胞中的多核苷酸或多肽。
当与基因表达的环境有关时，术语“非天然的”用于指在其天然表达的基因组和细胞环境之外的任何环境中中表达的基因。以非天然方式表达的基因可能与宿主细胞中对应基因具有相同的核苷酸序列，但可能从载体表达或从基因组中与天然基因的基因座不同的整合点表达。
术语“异源的”在本文中用于描述被引入到宿主细胞中的多核苷酸或多肽。该术语包含分别来自不同于宿主细胞的生物体、物种或菌株的多核苷酸或多肽。在这种情况下，异源的多核苷酸或多肽的序列不同于同一宿主细胞中存在的任何序列。然而，该术语还包含其序列与在宿主细胞中发现的序列相同的多核苷酸或多肽，其中所述多核苷酸或多肽存在于与天然序列不同的环境下(例如，异源多核苷酸可以连接到不同的启动子上，并插入到与天然序列不同的基因组位置)。因此，“异源表达”包括宿主细胞非天然序列的表达，以及在非天然环境下宿主细胞天然序列的表达。
关于多核苷酸或多肽，术语“野生型”指具有来自天然存在的生物体的多核苷酸或多肽中存在的核苷酸序列或氨基酸序列的任何多核苷酸或任何多肽，无论分子来源如何；即，术语“野生型”指序列特征，无论所述分子是从天然来源中纯化；重组表达，然后纯化；还是合成的。术语“野生型”还用于指天然存在的细胞。
“对照细胞”是这样的细胞，其在其他方面与测试的经改造的细胞相同，包括与经改造的细胞为相同属和种，但是缺少在经改造的细胞中测试的特定遗传修饰。
在本文中，酶是通过它们催化的反应来鉴定，除非另外说明，酶指的是能够催化所鉴定的反应的任何多肽。除非另外说明，酶可源自任何生物体，并且可具有天然的或突变的氨基酸序列。本领域中公知，酶可以具有多种功能和/或多个名称，有时取决于它们所源自的生物体。本文使用的酶涵盖了直系同源物，包括可具有一种或多种额外的功能或不同名称的酶。
术语“反馈去调控的(feedback-deregulated)”在本文中与在特定细胞中通常被酶通路的下游产物负调控(即反馈抑制)的酶有关。在这种情况下，“反馈去调控的”酶是这种形式的酶，其与细胞原生的天然酶相比对反馈抑制更加不敏感。反馈去调控的酶可以通过将一种或多种突变引入到天然酶中而产生。或者，反馈去调控的酶可以仅仅为异源天然酶，当其被引入到特定微生物细胞中时，对反馈抑制的敏感度不如原生天然酶。在一些实施方案中，反馈去调控的酶在微生物细胞中不显示出反馈抑制。
术语“(6E)-8-羟基香叶醇”是指(2E，6E)-2，6-二甲基-2，6-辛二烯-1，8-二醇(CAS#26488-97-1)。
在两个或多个氨基酸或核苷酸序列的情况下，术语“序列同一性”指当进行比较并最大对应的比对时，相同或具有特定的相同氨基酸残基或核苷酸百分比的两个或多个序列，如使用序列比较算法或通过目测来测量的。
对于序列比较以确定百分比核苷酸或氨基酸序列同一性，通常一个序列作为“参考序列”，将“测试”序列与之进行比较。当使用序列比较算法时，将测试序列和参考序列输入到计算机中，如果必要，指定子序列坐标，并且指定序列算法程序参数。然后基于指定的程序参数，序列比较算法计算测试序列相对于参考序列的百分比序列同一性。使用设置为默认参数的BLAST，可以进行序列比对以进行比较。
本文使用的术语“效价”指由微生物细胞培养物产生的产物(例如(6E)-8-羟基香叶醇)的质量除以培养物体积。
关于从细胞培养物中回收(6E)-8-羟基香叶醇，本文使用的“回收”指从至少一种细胞培养基的其他组分中分离出(6E)-8-羟基香叶醇。
改造微生物用于(6E)-8-羟基香叶醇生产
(6E)-8-羟基香叶醇生物合成途径
基于核心代谢物前体乙酰CoA，从甲羟戊酸生物合成途径获得(6E)-8-羟基香叶醇。该途径在图1举例说明。甲羟戊酸生物合成途径中的HMG-CoA还原酶受到反馈抑制。许多微生物(如酿酒酵母)缺乏催化这一途径的最后两个步骤的酶，即香叶基二磷酸二磷酸酶(香叶醇合酶)和香叶醇-8-羟化酶。在这样的微生物宿主中产生(6E)-8-羟基香叶醇需要添加至少一种异源香叶醇合酶和至少一种异源香叶醇-8-羟化酶。
对微生物生产(6E)-8-羟基香叶醇的改造
可将在被改造的微生物细胞中具备活性的任何香叶醇合酶和香叶醇-8-羟化酶引入到细胞中，通常通过使用标准的基因工程技术引入和表达编码所述酶的基因。适合的香叶醇合酶和香叶醇-8-羟化酶可来自任何来源，包括植物、古细菌、真菌、革兰氏阳性细菌和革兰氏阴性细菌来源。示例性来源包括但不限于：紫苏、赤豆、葡萄(葡萄)、川西獐牙菜(Swertia mussotii)(龙胆科植物(Felwort))、毛果杨(Populus trichocarpa)(毛果杨(Western balsam poplar))(香脂杨亚种毛果杨(Populus balsamiferasubsp.Trichocarpa))、罂粟(Papaver somniferum)、欧芹(Petroselinum crispum)、稻(Oryza sativa)、Methanosphaerula palustris、詹氏甲烷球菌(Methanocaldococcusjannaschii)、拜氏接合酵母(Zygosaccharomyces bailii)、马尔尼菲青霉菌(Penicilliummarneffei)、柄篮状菌(Talaromyces stipitatus)、马毛藓菌(Trichophyton equinum)、口丙酸杆菌(Propionibacterium sp.oral)、粪肠球菌(Enterococcus faecium)、吸水链霉菌(Streptomyces hygroscopicus)、斯维链霉菌(Streptomyces sviceus)、海洋贫养杆菌(Modestobacter marinus)、恶臭假单胞菌(Pseudomonas putida)、费氏中华根瘤菌(Sinorhizobium fredii)、Cathatanthus roseaus、玉蜀黍(Zea mays)、长春花(Catharanthus roseus)(长春花(Madagascar periwinkle))(长春花(Vinca rosea))、卷曲回回苏(Perilla frutescens var.crispa)、Perilla frutescens var.hirtella、Cinnamomum tenuipile(Alseodaphne mollis)、罗勒(Ocimum basilicum)(甜罗勒(Sweetbasil))、Perilla citriodora、油橄榄(Olea europaea)(油橄榄(Common olive))、甜舌草(Phyla dulcis)(甜舌草(Aztec sweet herb))(Lippia dulcis)、玫瑰(Rosa rugosa)(玫瑰(Rugosa rose))、喜树(Camptotheca acuminata)(喜树(Happy tree))、粗皮柠檬(Citrus jambhiri)(粗皮柠檬(Rough lemon))、库洛胡黄连(Picrorhiza kurrooa)、拟南芥(Arabidopsis thaliana)(Mouse-ear cress)、大豆(Glycine max)(大豆(Soybean))(大豆(Glycine hispida))、甜菜(Beta vulgaris)(甜菜(Sugar beet))、种棱粟米草(Mollugoverticillata)(种棱粟米草(Green carpetweed))、无油樟(Amborella trichopoda)、马铃薯(Solanum tuberosum)(马铃薯(Potato))、野生大豆(Glycine soja)(野生大豆(Wildsoybean))、大花万带兰(Vanda coerulea)、短舌野生稻(Orvza barthii)、观果金丝桃(Hypericum androsaemum)(Tutsan)、番茄(Solanum lycopersicum)(番茄(Tomato))(番茄(Lycopersicon esculentum))和中果咖啡(Coffea canephora)(罗布斯塔咖啡(Robustacoffee))。
可将香叶醇合酶和香叶醇-8-羟化酶基因的一个或多个拷贝引入选择的微生物宿主细胞。如果引入多于一个基因拷贝，则所述拷贝可以是具有相同或不同核苷酸序列的拷贝。在一些实施方案中，从强的组成型启动子表达一种或两种异源基因。在一些实施方案中，从诱导型启动子表达异源的香叶醇合酶和/或异源的香叶醇-8-羟化酶基因。异源基因可以任选地进行密码子优化，以增强在所选微生物宿主细胞中的表达。
在实施例1中，酿酒酵母经改造以表达来自紫苏的香叶醇合酶(UniProt IDC0KWV4)(SEQ ID NO：5)和来自赤豆的香叶醇-8-羟化酶(UniProt ID C6J436)(SEQ ID NO：11)，在第一轮基因工程中产生的(6E)-8-羟基香叶醇效价为37.5μg/L(下文表1)。在第二轮的菌株中，该效价增加到122.9μg/L，该菌株还表达了三个拷贝的异戊烯基二磷酸δ3-δ2-异构酶(UniProt ID P15496)(SEQ ID NO：25)。
在实施例2中，解脂亚罗酵母经改造以表达来自紫苏的香叶醇合酶(UniProt IDC0KWV4)(SEQ ID NO：99)、来自赤豆的香叶醇8-羟化酶(UniProt ID A0A0L9UT99)(SEQ IDNO：115)和来自酿酒酵母的异戊烯基二磷酸δ3-δ2-异构酶(UniProt ID P15496)(SEQ IDNO：126)，产生的(6E)-8-羟基香叶醇的效价为310微克/L。
在实施例3中，酿酒酵母经改造以表达来自紫苏的香叶醇合酶(UniProt IDC0KWV4)(SEQ ID NO：99)、来自赤豆的香叶醇-8-羟化酶(UniProt ID C6J436)和来自酿酒酵母的异戊烯基二磷酸δ3-δ2-异构酶(UniProt ID P15496)(SEQ ID NO：126)，产生的(6E)-8-羟基香叶醇的效价为217微克/L。
提高上游酶的活性
在能够产生(6E)-8-羟基香叶醇的微生物细胞中提高(6E)-8-羟基香叶醇产量的一个方法是提高(6E)-8-羟基香叶醇生物合成途径中的一种或多种上游酶的活性。上游途径酶包括从进料直到最终天然代谢物(即酿酒酵母中的香叶基二磷酸)的转化中涉及的所有酶。在某些实施方案中，上游途径酶具体指产生(6E)-8-羟基香叶醇的途径中关键前体转化为香叶基二磷酸的过程中涉及的酶。这样的基因包括编码以下酶的基因：ATP-柠檬酸合酶、乙酰CoA合酶、硫解酶、羟甲基戊二酰辅酶A合酶(HMG-CoA合酶)、羟甲基戊二酰辅酶A还原酶(HMG-CoA还原酶)、甲羟戊酸激酶、磷酸甲羟戊酸激酶、二磷酸甲羟戊酸脱羧酶、异戊烯基二磷酸δ-异构酶(系统名称：异戊烯基二磷酸δ2-δ3-异构酶)和香叶基二磷酸合酶。适合的上游途径基因可来自任何来源，包括例如，上文中作为异源香叶醇合酶或香叶醇-8-羟化酶基因的来源被讨论的那些。
在一些实施方案中，通过调节天然酶的表达或活性来提高一种或多种上游途径酶的活性。这种方法的实例包括：(1)过表达HMG-CoA还原酶和/或组成型过表达香叶基二磷酸合酶，以提高(6E)-8-羟基香叶醇途径前体香叶基二磷酸的水平，和/或(2)过表达ATP-柠檬酸合酶(P53396)和/或乙酰CoA合酶(ACS，Q8ZKF6)，提高乙酰CoA的可用性。
通过上游途径活性的一种或多种天然调节因子，可以提高天然上游途径酶的表达。例如，为了提高类异戊二烯途径酶的表达，人们可以引入并任选地过表达甾醇摄取控制蛋白UPC2的变体(UniProt ID Q12151，来自酿酒酵母S288c，含有G888D或G888R[这些命名表示氨基酸置换，使用标准单字母代码表示氨基酸，第一个字母指野生型残基，最后一个字母指替换残基；数字表示翻译的蛋白中氨基酸置换的位置])[6，7]。甾醇摄取控制蛋白UPC2调节甾醇合成，C端氨基酸置换提高了该转录因子的活性。例如，除了其他甾醇生物合成途径基因外，ERG8上游的UPC2结合元件还使UPC2能够转录激活ERG8[7]。
或者，或另外，使用例如图5所示的技术，一个或多个启动子可以置换天然启动子。在某些实施方案中，置换启动子比天然启动子更强，和/或为组成型启动子。
在一些实施方案中，一种或多种上游途径酶的活性通过将一种或多种相应的基因引入表达香叶醇合酶和香叶醇-8-羟化酶的微生物宿主细胞来补充。实施例1描述了成功改造微生物宿主细胞以表达异源香叶醇合酶和异源香叶醇-8-羟化酶，以及编码异戊烯基二磷酸δ-异构酶的引入基因。
所引入的上游途径基因可能来自宿主细胞以外的生物体，或仅为天然基因的额外拷贝。在一些实施方案中，将一种或多种这样的基因引入到能够产生(6E)-8-羟基香叶醇的微生物宿主细胞中，并从强的组成型启动子表达和/或可任选地对所述基因进行密码子优化，以增强在所选微生物宿主细胞中的表达。
在各实施方案中，改造生产(6E)-8-羟基香叶醇的微生物细胞以提高一种或多种上游途径酶的活性，这将(6E)-8-羟基香叶醇效价提高至少10％、20％、30％、40％、50％、60％、70％、80％或90％，或至少2倍、2.5倍、3倍、3.5倍、4倍、4.5倍、5倍、5.5倍、6倍、6.5倍、7倍、7.5倍、8倍、8.5倍、9倍、9.5倍、10倍、11倍、12倍、13倍、14倍、15倍、16倍、17倍、18倍、19倍、20倍、21倍、22倍、23倍、24倍、25倍、30倍、35倍、40倍、45倍、50倍、55倍、60倍、65倍、70倍、75倍、80倍、85倍、90倍、95倍或100倍。在各实施方案中，(6E)-8-羟基香叶醇效价的提高在10％至100倍、2倍至50倍、5倍至40倍、10倍至30倍的范围内，或以上列出的任何数值界定的任何范围内。(本文的范围包括它们的端点。)这些提高相对于在上游途径酶活性没有任何提高的产(6E)-8-羟基香叶醇微生物细胞中测定的(6E)-8-羟基香叶醇效价而确定。这种参考细胞可具有一种或多种旨在增加(6E)-8-羟基香叶醇产生的其他遗传改变，例如，所述细胞可表达反馈去调控的酶。
在各实施方案中，通过提高一种或多种上游途径基因的活性而实现的(6E)-8-羟基香叶醇效价为至少100、200、300、400、500、600、700、800或900μg/L或至少1、10、50、75、100、200、300、400、500、600、700、800或900mg/L或至少1、1.5、2、2.5、3、3.5、4、4.5、5或10gm/L。在各实施方案中，所述效价在100μg/L至10gm/L、200μg/L至5gm/L、500μg/L至4gm/L、1mg/L至3gm/L、500mg/L至2gm/L的范围内，或以上列出的任何数值界定的任何范围内。
反馈去调控的酶的引入
由于(6E)-8-羟基香叶醇生物合成受到反馈抑制，在经改造以产生(6E)-8-羟基香叶醇的微生物细胞中提高(6E)-8-羟基香叶醇产量的另一种方法是引入通常受到反馈调节的一种或多种酶的反馈去调控形式。HMG-CoA还原酶就是这样一种酶。反馈去调控形式可以是异源、天然酶，其与特定微生物宿主细胞中的天然酶相比对反馈抑制更不敏感。或者，反馈去调控形式可以是天然或异源酶的变体，其具有一个或多个突变或被截短，使其与对应的天然酶相比对反馈抑制更不敏感。后者的实例包括HMG-CoA还原酶变体(来自酿酒酵母)，其具有N端截短(SEQ ID NO：27)。这种反馈去调控的HMG-CoA还原酶在宿主细胞中的表达已被证明能提高酿酒酵母[3]和其他生物体中的甲羟戊酸途径通量(参见例如，PCT公开WO2001031027A1，描述植物基因工程)。
在各实施方案中，改造生产(6E)-8-羟基香叶醇的微生物细胞以表达反馈去调控的酶，这将(6E)-8-羟基香叶醇的效价提高至少10％、20％、30％、40％、50％、60％、70％、80％或90％，或至少2倍、2.5倍、3倍、3.5倍、4倍、4.5倍、5倍、5.5倍、6倍、6.5倍、7倍、7.5倍、8倍、8.5倍、9倍、9.5倍、10倍、11倍、12倍、13倍、14倍、15倍、16倍、17倍、18倍、19倍、20倍、21倍、22倍、23倍、24倍、25倍、30倍、35倍、40倍、45倍、50倍、55倍、60倍、65倍、70倍、75倍、80倍、85倍、90倍、95倍或100倍。在各实施方案中，(6E)-8-羟基香叶醇效价的提高在10％至100倍、2倍至50倍、5倍至40倍、10倍至30倍的范围内，或以上列出的任何数值界定的任何范围内。这些提高相对于在不表达反馈去调控的酶的产(6E)-8-羟基香叶醇微生物细胞中测定的(6E)-8-羟基香叶醇效价而确定。这种参考细胞可以(但是不需要)具有旨在增加(6E)-8-羟基香叶醇产量的其他遗传改变，即所述细胞可具有上游途径酶的提高的活性，这是由反馈不敏感以外的一些方式引起的。
在各实施方案中，通过使用反馈去调控的酶以增加经过(6E)-8-羟基香叶醇生物合成途径的通量，实现的(6E)-8-羟基香叶醇效价为至少100、200、300、400、500、600、700、800或900μg/L，或至少1、10、50、75、100、200、300、400、500、600、700、800或900mg/L或至少1、1.5、2、2.5、3、3.5、4、4.5、5或10g/L。在各实施方案中，效价在100μg/L至10g/L、200μg/L至5g/L、500μg/L至4g/L、1mg/L至3g/L、500mg/L至2g/L的范围内，或以上列出的任何数值界定的任何范围内。
补充一种或多种天然酶的活性和/或引入一种或多种反馈去调控的酶的方法可以在表达香叶醇合酶的微生物细胞中结合以实现甚至更高的(6E)-8-羟基香叶醇产生水平。
减少前体消耗
在能够产生(6E)-8-羟基香叶醇的微生物细胞中增加(6E)-8-羟基香叶醇产量的另一种方法是降低一种或多种酶的活性，所述酶将(6E)-8-羟基香叶醇生物合成的一种或多种前体分流到一个或多个旁路(side pathway)(即产生(6E)-8-羟基香叶醇以外的其他产物的途径)。在一些实施方案中，通过调节天然酶的表达或活性来降低一种或多种旁路酶的活性。示例性旁路酶包括双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶、香叶基焦磷酸合酶和任何消耗乙酰CoA的旁路酶。例如，用活性较低或失活的启动子代替对应基因的天然启动子，或者缺失对应的基因，可以使这些酶的活性降低。参见图5和图6，其分别为酿酒酵母中启动子替换和靶基因缺失的方案的实例。
天然双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶具有双重功能，可形成香叶基二磷酸，随后第二反应可将香叶基二磷酸酯转化为(2E，6E)-法呢基二磷酸。由于天然酶具有这两种活性，中间体为(6E)-8-羟基香叶醇途径代谢物，有利于降低天然双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶的表达。然而，含有S80F氨基酸置换的酶的表达产生了可测量数量的类单萜[4](另见美国专利8,715,962)。例如，为了防止向甾醇的额外通量，酿酒酵母中由ERG 9编码的双功能的法呢基-二磷酸法呢基转移酶(EC2.5.1.21)，和/或酿酒酵母中由ERG 20编码的(2E，6E)-法呢基二磷酸合酶(EC2.5.1.10)的表达或活性降低，以最大化香叶基二磷酸池，用于(6E)-8-羟基香叶醇生物合成。
在酿酒酵母中的示例性实施方案中：(1)双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶(ERG20，YJL167W)的启动子可以替换为酿酒酵母pRnr1启动子，以降低这种消耗(6E)-8-羟基香叶醇途径代谢物香叶基二磷酸的天然酶的表达；(2)香叶基香叶基焦磷酸合酶(Bts1，YPL069C)的启动子可以替换为酿酒酵母pPsp2，以降低这种也消耗香叶基二磷酸的天然酶的表达；和/或可以缺失基因Pdc5(YLR134W)、Pdc6(YGR087C)和Pdc1(YLR044C)中的一个或多个，以减少乙酰CoA的消耗。
在各实施方案中，改造生产(6E)-8-羟基香叶醇的微生物细胞以减少一个或多个旁路对前体的消耗，这将(6E)-8-羟基香叶醇效价提高至少10％、20％、30％、40％、50％、60％、70％、80％或90％，或至少2倍、2.5倍、3倍、3.5倍、4倍、4.5倍、5倍、5.5倍、6倍、6.5倍、7倍、7.5倍、8倍、8.5倍、9倍、9.5倍、10倍、11倍、12倍、13倍、14倍、15倍、16倍、17倍、18倍、19倍、20倍、21倍、22倍、23倍、24倍、25倍、30倍、35倍、40倍、45倍、50倍、55倍、60倍、65倍、70倍、75倍、80倍、85倍、90倍、95倍或100倍。在各实施方案中，(6E)-8-羟基香叶醇效价的提高在10％至100倍、2倍至50倍、5倍至40倍、10倍至30倍的范围内，或以上列出的任何数值界定的任何范围内。这些提高相对于在不包括减少前体消耗的遗传改变的产(6E)-8-羟基香叶醇微生物细胞中测定的(6E)-8-羟基香叶醇效价而确定。这种参考细胞可以(但是不需要)具有旨在增加(6E)-8-羟基香叶醇产量的其他遗传改变，即所述细胞可具有上游途径酶的提高的活性。
在各实施方案中，通过减少一个或多个旁路对前体的消耗，实现的(6E)-8-羟基香叶醇效价为至少100、200、300、400、500、600、700、800或900μg/L，或至少1、10、50、75、100、200、300、400、500、600、700、800或900mg/L或至少1、1.5、2、2.5、3、3.5、4、4.5、5或10g/L。在各实施方案中，效价在100μg/L至10g/L、200μg/L至5gm/L、500μg/L至4g/L、1mg/L至3g/L、500mg/L至2g/L的范围内，或以上列出的任何数值界定的任何范围内。
增加一种或多种天然酶的活性和/或引入一种或多种反馈去调控的酶和/或减少一个或多个旁路对前体消的耗的方法可以结合以实现甚至更高的(6E)-8-羟基香叶醇产生水平。
微生物宿主细胞
可以改造任何可用于表达引入基因的微生物，用于如上文所述发酵生产(6E)-8-羟基香叶醇。在某些实施方案中，微生物是天然无法发酵生产(6E)-8-羟基香叶醇的微生物。在一些实施方案中，微生物是易于培养的微生物，例如，已知适用于作为目的化合物发酵生产中的宿主细胞的微生物。可以如上文所述，改造细菌细胞，包括革兰氏阳性细菌或革兰氏阴性细菌。除谷氨酸棒状杆菌(C.glutamicum)以外，实例还包括枯草芽孢杆菌(Bacillus subtilus)、地衣芽孢杆菌(B.licheniformis)、迟缓芽孢杆菌(B.lentus)、短芽孢杆菌(B.brevis)、嗜热脂肪芽孢杆菌(B.stearothermophilus)、嗜碱芽孢杆菌(B.alkalophilus)、解淀粉芽孢杆菌(B.amyloliquefaciens)、克劳氏芽孢杆菌(B.clausii)、耐盐芽孢杆菌(B.halodurans)、巨大芽孢杆菌(B.megaterium)、凝结芽孢杆菌(B.coagulans)、环状芽孢杆菌(B.circulans)、灿烂芽孢杆菌(B.lautus)、苏云金芽孢杆菌(B.thuringiensis)、白色链霉菌(S.albus)、变铅青链霉菌(S.lividans)、天蓝色链霉菌(S.coelicolor)、灰色链霉菌(S.griseus)、假单胞菌属种(Pseudomonas sp.)、产碱假单胞菌(P.alcaligenes)、柑橘假单胞菌(P.citrea)、乳杆菌属种(Lactobacilis spp.)(例如乳酸乳杆菌(L.lactis)、植物乳杆菌(L.plantarum))、灰色乳杆菌(L.grayi)、大肠杆菌、粪肠球菌、鹑鸡肠球菌(E.gallinarum)、酪黄肠球菌(E.casseliflavus)、和/或粪肠球菌(E.faecalis)细胞。
有多种类型的厌氧细胞可用作本文所述方法中的微生物宿主细胞。在一些实施方案中，微生物细胞是专性厌氧细胞。专性厌氧菌在存在氧的条件下通常无法良好生长(如果不是完全不生长的话)。应理解，可存在少量氧，即专性厌氧菌对低水平的氧具有一定的耐受水平。如上文所述改造的专性厌氧菌可生长在基本无氧的条件下，其中存在的氧的量对厌氧菌的生长、维持和/或发酵无害。
或者，用于本文所述方法的微生物宿主细胞可以是兼性厌氧细胞。如果存在氧，兼性厌氧菌可通过有氧呼吸(例如利用TCA循环)产生细胞ATP。但是，兼性厌氧菌还可在无氧情况下生长。如上文所述改造的兼性厌氧菌可以在基本无氧的条件下生长，其中存在的氧的量对厌氧菌的生长、维持和/或发酵无害，或者可以在存在较大量的氧的情况下生长。
在一些实施方案中，用于本文所述方法的微生物宿主细胞是丝状真菌细胞。(参见，例如Berka&Barnett，Biotechnology Advances，(1989)，7(2)：127-154)。实例包括长枝木霉(Trichoderma longibrachiatum)、绿色木霉(T.viride)、康氏木霉(T.koningii)、哈茨木霉(T.harzianum)、青霉菌属种(Penicillium sp.)、特异腐质霉(Humicolainsolens)、H.lanuginose、灰腐质霉(H.grisea)、金孢子菌属种(Chrysosporium sp.)、C.lucknowense、粘帚霉属种(Gliocladium sp.)、曲霉属种(Aspergillus sp.)(例如，米曲霉(A.oryzae)、黑曲霉(A.niger)、酱油曲霉(A.sojae)、日本曲霉(A.japonicus)、构巢曲霉(A.nidulans)或泡盛曲霉(A.awamori))、镰刀菌属种(Fusarium sp.)(例如，粉红镰刀菌(F.roseum)、禾谷镰刀菌(F.graminum)、谷类镰刀菌(F.cerealis)、F.oxysporuim或F.venenatum)、链孢霉属种(Neurospora sp.)(例如，粗糙链孢霉(N.crassa))或肉座菌属种(Hypocrea sp.)、毛霉属种(Mucor sp.)(例如，白色毛霉(M.miehei))、根霉属种(Rhizopus sp.)、和裸胞壳属种(Emericella sp.)细胞。在具体实施方案中，如上文所述被改造的真菌细胞是构巢曲霉、泡盛曲霉、米曲霉、棘孢曲霉(A.aculeatus)、黑曲霉、日本曲霉、里氏木霉(T.reesei)、绿色木霉、F.oxysporum或腐皮镰刀菌(F.solani)。用于这样的宿主的示例性质粒或质粒组件包括美国专利公开2011/0045563中描述的那些。
酵母还可用作本文所述方法中的微生物宿主细胞。实例包括：酵母属种(Saccharomyces sp.)、裂殖酵母属种(Schizosaccharomyces sp.)，毕赤酵母属种(Pichiasp.)、多形汉逊酵母(Hansenula polymorpha)、叶柄毕赤酵母(Pichia stipites)、马克斯克鲁维酵母(Kluyveromyces marxianus)、克鲁维酵母属种(Kluyveromyces spp.)、解脂亚罗酵母和假丝酵母属种(Candida sp.)。在一些实施方案中，酵母属种是酿酒酵母(参见，例如Romanos et al.，Yeast，(1992)，8(6)：423-488)。用于这类宿主的示例性质粒或质粒组件包括美国专利7,659,097和美国专利公开2011/0045563中描述的那些。
在一些实施方案中，宿主细胞可以是藻类细胞，其来自绿藻、红藻、灰胞藻(glaucophyte)、chlorarachniophyte、眼虫(euglenid)、假菌界(chromista)或鞭毛藻(dinoflagellate)。(参见例如，Saunders&Warmbrodt，“Gene Expression in Algae andFungi，Including Yeast，”(1993)，National Agricultural Library，Beltsville，Md.)。用于藻类细胞的示例性质粒或质粒组件包括美国专利公开2011/0045563中描述的那些。
在其他实施方案中，宿主细胞是蓝细菌，例如根据形态学归类到以下组中任一组的蓝细菌：绿球藻目(Chlorococcales)、宽球藻目(Pleurocapsales)、颤藻目(Oscillatoriales)、念珠藻目(Nostocales)、Synechosystic或真枝藻目(Stigonematales)(参见，例如Lindberg et al.，Metab.Eng.，(2010)12(1)：70-79)。用于蓝细菌细胞的示例性质粒或质粒组件包括美国专利公开2010/0297749和2009/0282545以及国际专利公开WO 2011/034863中描述的那些。
基因工程方法
使用本领域技术范围内的常规的分子生物学(包括重组技术)、微生物学、细胞生物学和生物化学技术，可以改造微生物细胞，用于发酵生产(6E)-8-羟基香叶醇。这类技术在文献中充分解释，参见，例如“Molecular Cloning：A Laboratory Manual，”fourthedition(Sambrook et al.，2012)；“Oligonucleotide Synthesis”(M.J.Gait，ed.，1984)；“Culture of Animal Cells：A Manual of Basic Technique and SpecializedApplications”(R.I.Freshney，ed.，6th Edition，2010)；“Methods in Enzymology”(Academic Press，Inc.)；“Current Protocols in Molecular Biology”(F.M.Ausubel etal.，eds.，1987，and periodic updates)；“PCR：The Polymerase Chain Reaction，”(Mullis et al.，eds.，1994)；Singleton et al.，Dictionary of Microbiology andMolecular Biology 2nd ed.，J.Wiley&Sons(New York，N.Y.1994)。
载体是用于将遗传物质引入到细胞中的多核苷酸载体。适用于本文所述方法的载体可以是线性的或环状的。载体可整合到宿主细胞的靶基因组中或在宿主细胞中独立复制。对于许多应用而言，优选产生稳定转化体的整合载体。载体可包含，例如，复制起点、多克隆位点(MCS)和/或选择性标记。表达载体通常包括含有调控元件的表达盒，所述调控元件促使多核苷酸序列(通常为编码序列)在特定宿主细胞中的表达。载体包括但不限于，整合载体、原核质粒、附加体(episome)、病毒载体、粘粒和人工染色体。
可用于表达盒的示例性调控元件包括启动子、增强子、内部核糖体进入位点(IRES)和其他表达调控元件(例如转录终止信号，如多腺苷酸化信号和poly-U序列)。这类调控元件记载于，例如Goeddel，Gene Expression Technology：Methods In Enzymology185，Academic Press，San Diego，Calif.(1990)。
在一些实施方案中，载体可用于引入能够进行基因组编辑的系统，例如CRISPR系统。参见2014年3月6日公布的美国专利公开2014/0068797；还可参见Jinek M.，et al.，“Aprogrammable dual-RNA-guided DNA endonuclease in adaptive bacterialimmunity，”Science 337：816-21，2012)。在II型CRISPR-Cas9系统中，Cas9是定点核酸内切酶，即使用两个不同的核酸内切酶结构域(HNH和RuvC/类RNase H结构域)可被定向以在特定靶序列切割多核苷酸的酶。可改造Cas9以在任何所需位点切割DNA，因为Cas 9是由RNA定向到其切割位点。因此，Cas9还被描述为“RNA引导的核酸酶”。更具体而言，Cas9与一个或多个RNA分子连接，所述RNA分子将Cas9导向特定的靶多核苷酸，这是基于所述RNA分子的至少一部分与靶多核苷酸的特定序列杂交。Ran，F.A.等人(“In vivo genome editing usingStaphylococcus aureus Cas9，”Nature 520(7546)：186-91，2015，Apr 9]，包括全部扩展数据)提供了crRNA/tracrRNA序列和八种II型CRISPR-Cas9系统的二级结构。类Cas9合成蛋白也是本领域已知的(参见2014年10月23日公开的美国公开专利申请2014-0315985)。
实施例1描述了用于将多核苷酸和其他遗传改变引入到酿酒酵母细胞的基因组中的示例性整合方法。
通过多种标准方法的任一种可将载体或其他多核苷酸引入到微生物细胞中，所述方法例如转化、接合、电穿孔、核微注射、转导、转染(例如，脂质转染介导的或DEAE-糊精介导的转染或使用重组噬菌体病毒的转染)、使用磷酸钙DNA沉淀物孵育、使用DNA包被的微粒高速轰击、和原生质体融合。通过本领域已知的任何方法可选择转化体。适用于选择转化体的方法记载于美国专利公开2009/0203102、2010/0048964和2010/0003716，以及国际公开WO 2009/076676、WO 2010/003007和WO 2009/132220。
经改造的微生物细胞
上述方法可用于产生经改造的微生物细胞，在某些实施方案中，所述微生物细胞过量产生(6E)-8-羟基香叶醇。与天然微生物细胞(例如本文所述的任何微生物宿主细胞)相比，经改造的微生物细胞可具有至少1、2、3、4、5、6、7、8、9、10或更多个遗传改变，例如30-40个改变。下文实施例中描述的经改造的微生物细胞具有1、2或3个遗传改变，但是本领域技术人员按照本文的指导能够设计具有其他改变的微生物细胞。在一些实施方案中，与天然微生物细胞相比，经改造的微生物细胞具有不多于15、14、13、12、11、10、9、8、7、6、5或4个遗传改变。在各实施方案中，经改造以生产(6E)-8-羟基香叶醇的微生物细胞可具有落入以下任何示例性范围的多个遗传改变：1-10、1-9、1-8、2-7、2-6、2-5、2-4、2-3、3-7、3-6、3-5、3-4等。
在一些实施方案中，经改造的微生物细胞表达至少一种异源的香叶基二磷酸二磷酸酶(香叶醇合酶)，如在天然无法产生(6E)-8-羟基香叶醇的微生物宿主细胞的情况下。在各实施方案中，微生物细胞可包含并表达，例如：(1)单一异源香叶醇合酶基因，(2)两种或多种异源香叶醇合酶基因，其可相同或不同(换言之，可引入相同的异源香叶醇合酶基因的多个拷贝，或者可引入多种不同的异源香叶醇合酶基因)，(3)不是细胞原生的单一异源香叶醇合酶基因以及天然香叶醇合酶基因的一个或多个额外拷贝，或(4)两种或多种非天然的香叶醇合酶基因，其可相同或不同，以及天然香叶醇合酶基因的一个或多个额外拷贝。
在一些实施方案中，除了至少一种异源的香叶醇合酶以外，经改造的微生物细胞还表达至少一种异源的香叶醇-8-羟化酶，如在不具有香叶醇-8-羟化酶的微生物宿主细胞的情况下。在各实施方案中，微生物细胞可包含并表达，例如：(1)单一异源香叶醇-8-羟化酶基因，(2)两种或多种异源香叶醇-8-羟化酶基因，其可相同或不同(换言之，可引入相同的异源香叶醇-8-羟化酶基因的多个拷贝，或者可引入多种不同的异源香叶醇-8-羟化酶基因)，(3)不是细胞原生的单一异源香叶醇-8-羟化酶以及天然香叶醇-8-羟化酶基因的一个或多个额外拷贝，或(4)两种或多种非天然香叶醇-8-羟化酶基因，其可相同或不同，以及天然香叶醇-8-羟化酶基因的一个或多个额外拷贝。
这种经改造的宿主细胞可包含至少一种其他遗传改变，其增加经由所述途径产生香叶基-PP((6E)-8-羟基香叶醇的直接前体)的通量。途径中的这些“上游”酶包括：ATP-柠檬酸合酶、乙酰CoA合酶、硫解酶、羟甲基戊二酰辅酶A合酶(HMG-CoA合酶)、羟甲基戊二酰辅酶A还原酶(HMG-CoA还原酶)、甲羟戊酸激酶、磷酸甲羟戊酸激酶、二磷酸甲羟戊酸脱羧酶、异戊烯基二磷酸δ-异构酶和香叶基二磷酸合酶，包括具有这些酶活性的任何同工型(isoform)、旁系同源物或直系同源物(本领域技术人员易于理解，它们可称为不同的名称)。所述至少一种其他改变可通过任何可获得的方式提高上游途径酶的活性，例如通过：(1)调节天然酶的表达或活性，(2)表达天然酶的基因的一个或多个额外拷贝，或(3)表达一种或多种非天然酶的基因的一个或多个拷贝。
在一些实施方案中，可通过表达一种或多种编码反馈去调控的酶的基因来实现经由所述途径的通量的增加，如上文所讨论。例如，经改造的宿主细胞可包含并表达一种或多种反馈去调控的HMG-CoA还原酶基因。
经改造的微生物细胞可包含引入的基因，其具有天然的核苷酸序列或不同于天然的核苷酸序列。例如，可对天然的核苷酸序列进行密码子优化，用于在特定宿主细胞中表达。任何这些引入的基因编码的氨基酸序列可以是天然的或可以不同于天然的序列。在各实施方案中，氨基酸序列与天然的氨基酸序列具有至少0％、70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。
在一些实施方案中，通过降低一种或多种旁路酶(如双功能的(2E，6E)-法尼基二磷酸合酶/二甲基烯丙基反式转移酶、香叶基香叶基焦磷酸合酶和任何消耗乙酰CoA的侧途径酶)的表达或活性，可以实现前体至(6E)-8-羟基香叶醇的提高的可用性。例如，经改造的宿主细胞可以包括一个或多个启动子替换，以下调任何这些酶的表达和/或可以缺失它们的基因，以完全消除它们的表达。
本文所述方法在真菌细胞即酿酒酵母(真核细胞)和细菌细胞即谷氨酸棒状杆菌(原核细胞)中进行。(参见实施例1。)
示例性经改造的真菌细胞
在某些实施方案中，经改造的酵母(例如酿酒酵母)细胞表达异源的香叶醇合酶，其与来自紫苏的香叶醇合酶(如SEQ ID NO：5)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，紫苏香叶醇合酶可包含SEQ ID NO：5。
经改造的酵母(例如酿酒酵母)细胞还表达异源的香叶醇-8-羟化酶，在某些实施方案中，其与来自赤豆的香叶醇-8-羟化酶(如SEQ ID NO：11)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，赤豆香叶醇-8-羟化酶可包含SEQ ID NO：11。
这些可以是经改造的酵母细胞的唯一遗传改变，或者所述酵母细胞可包含一个或多个额外的遗传改变，如上文更一般性的讨论。
具有一个或多个额外的遗传改变的示例性酵母(例如酿酒酵母)细胞相对于对照细胞可具有上游途径酶(例如异戊烯基二磷酸δ-异构酶)的提高的活性，例如通过以下方法而产生：将天然的酿酒酵母异戊烯基二磷酸δ-异构酶(SEQ ID NO：25)基因的额外拷贝引入细胞，或引入编码异戊烯基二磷酸δ-异构酶的基因，该基因与天然的酿酒酵母异戊烯基二磷酸δ-异构酶具有至少70％、75％、80％、85％、90％或95％氨基酸序列同一性。
在具体实施方案中，经改造的酵母(例如酿酒酵母)细胞还表达酿酒酵母HMG-CoA还原酶的变体，其通常与天然的酿酒酵母HMG-CoA还原酶具有至少70％、75％、80％、85％、90％或95％氨基酸序列同一性，或酿酒酵母HMG-CoA还原酶的截短的变体，其中氨基酸残基1-529缺失。在具体实施方案中，酿酒酵母HMG-CoA还原酶变体可包含SEQ ID NO：27。
培养经改造的微生物细胞
可以培养本文所述的任何微生物细胞，例如用于维持、生长和/或产生(6E)-8-羟基香叶醇。
在一些实施方案中，培养物生长到600nm下的光密度为10-500，例如光密度为50-150。
在各实施方案中，培养物包括产生的(6E)-8-羟基香叶醇，其效价为至少10、25、50、75、100、200、300、400、500、600、700、800或900μg/L，或至少1、10、50、75、100、200、300、400、500、600、700、800或900mg/L或至少1、1.5、2、2.5、3、3.5、4、4.5、5或10gm/L。在各实施方案中，效价范围为10μg/L至10gm/L、25μg/L至10gm/L、100μg/L至10gm/L、200μg/L至5gm/L、500μg/L至4gm/L、1mg/L至3gm/L、500mg/L至2gm/L，或以上列出的任何数值界定的任何范围内。
培养基
微生物细胞可以在任何适合的培养基中培养，所述培养基包括但不限于基本培养基，即包含可用于细胞生长的最低营养成分的培养基。基本培养基通常包含：(1)用于微生物生长的碳源；(2)盐，其可取决于具体的微生物细胞和生长条件；和(3)水。适合的培养基还包含以下成分的任意组合：用于生长和产物形成的氮源，用于生长的硫源，用于生长的磷酸盐源，用于生长的金属盐，用于生长的维生素，以及用于生长的其他辅因子。
任何适合的碳源均可用于培养宿主细胞。术语“碳源”指一种或多种能够被微生物细胞代谢的含碳化合物。在各实施方案中，碳源是碳水化合物(例如单糖、二糖、寡糖或多糖)或转化糖(例如酶处理的蔗糖浆)。示例性单糖包括葡萄糖(右旋糖)、果糖(左旋糖)和半乳糖；示例性寡糖包括葡聚糖(dextran或glucan)；示例性多糖包括淀粉和纤维素。适合的糖包括C6糖(例如果糖、甘露糖、半乳糖或葡萄糖)和C5糖(例如木糖或阿拉伯糖)。其他更便宜的碳源包括甘蔗汁、甜菜汁、高粱汁等，任何这些都可以(但不需要)完全或部分去离子化。
培养基中的盐通常提供必需元素，例如镁、氮、磷和硫，以使细胞合成蛋白质和核酸。
基本培养基可补充一种或多种选择性试剂，例如抗生素。
为了产生(6E)-8-羟基香叶醇，培养基可包含和/或在培养中补充葡萄糖和/或氮源，例如尿素、铵盐、氨或其任意组合。
培养条件
适于微生物的维持和生长的材料和方法是本领域公知的。参见，例如美国公布2009/0203102、2010/0003716和2010/0048964，以及国际公布WO 2004/033646、WO 2009/076676、WO 2009/132220和WO 2010/003007，Manual of Methods for GeneralBacteriology，Gerhardt等人编著，American Society for Microbiology，Washington，D.C.(1994)或Brock in Biotechnology：A Textbook 0f Industrial Microbiology，第二版(1989)Sinauer Associates，Inc.，Sunderland，Mass。
通常，细胞生长并维持在适当的温度、气体混合物和pH下(例如约20℃至约37℃，约6％至约84％CO2，pH为约5至约9)。在一些方面中，细胞生长于35℃下。在某些实施方案中，例如当嗜热细菌用作宿主细胞时，可以使用更高的温度(例如50℃-75℃)。在一些方面中，用于发酵的pH范围为约pH 5.0至约pH 9.0(例如约pH 6.0至约pH 8.0或约6.5至约7.0)。根据具体细胞的需求，细胞可生长在有氧、缺氧或厌氧条件下。
可使用的标准培养条件和发酵模式，例如分批发酵、分批补料发酵或连续发酵记载于美国公布2009/0203102、2010/0003716和2010/0048964，以及国际公布WO 2009/076676、WO 2009/132220、和WO 2010/003007中。分批发酵和分批补料发酵是本领域中常见和公知的，实例可参见Brock，Biotechnology：A Textbook of Industrial Microbiology，第二版(1989)Sinauer Associates，Inc.。
在一些实施方案中，将细胞培养在有限糖(例如葡萄糖)的条件下。在各实施方案中，添加的糖的量少于或约为细胞可消耗的糖的量的105％(例如约100％、90％、80％、70％、60％、50％、40％、30％、20％或10％)。在具体实施方案中，添加到培养基的糖的量与特定时间段内被细胞消耗的糖的量大约相同。在一些实施方案中，通过限制添加的糖的量来控制细胞生长速率，从而使细胞以培养基中糖的量能够支持的速率生长。在一些实施方案中，糖在培养细胞期间不积累。在各实施方案中，在有限糖条件下培养细胞，培养时间长于或约为1、2、3、5、10、15、20、25、30、35、40、50、60或70小时，或甚至最长达约5-10天。在各实施方案中，在有限糖的条件下培养细胞，培养时间长于或约为培养细胞的总时长的5、10、15、20、25、30、35、40、50、60、70、80、90、95或100％。尽管不意欲受限于任何具体理论，认为有限糖的条件可以更有利地调节细胞。
在一些方面中，细胞生长于分批培养物中。细胞还可以生长于分批补料培养物或连续培养物中。另外，细胞可生长于基本培养基中，包括但不限于上文所述的任何基本培养基。基本培养基可进一步补充1.0％(w/v)或更少的葡萄糖(或任何其他六碳糖)。特别地，基本培养基可补充1％(w/v)、0.9％(w/v)、0.8％(w/v)、0.7％(w/v)、0.6％(w/v)、0.5％(w/v)、0.4％(w/v)、0.3％(w/v)、0.2％(w/v)或0.1％(w/v)葡萄糖。在一些培养中，使用显著更高水平的糖(例如葡萄糖)，例如至少10％(w/v)、20％(w/v)、30％(w/v)、40％(w/v)、50％(w/v)、60％(w/v)、70％(w/v)，或最高达所述糖在培养基中的溶解度极限。在一些实施方案中，糖水平落入以上数值中任意两个的范围内，例如0.1-10％(w/v)、1.0-20％(w/v)、10-70％(w/v)、20-60％(w/v)或30-50％(w/v)。此外，不同的糖水平可用于不同的培养阶段。对于(例如酿酒酵母或谷氨酸棒状杆菌的)分批补料培养，分批阶段的糖水平可以为约100-200g/L(10-20％(w/v))，之后最高达约500-700g/L(在补料中50-70％)。
另外，基本培养基可补充0.1％(w/v)或更少的酵母提取物。特别地，基本培养基可补充0.1％(w/v)、0.09％(w/v)、0.08％(w/v)、0.07％(w/v)、0.06％(w/v)、0.05％(w/v)、0.04％(w/v)、0.03％(w/v)、0.02％(w/v)或0.01％(w/v)酵母提取物。或者，基本培养基可补充1％(w/v)、0.9％(w/v)、0.8％(w/v)、0.7％(w/v)、0.6％(w/v)、0.5％(w/v)、0.4％(w/v)、0.3％(w/v)、0.2％(w/v)或0.1％(w/v)葡萄糖并补充0.1％(w/v)、0.09％(w/v)、0.08％(w/v)、0.07％(w/v)、0.06％(w/v)、0.05％(w/v)、0.04％(w/v)、0.03％(w/v)或0.02％(w/v)酵母提取物。在一些培养中，可使用显著更高水平的酵母提取物，例如至少1.5％(w/v)、2.0％(w/v)、2.5％(w/v)或3％(w/v)。在(例如酿酒酵母或谷氨酸棒状杆菌的)一些培养中，酵母提取物水平落入以上数值中任意两个的范围内，例如0.5-3.0％(w/v)、1.0-2.5％(w/v)或1.5-2.0％(w/v)。
适用于本文所述的经改造的微生物细胞的维持和生长的示例性材料和方法可参见以下实施例1。
(6E)-8-羟基香叶醇产生和回收
本文所述的任何方法还包括回收(6E)-8-羟基香叶醇步骤。在一些实施方案中，从生产容器回收/收获包含于所谓的收获流(harvest stream)中的所产生的(6E)-8-羟基香叶醇。收获流可包括，例如从生产容器流出的无细胞或含细胞水溶液，其包含由生产容器中的静息细胞(resting cell)转化生产底物而得到的(6E)-8-羟基香叶醇。可通过本领域中已知的任何操作将仍存在于收获流中的细胞与(6E)-8-羟基香叶醇分离，所述操作例如过滤、离心、倾析(decantation)、膜错流超滤或微滤、切向流超滤或微滤或死端过滤。在该细胞分离操作之后，收获流基本上不含细胞。
可任选地进行产生的(6E)-8-羟基香叶醇与收获流中包含的其他组分分离和/或纯化的其他步骤，即所谓的下游处理步骤。这些步骤可包括技术人员已知的任何方式，例如浓缩、萃取、结晶、沉淀、吸附、离子交换和/或色谱法。可单独使用这些步骤中的任何步骤，或将其组合使用以纯化(6E)-8-羟基香叶醇。其他纯化步骤可包括以下方法中的一种或多种：例如，浓缩、结晶、沉淀、清洗和干燥、用活性炭处理、离子交换、纳滤和/或重结晶。适合的纯化方案的设计可取决于细胞、培养基、培养规模、生产容器等，并且在本领域技术水平范围内。
提供以下实施例的目的在于说明公开内容的多种实施方案，并且不意欲以任何方式限制本公开内容。本领域技术人员将能够识别其中的变化和其他用途，其涵盖于本公开内容的宗旨内，由权利要求的范围限定。
实施例1——经改造以产生(6E)-8-羟基香叶醇的酿酒酵母的构建和选择
质粒/DNA设计
本研究中所有测试菌株用质粒DNA转化，所述质粒DNA是使用专用软件设计。质粒设计是特别针对本研究中改造的两种宿主生物之一。通过标准DNA组装方法物理构建质粒DNA。然后，通过分别如下文所述的两种宿主特异性方法之一，将该质粒DNA用于整合代谢途径插入。
酿酒酵母途径整合
“分离标记物，双交换(split-marker，double-crossover)”基因组整合策略被开发以改造酿酒酵母菌株。图2举例说明在酿酒酵母中，互补的分离标记物质粒的基因组整合以及通过菌落PCR确认正确的基因组整合。将具有互补的5’和3’同源臂和URA3选择性标记的重叠部分(以斜线柱表示直接重复)的两种质粒用大范围核酸酶(meganuclease)消化并作为线性片段转化。三重交换事件将所需的异源基因整合到靶基因座并重新构建完整的URA3基因。使用两个3-引物反应来测定源自该整合事件的菌落以确定5’和3’接合(UF/IF/wt-R和DR/IF/wt-F)。对于需要进一步改造的菌株，可将菌株铺在5-FOA平板上以选择URA3去除，留下小的单拷贝的原始直接重复。该基因组整合策略可用于相同操作流程中的基因敲除、基因敲入和启动子滴定(promoter titration)。
细胞培养
为酿酒酵母建立的操作流程包括命中挑选(hit-picking)步骤，该步骤使用在平板上随机化菌株的自动化流程强化成功构建的菌株。对于每个成功构建的菌株，从不同的菌落测试最多达4个重复，以测试菌落之间差异和其他过程差异。如果获得的菌落少于4个，则重复现有菌落，使得从每个需要的基因型测试至少4个孔。
将菌落合并到具有选择性培养基(用于酿酒酵母的SD-ura)的96孔板并培养两天直至饱和，然后用16.6％甘油在-80℃下冷冻保存。然后将冷冻的甘油原液用于在基本培养基中接种种子期，所述基本培养基具有低水平氨基酸以有利于生长和从冷冻中恢复。种子平板在30℃下生长1-2天。然后将种子平板用于接种具有基本培养基的主要培养板并生长48-88小时。在需要的时间点移出平板并测试细胞密度(OD600)、活力和葡萄糖，贮存上清液样品用于目的产物的LC-MS分析。
细胞密度
使用分光光度测定法来测量细胞密度，所述测定检测600nm下各孔的吸光度。机器人技术用于将确定量的培养物从各培养板转移到测定板，然后与175mM磷酸钠(pH 7.0)混合以产生10倍稀释液。使用Tecan M1000分光光度仪测量测定板，并将测定数据上传至LIMS数据库。未接种对照用于减去背景吸光度。通过以下方式来监测细胞生长：在每个时期接种多个平板，然后在每个时间点放弃(sacrifice)整个平板。
为了最小化细胞沉淀同时处理大量平板(这在测量期间可导致非代表性样品)，在每次读数之前振荡每个平板10-15秒。平板内细胞密度的大幅度差异还可导致检测线性范围以外的吸光度测量值，导致对较高OD培养物的低估。总体上，迄今为止测试的菌株的差异尚未显著到足以成为问题的程度。
液体-固体分离
为了收集胞外样品用于LC-MS分析，通过离心分离固相和液相。2000rpm离心培养板4分钟，使用机器人技术将上清液转移到目标板。将75μL上清液转移到各板，其中一个贮存于4℃，第二个贮存于80℃以长期保存。
第一轮基因工程结果
使用酿酒酵母作为宿主细胞，进行第一轮基因工程和筛选。采用文库方法鉴定宿主生物体中的功能性酶。香叶醇合酶序列的广泛检索共鉴定出13个同源序列。在宿主细胞中表达异源香叶醇合酶，在某些情况下，与异源香叶醇-8-羟化酶一起表达。在某些情况下，对香叶醇合酶和/或香叶醇-8-羟化酶核苷酸序列进行密码子优化，用于酿酒酵母。产生菌株并如上所述进行培养，通过LC-MS测量培养基中的(6E)-8-羟基香叶醇效价。菌株和结果示于表1和图2。性能最佳的第一轮菌株为表达紫苏香叶醇合酶以及赤豆香叶醇-8-羟化酶的酿酒酵母菌株，其产生的(6E)-8-羟基香叶醇效价为37.5μg/L培养基。选择该菌株，用于第二轮基因工程和筛选。
Figure BDA0002620930290000291
第二轮基因工程结果
使用组合文库方法将性能最佳的第一轮菌株用作第二轮基因工程的起始宿主。在这一轮中，将1-3种上游途径基因的额外拷贝引入到单独的“子”菌株中，在强的组成型启动子的控制下(表2)。上游途径基因代表在产生(6E)-8-羟基香叶醇的途径中所有参与关键前体(例如乙酰CoA)转化为最终天然代谢物的基因。在组合文库方法中选择在菌株中测试的酶如甲羟戊酸途径图所示(图1)。这一轮中性能最佳的菌株过表达来自酿酒酵母的异戊烯基二磷酸δ-异构酶，得到的(6E)-8-羟基香叶醇的效价为123μg/L培养基。
Figure BDA0002620930290000311
Figure BDA0002620930290000321
Figure BDA0002620930290000331
第三和第四轮基因工程结果
第三轮基因工程没有产生改良菌株，很可能是由于菌株构建错误。第四(提高)轮菌株设计及结果如表3所示。
Figure BDA0002620930290000351
Figure BDA0002620930290000361
Figure BDA0002620930290000371
Figure BDA0002620930290000381
Figure BDA0002620930290000391
Figure BDA0002620930290000401
Figure BDA0002620930290000411
Figure BDA0002620930290000421
Figure BDA0002620930290000431
Figure BDA0002620930290000441
Figure BDA0002620930290000451
Figure BDA0002620930290000461
Figure BDA0002620930290000471
Figure BDA0002620930290000481
实施例2-经改造以产生(6E)-8-羟基香叶醇的解脂亚罗酵母菌株的构建和选择
利用上述用于酿酒酵母的方法，对解脂亚罗酵母进行改造。在第一轮基因工程中构建的菌株及其(6E)-8-羟基香叶醇效价见表4。
Figure BDA0002620930290000501
Figure BDA0002620930290000511
Figure BDA0002620930290000521
Figure BDA0002620930290000531
Figure BDA0002620930290000541
Figure BDA0002620930290000551
Figure BDA0002620930290000561
实施例3——生产(6E)-8-羟基香叶醇的宿主评估
在四种宿主中测试了性能最佳的酶：解脂亚罗酵母、枯草芽孢杆菌、谷氨酸棒状杆菌、酿酒酵母。酿酒酵母的结果如下表5所示。
性能最佳的解脂亚罗酵母菌株生产的效价为310微克/L。该解脂亚罗酵母菌株表达来自紫苏的香叶醇合酶(UniProt ID C0KWV4)、来自赤豆的香叶醇8-羟化酶(UniProt IDA0A0L9UT99)和来自酿酒酵母的异戊烯基二磷酸δ3-δ2-异构酶(UniProt ID P15496)。性能第二佳的解脂亚罗酵母菌株生产200微克/L，并且该菌株表达具有氨基酸置换S80F的大肠杆菌K12的法呢基二磷酸合酶(UniProt ID P22939)[4]、来自葡萄的香叶醇合酶(UniProtID E5GAH8)和来自赤豆的香叶醇8-羟化酶(UniProt ID A0A0L9UT99)。
性能最佳的酿酒酵母菌株生产的效价为217微克/L。该酿酒酵母菌株表达来自紫苏的香叶醇合酶(UniProt ID C0KWV4)、来自赤豆的香叶醇8-羟化酶(UniProt IDA0A0L9UT99)和来自酿酒酵母的异戊烯基二磷酸δ3-δ2-异构酶(UniProt ID P15496)。
枯草芽孢杆菌或谷氨酸棒状杆菌菌株没有产生效价。
Figure BDA0002620930290000581
Figure BDA0002620930290000591
Figure BDA0002620930290000601
Figure BDA0002620930290000611
Figure BDA0002620930290000621
参考文献
1.Polakowski，T.，U.Stahl，and C.Lang，Overexpression of a cytosolichydroxymethylglutaryl-CoA reductase leads to squalene accumulation inyeast.Appl Microbiol Biotechnol，1998.49(1)：p.66-71.
2.Dimster-Denk，D.，M.K.Thorsness，and J.Rine，Feedback regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in Saccharomyces cerevisiae.MolBiol Cell，1994.5(6)：p.655-65.
3.Billingsley，J.M.，et al.，Engineering the biocatalytic selectivity ofiridoid production in Saccharomyces cerevisiae.Metab Eng，2017.44：p.117-125.
4.Eiichiro Ono，N.T.，Method for utilizing monoterpene glycosidationenzyme.2015.
5.Roth，S.，et al.，Chemoenzymatic Synthesis of a Novel Borneol-BasedPolyester.ChemSusChem，2017.10(18)：p.3574-3580.
6.Crowley，J.H.，et al.，A mutation in a purported regulatory geneaffects control of sterol uptake in Saccharomyces cerevisiae.J Bacteriol，1998.180(16)：p.4177-83.
7.Vik，A.and J.Rine，Upc2p and Ecm22p，dual regulators of sterolbiosynthesis in Saccharomyces cerevisiae.Mol Cell Biol，2001.21(19)：p.6395-405.
1.Kanehisa，M.and S.Goto，KEGG：kyoto encyclopedia of genes andgenomes.Nucleic Acids Res，2000.28(1)：p.27-30.
2.Oswald，M.，et al.，Monoterpenoid biosynthesis in Saccharomycescerevisiae.FEMS Yeast Res，2007.7(3)：p.413-21.
3.Polakowski，T.，U.Stahl，and C.Lang，Overexpression of a cytosolichydroxymethylglutaryl-CoA reductase leads to squalene accumulation inyeast.Appl Microbiol Biotechnol，1998.49(1)：p.66-71.
4.Reiling，K.K.，et al.，Mono and diterpene production in Escherichiacoli.Biotechnol Bioeng，2004.87(2)：p.200-12.
序列表
<110> 齐默尔根公司
<120> 通过发酵产生(6E)-8-羟基香叶醇的经改造的生物合成途径
<130> ZMGNP007PUS/ZYM0027-WO-00
<140> Not Assigned
<141> 2018-12-06
<150> US 62/596,013
<151> 2017-12-07
<160> 112
<170> PatentIn version 3.5
<210> 1
<211> 591
<212> PRT
<213> 葡萄（Vitis vinifera）（葡萄）
<400> 1
Met Ser Arg Phe Val Thr Met Pro Ser His Val Leu Pro Ser Ser Phe
1 5 10 15
Val Ala Pro Ser Leu Gln Val Ser Ser Ser Pro Cys Ser Trp Arg Thr
20 25 30
Arg Pro Ser Pro Cys Thr Ser Cys His Leu Ser Pro Ser Ser Ser Ser
35 40 45
Lys Pro Leu Leu Gly Ser His Asp Tyr Ser Leu Leu Lys Ser Leu Thr
50 55 60
Leu Ser Pro His Ala Val Asn Ser Glu Ala Asp Ser Ser Thr Arg Arg
65 70 75 80
Met Lys Glu Val Lys Glu Arg Thr Trp Glu Ala Phe Tyr Arg Ala Trp
85 90 95
Asp Ser Arg Ala Ala Met Glu Met Val Asp Thr Val Glu Arg Leu Gly
100 105 110
Leu Ser Tyr His Phe Glu Asp Glu Ile Asn Ala Leu Leu Gln Arg Phe
115 120 125
Cys Asp Trp Asn Ala Ser Glu Asp Leu Phe Thr Thr Ala Leu Arg Phe
130 135 140
Arg Leu Leu Arg Gln Asn Gly Phe Pro Thr His Ser Asp Val Phe Gly
145 150 155 160
Lys Phe Met Asp Lys Asn Gly Lys Phe Lys Glu Ser Leu Thr Glu Asp
165 170 175
Ile Trp Gly Met Leu Ser Leu His Glu Ala Ser His Leu Gly Ala Lys
180 185 190
Asn Glu Glu Val Leu Ala Glu Ala Lys Glu Phe Thr Arg Ile His Leu
195 200 205
Ile Gln Ser Met Pro His Met Glu Pro His Phe Ser Ser His Val Gly
210 215 220
Arg Ala Leu Glu Leu Pro Arg His Leu Arg Met Val Arg Leu Glu Ala
225 230 235 240
Arg Asn Tyr Ile Gly Glu Tyr Ser Arg Glu Ser Asn Pro Asn Leu Ala
245 250 255
Phe Leu Glu Leu Ala Lys Leu Asp Phe Asp Met Val Gln Ser Leu His
260 265 270
Gln Lys Glu Leu Ala Glu Ile Val Arg Trp Trp Lys Gln Leu Gly Leu
275 280 285
Val Asp Lys Leu Asp Phe Ala Arg Asp Arg Pro Met Glu Cys Phe Leu
290 295 300
Trp Thr Val Gly Ile Phe Pro Asp Pro Arg His Ser Ser Cys Arg Ile
305 310 315 320
Glu Leu Thr Lys Ala Ile Ala Ile Leu Leu Val Ile Asp Asp Ile Tyr
325 330 335
Asp Ser Tyr Gly Ser Leu Asp Glu Leu Ala Leu Phe Thr Asp Ala Val
340 345 350
Lys Arg Trp Asp Leu Gly Ala Met Asp Gln Leu Pro Glu Tyr Met Lys
355 360 365
Ile Cys Tyr Met Ala Leu Tyr Asn Thr Thr Asn Asp Ile Ala Tyr Arg
370 375 380
Ile Leu Lys Glu His Gly Trp Ser Val Ile Glu Asp Leu Lys Arg Thr
385 390 395 400
Trp Met Asp Ile Phe Gly Ala Phe Leu Ala Glu Ala Tyr Cys Phe Lys
405 410 415
Gly Gly His Val Pro Ser Leu Glu Glu Tyr Leu Thr Asn Ala Val Thr
420 425 430
Thr Gly Gly Thr Tyr Met Ala Leu Val His Ala Phe Phe Leu Met Gly
435 440 445
Gln Gly Val Thr Arg Glu Asn Met Ala Met Leu Lys Pro Tyr Pro Asn
450 455 460
Ile Phe Ser Cys Ser Gly Lys Ile Leu Arg Leu Trp Asp Asp Leu Gly
465 470 475 480
Thr Ala Arg Glu Glu Gln Glu Arg Gly Asp Asn Ala Ser Ser Ile Glu
485 490 495
Cys Tyr Lys Arg Glu Arg Glu Met Asp Thr Val Leu Glu Asp Glu Ala
500 505 510
Cys Arg Lys His Ile Arg Gln Met Ile Gln Ser Leu Trp Val Glu Leu
515 520 525
Asn Gly Glu Leu Val Ala Ser Ser Ala Leu Pro Leu Ser Ile Ile Lys
530 535 540
Ala Ala Phe Asn Leu Ser Arg Thr Ala Gln Val Ile Tyr Gln His Gly
545 550 555 560
Asp Asp Asn Lys Thr Ser Ser Val Glu Asp His Val Gln Ala Leu Leu
565 570 575
Phe Arg Pro Val Ser Ser Asn Gly His Ala Gln Ile Thr Met His
580 585 590
<210> 2
<211> 1773
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 2
atgtcgcgtt ttgtcaccat gccctctcac gtcctgcctt catcatttgt cgccccatcc 60
cttcaagttt ccagctcgcc ctgcagctgg cgtacccgac cgtcgccctg caccagctgc 120
cacctttcgc catcgtcgag ctctaaaccg ttgttgggta gccacgatta cagcctgttg 180
aagagcctga ccctttcgcc acacgcggtg aattccgaag cagactcttc gactcgccgt 240
atgaaagaag tgaaagaacg aacctgggag gcattctacc gcgcttggga ttcgcgcgca 300
gcaatggaaa tggtagatac tgttgaacga ctgggacttt cttaccactt cgaagacgaa 360
atcaacgcac tcctccaacg cttttgtgac tggaacgcta gcgaggattt gttcacaact 420
gcgctgcgct ttcgcctgct tcgccaaaac ggtttcccga cccacagcga cgtcttcggc 480
aaatttatgg ataaaaatgg caaattcaaa gaatcactga ccgaagacat ttggggtatg 540
ttgtcccttc acgaggcgtc tcaccttggt gctaaaaacg aagaagtcct tgcggaggct 600
aaagaattta ctcgcatcca tctgattcag tcgatgccac acatggagcc tcatttttcg 660
tcgcacgtgg gccgagctct ggagctgccg cgccacctgc gtatggtccg tttggaagca 720
cgtaactata tcggcgagta ttcgcgcgag tccaatccaa accttgcatt cctggagttg 780
gccaagttgg atttcgatat ggtgcagtct ttgcaccaga aagaattggc cgaaatcgtg 840
cgctggtgga aacagctggg cctcgtggac aagttggatt tcgcacgtga ccgcccaatg 900
gaatgtttcc tctggaccgt tggcatcttt ccggatcccc gacactcctc gtgccgcatc 960
gaacttacca aagctattgc aattctgttg gtgatcgacg acatctatga ttcctacggc 1020
tcgctggatg aactcgctct ctttaccgat gccgtcaagc gctgggatct gggagccatg 1080
gatcaactcc cagaatacat gaagatttgt tacatggcgc tgtacaacac caccaatgat 1140
atcgcttatc gcatcctcaa agagcatggt tggtccgtca tcgaggactt gaaacgaacg 1200
tggatggata tctttggcgc attccttgct gaggcatatt gcttcaaagg aggccatgtg 1260
ccttcgcttg aggaatattt gaccaacgca gttacgactg gcggtaccta tatggcactg 1320
gtgcacgcct tctttcttat gggacagggc gtgacccgcg agaatatggc aatgctgaag 1380
ccctatccta atatctttag ctgctcagga aagatcctcc gtctttggga tgatcttggc 1440
acagcgcgtg aggagcagga gcgtggcgac aacgccagct ccatcgaatg ctacaagcgt 1500
gaacgtgaga tggacaccgt cctcgaagat gaggcatgcc gcaaacacat ccgtcaaatg 1560
attcagtcct tgtgggttga attgaatggc gagcttgtgg cttcatctgc attgccgctg 1620
tccatcatca aggcagcgtt caatctctcc cgcaccgcac aagtgattta tcagcacggt 1680
gacgataaca aaacgtcctc tgttgaggat catgttcaag ccttgctttt ccgtcctgtt 1740
tcctccaatg gtcacgcaca aattaccatg cac 1773
<210> 3
<211> 292
<212> PRT
<213> 葡萄（Vitis vinifera）（葡萄）
<400> 3
Met Glu Cys Phe Leu Trp Thr Val Gly Ile Phe Pro Asp Pro Arg His
1 5 10 15
Ser Ser Cys Arg Ile Glu Leu Thr Lys Ala Ile Ala Ile Leu Leu Val
20 25 30
Ile Asp Asp Ile Tyr Asp Ser Tyr Gly Ser Leu Asp Glu Leu Ala Leu
35 40 45
Phe Thr Asp Ala Val Lys Arg Trp Asp Leu Gly Ala Met Asp Gln Leu
50 55 60
Pro Glu Tyr Met Lys Ile Cys Tyr Met Ala Leu Tyr Asn Thr Thr Asn
65 70 75 80
Asp Ile Ala Tyr Arg Ile Leu Lys Glu His Gly Trp Ser Val Ile Glu
85 90 95
Asp Leu Lys Arg Thr Trp Met Asp Ile Phe Gly Ala Phe Leu Ala Glu
100 105 110
Ala Tyr Cys Phe Lys Gly Gly His Val Pro Ser Leu Glu Glu Tyr Leu
115 120 125
Thr Asn Ala Val Thr Thr Gly Gly Thr Tyr Met Ala Leu Val His Ala
130 135 140
Phe Phe Leu Met Gly Gln Gly Val Thr Arg Glu Asn Met Ala Met Leu
145 150 155 160
Lys Pro Tyr Pro Asn Ile Phe Ser Cys Ser Gly Lys Ile Leu Arg Leu
165 170 175
Trp Asp Asp Leu Gly Thr Ala Arg Glu Glu Gln Glu Arg Gly Asp Asn
180 185 190
Ala Ser Ser Ile Glu Cys Tyr Lys Arg Glu Arg Glu Met Asp Thr Val
195 200 205
Leu Glu Asp Glu Ala Cys Arg Lys His Ile Arg Gln Met Ile Gln Ser
210 215 220
Leu Trp Val Glu Leu Asn Gly Glu Leu Val Ala Ser Ser Ala Leu Pro
225 230 235 240
Leu Ser Ile Ile Lys Ala Ala Phe Asn Leu Ser Arg Thr Ala Gln Val
245 250 255
Ile Tyr Gln His Gly Asp Asp Asn Lys Thr Ser Ser Val Glu Asp His
260 265 270
Val Gln Ala Leu Leu Phe Arg Pro Val Ser Ser Asn Gly His Ala Gln
275 280 285
Ile Thr Met His
290
<210> 4
<211> 876
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 4
atggaatgtt ttctgtggac cgttggtatt ttcccagacc cgcgacactc ttcctgtcgt 60
atcgaactta cgaaagcgat tgcaatcctt ttggtgattg atgatatcta cgactcgtac 120
ggctcattgg acgaacttgc gctcttcacc gatgcagtaa aacgctggga tttgggcgcc 180
atggatcaat tgccggaata catgaagatc tgctatatgg cattgtacaa caccacgaac 240
gacatcgcat atcgtatcct caaagagcac ggttggtctg taatcgaaga cctcaagcgc 300
acctggatgg atatcttcgg agcgttcctc gcagaagcgt attgcttcaa gggtggacac 360
gttccttccc tcgaagaata cctgacaaac gcggttacta ccggaggtac atacatggct 420
ctcgtgcatg ctttctttct tatgggtcag ggtgttaccc gcgaaaacat ggcgatgctt 480
aaaccgtatc caaacatctt ctcgtgttca ggcaagatcc tgcgcttgtg ggatgatttg 540
ggaacagctc gcgaagagca ggagcgtggt gataacgcaa gctctatcga atgttacaaa 600
cgtgaacgcg aaatggacac agtcctggag gatgaagcct gccgcaagca cattcgccag 660
atgatccaat ccctctgggt tgaactgaac ggcgaactcg ttgcaagctc cgcgctgccg 720
ctttcgatca ttaaggcggc cttcaatctc tcgcgcactg cacaggttat ttaccagcat 780
ggcgatgaca ataaaacgtc ctccgtagaa gatcatgtgc aggctctgtt gtttcgccca 840
gtgagcagca acggacacgc acagatcacg atgcac 876
<210> 5
<211> 603
<212> PRT
<213> 紫苏（Perilla setoyensis）
<400> 5
Met Cys Ser Ile Ser Gln Lys Val Val Ile Gly Leu Asn Lys Ala Ala
1 5 10 15
Ala Asn Asn Cys Leu Gln Asn Leu Asp Arg Arg Gly Phe Lys Thr Arg
20 25 30
Arg Val Ser Ser Ser Glu Ala Ala Ser Cys Leu Arg Ala Ser Ser Ser
35 40 45
Leu Gln Leu Asp Val Lys Pro Val Glu Glu Gly Arg Arg Ser Gly Asn
50 55 60
Tyr Gln Pro Ser Ile Trp Asp Phe Asn Tyr Val Gln Ser Leu Asn Thr
65 70 75 80
Pro Tyr Lys Glu Glu Arg Tyr Leu Thr Arg His Ala Glu Leu Ile Val
85 90 95
Gln Val Lys Pro Leu Leu Glu Lys Lys Met Glu Ala Thr Gln Gln Leu
100 105 110
Glu Leu Ile Asp Asp Leu Asn Asn Leu Gly Leu Ser Tyr Phe Phe Gln
115 120 125
Asp Arg Ile Lys Gln Ile Leu Ser Phe Ile Tyr Asp Glu Asn Gln Cys
130 135 140
Phe His Ser Asn Ile Asn Asp Gln Ala Glu Lys Arg Asp Leu Tyr Phe
145 150 155 160
Thr Ala Leu Gly Phe Arg Leu Leu Arg Gln His Gly Phe Asn Val Ser
165 170 175
Gln Glu Val Phe Asp Cys Phe Lys Asn Asp Lys Gly Ser Asp Phe Lys
180 185 190
Ala Ser Leu Ser Gly Asn Thr Lys Gly Leu Leu Gln Leu Tyr Glu Ala
195 200 205
Ser Phe Leu Val Arg Glu Gly Glu Asp Thr Leu Glu Leu Ala Arg Gln
210 215 220
Phe Ala Thr Lys Phe Leu Arg Arg Lys Leu Asp Glu Ile Asp Asp Asn
225 230 235 240
His Leu Leu Ser Arg Ile His His Ser Leu Glu Ile Pro Leu His Trp
245 250 255
Arg Ile Gln Arg Leu Glu Ala Arg Trp Phe Leu Asp Ala Tyr Ala Thr
260 265 270
Arg His Asp Met Asn Pro Ile Ile Leu Glu Leu Ala Lys Leu Asp Phe
275 280 285
Asn Ile Ile Gln Ala Thr His Gln Glu Glu Leu Lys Asp Val Ser Arg
290 295 300
Trp Trp Gln Asn Thr Arg Leu Ala Glu Lys Leu Pro Phe Val Arg Asp
305 310 315 320
Arg Leu Val Glu Ser Tyr Phe Trp Ala Ile Ala Leu Phe Glu Pro His
325 330 335
Gln Tyr Gly Tyr Gln Arg Arg Val Ala Ala Lys Ile Ile Thr Leu Ala
340 345 350
Thr Ser Ile Asp Asp Val Tyr Asp Ile Tyr Gly Thr Leu Asp Glu Leu
355 360 365
Gln Leu Phe Thr Asp Asn Phe Arg Arg Trp Asp Thr Glu Ser Leu Gly
370 375 380
Gly Leu Pro Tyr Ser Met Gln Leu Phe Tyr Met Val Ile His Asn Phe
385 390 395 400
Val Ser Glu Leu Ala Tyr Glu Ile Leu Lys Glu Lys Gly Phe Ile Ala
405 410 415
Ile Pro Tyr Leu Gln Arg Ser Trp Val Asp Leu Ala Glu Ser Phe Leu
420 425 430
Lys Glu Ala Asn Trp Tyr Tyr Ser Gly Tyr Thr Pro Ser Leu Glu Glu
435 440 445
Tyr Ile Asp Asn Gly Ser Ile Ser Ile Gly Ala Val Ala Val Leu Ser
450 455 460
Gln Val Tyr Phe Thr Leu Ala Asn Ser Ile Glu Lys Pro Lys Ile Glu
465 470 475 480
Ser Met Tyr Lys Tyr His His Ile Leu Arg Leu Ser Gly Leu Leu Val
485 490 495
Arg Leu His Asp Asp Leu Gly Thr Ser Leu Phe Glu Lys Lys Arg Gly
500 505 510
Asp Val Pro Lys Ala Val Glu Ile Cys Met Lys Glu Arg Asn Asp Thr
515 520 525
Glu Glu Glu Ala Glu Glu His Val Lys Tyr Leu Ile Arg Glu Ala Trp
530 535 540
Lys Glu Met Asn Thr Ala Thr Ala Ala Ala Gly Cys Pro Phe Met Asp
545 550 555 560
Glu Leu Asn Val Ala Ala Ala Asn Leu Gly Arg Ala Ala Gln Phe Val
565 570 575
Tyr Leu Asp Gly Asp Gly His Gly Val Gln His Ser Lys Ile His Gln
580 585 590
Gln Met Gly Gly Leu Met Phe Lys Pro Tyr Val
595 600
<210> 6
<211> 1809
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 6
atgtgcagca tctctcaaaa ggtcgttatt ggattgaata aggcagcggc caataactgc 60
ctccaaaacc ttgatcgccg aggttttaaa acacgccgtg tttcatcctc cgaagcggct 120
tcttgcctcc gagcctcaag ctcgctgcag cttgatgtca agccggtgga ggaaggccga 180
cgttccggta actatcagcc atcaatctgg gattttaatt atgtccagtc cctcaacacc 240
ccctacaaag aggaacgtta ccttacccga cacgcggaac tcatcgttca ggtcaagccg 300
cttctcgaaa aaaagatgga ggctactcag cagttggagc tcatcgatga ccttaacaac 360
ctcggtctga gctacttttt ccaagaccgc atcaagcaga ttctctcatt catctatgat 420
gaaaatcagt gtttccactc caatatcaat gatcaagctg aaaagcgaga cctgtacttt 480
accgcccttg gatttcgtct ccttcgccag cacggcttta acgtgtctca ggaggtgttc 540
gattgcttca aaaacgataa aggctccgat tttaaagcgt ccctgtctgg aaacaccaag 600
ggcctcctcc agctctacga agcctccttc ttggtccgtg agggtgaaga caccctcgaa 660
ctggcccgtc agttcgcgac caagttcctg cgccgcaagc ttgacgaaat cgacgacaac 720
catctgctct cacgaattca ccactcactc gaaatcccgt tgcactggcg catccagcgt 780
ctggaggcgc gctggtttct ggacgcgtac gctacacgcc acgatatgaa tcccattatc 840
ttggagctcg caaaactgga ctttaatatt attcaagcaa cccaccaaga agagctcaag 900
gacgtgagcc gttggtggca gaatacgcgc ctggccgaaa agctcccgtt tgtgcgcgat 960
cgacttgtcg agtcatactt ttgggccatc gcactctttg agccacatca atatggatac 1020
cagcgccgtg ttgctgccaa gattatcacc ctcgcgacat ccatcgatga cgtgtacgac 1080
atctacggca ctctcgacga gctccaactt tttaccgata acttccgtcg ctgggacacg 1140
gaatccttgg gcggtcttcc ttactcgatg cagttgttct acatggtgat tcacaacttt 1200
gtctctgagt tggcctatga aatcctgaaa gaaaaaggtt ttatcgccat cccctacctc 1260
caacgctcct gggttgattt ggcagaatcc tttttgaaag aggcaaactg gtactactct 1320
ggttatacac cctccctcga agaatacatc gacaacggct ccatctcaat tggcgcagta 1380
gctgtgttgt cccaagttta tttcactttg gcaaattcta tcgaaaagcc aaagattgaa 1440
tctatgtaca agtaccacca tattttgcgt ctgtcgggtt tgttggtgcg cctgcacgat 1500
gacttgggaa cttccttgtt cgagaagaag cgcggcgatg ttcctaaggc ggtcgaaatt 1560
tgcatgaaag aacgcaacga tacggaggag gaagcagaag aacacgttaa gtacttgatt 1620
cgcgaagcct ggaaggagat gaacactgct acggcagctg caggatgtcc ttttatggat 1680
gagctcaacg tggctgctgc caatctgggt cgcgcagccc agttcgtgta cctcgacgga 1740
gacggtcatg gcgtgcagca ctccaaaatc catcaacaga tgggtggcct tatgtttaaa 1800
ccgtatgtg 1809
<210> 7
<211> 495
<212> PRT
<213> 川西獐牙菜（Swertia mussotii）（龙胆科植物（Felwort））
<400> 7
Met Asp Phe Asp Phe Leu Thr Ile Ala Ile Gly Phe Leu Phe Thr Ile
1 5 10 15
Thr Leu Tyr Gln Ala Leu Asn Phe Phe Ser Arg Lys Ser Lys Asn Leu
20 25 30
Pro Pro Gly Pro Ser Pro Leu Pro Leu Ile Gly Asn Leu His Leu Leu
35 40 45
Gly Asp Gln Pro His Lys Ser Leu Ala Lys Leu Ala Lys Lys His Gly
50 55 60
Pro Ile Met Gly Leu Gln Leu Gly Gln Val Thr Thr Ile Val Val Thr
65 70 75 80
Ser Ser Gly Met Ala Lys Glu Val Leu Gln Lys Gln Asp Leu Ala Phe
85 90 95
Ser Ser Arg Ser Ile Pro Asn Ala Ile His Ala His Asp Gln Tyr Lys
100 105 110
Tyr Ser Val Ile Trp Leu Pro Val Ala Ser Arg Trp Arg Gly Leu Arg
115 120 125
Lys Ala Leu Asn Ser Asn Met Phe Ser Gly Asn Arg Leu Asp Ala Asn
130 135 140
Gln His Leu Arg Ser Arg Lys Val Gln Glu Leu Ile Ala Tyr Cys Arg
145 150 155 160
Lys Ser Ser Gln Thr Gly Asp Ala Ile Asp Val Gly Arg Ala Ala Phe
165 170 175
Arg Thr Ser Leu Asn Leu Leu Ser Asn Thr Met Phe Ser Lys Asp Leu
180 185 190
Thr Asp Pro Tyr Ser Asp Ser Ala Lys Glu Phe Lys Asp Leu Val Trp
195 200 205
Asn Val Met Val Glu Ala Gly Lys Pro Asn Leu Val Asp Tyr Phe Pro
210 215 220
Leu Leu Asp Lys Val Asp Pro Gln Gly Ile Arg Lys Arg Met Thr Ile
225 230 235 240
His Phe Gly Lys Ile Leu Glu Leu Phe Gly Gly Leu Ile Asp Glu Arg
245 250 255
Leu Gln Gln Lys Lys Ala Lys Gly Val Asn Asp Asp Val Leu Asp Val
260 265 270
Leu Leu Thr Thr Ser Glu Glu Ser Pro Glu Glu Ile Asp Arg Thr His
275 280 285
Ile Gln Arg Met Cys Leu Asp Leu Phe Val Ala Gly Thr Asp Thr Thr
290 295 300
Ser Ser Thr Leu Glu Trp Ala Met Ser Glu Met Leu Lys Asn Pro Glu
305 310 315 320
Lys Met Lys Ala Ala Gln Ala Glu Leu Ala Gln Val Ile Gly Lys Gly
325 330 335
Lys Ala Val Glu Glu Ala Asp Leu Ala Arg Leu Pro Tyr Leu Arg Cys
340 345 350
Ala Ile Lys Glu Thr Leu Arg Ile His Pro Pro Val Pro Leu Leu Ile
355 360 365
Pro Arg Arg Thr Glu Gln Glu Val Glu Val Cys Gly Tyr Thr Val Pro
370 375 380
Lys Asn Ser Gln Val Leu Val Asn Val Trp Ala Ile Ser Arg Asp Asp
385 390 395 400
Ala Ile Trp Lys Asp Pro Leu Ser Phe Lys Pro Glu Arg Phe Leu Glu
405 410 415
Ser Glu Leu Glu Met Arg Gly Lys Asp Phe Glu Leu Ile Pro Phe Gly
420 425 430
Ala Gly Arg Arg Ile Cys Pro Gly Leu Pro Leu Ala Val Arg Met Val
435 440 445
Pro Val Met Leu Gly Ser Leu Leu Asn Ser Phe Asp Trp Lys Leu Glu
450 455 460
Gly Gly Ile Ala Pro Lys Asp Leu Asp Met Glu Glu Lys Phe Gly Ile
465 470 475 480
Thr Leu Gln Lys Ala His Pro Leu Arg Ala Val Ala Thr Pro Leu
485 490 495
<210> 8
<211> 1486
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 8
atggatttcg acttccttac tatcgcgatt ggttttctgt ttactatcac tttgtaccaa 60
gcgcttaact tcttttcccg taaatcaaag aatctcccac caggtccgtc cccattgcca 120
ctcattggta acttgcatct gctcggtgac caaccccata agtcactggc caaactcgca 180
aagaaacatg gtcctatcat gggactgcaa ctgggtcaag tcaccaccat cgttgtaacg 240
tcttctggta tggcgaagga ggttctccag aagcaggacc tggccttttc aagccgttcc 300
atcccaaacg caatccacgc ccatgaccaa tataagtata gcgtgatttg gctcccggtg 360
gcgtcccgat ggcgaggtct gcgtaaggcg ctcaactcca atatgttctc tggaaatcgc 420
ctggatgcaa atcagcactt gcgttcacga aaggtccagg aactgatcgc ctattgccgc 480
aagtcatctc aaactggcga tgctattgat gttggtcgcg cagcattccg cacttcattg 540
aacctgctta gcaacaccat gttcagcaag gaccttacag acccttattc tgactccgca 600
aaagaattta aggatctcgt ttggaacgtg atggtggagg caggcaaacc caatttggtt 660
gactacttcc ctctgttgga taaagttgac ccccaaggca ttcgaaagcg catgaccatt 720
catttcggta aaatcctcga attgtttggc ggacttatcg acgaacgatt gcagcaaaaa 780
aaagcaaagg gagtgaacga cgacgtgctg gacgtgctcc ttaccacttc agaggaaagc 840
ccggaggaaa tcgaccgtac ccatatccag cgcatgtgcc tggacttgtt cgtcgcgggc 900
acggacacta cgtcgtcgac gctcgaatgg gccatgtctg aaatgcttaa gaaccccgaa 960
aaaatgaagg cggcccaggc agaactggcc caggttattg gaaaaggcaa ggctgttgaa 1020
gaagcagact tggcgcgcct gccatacctg cgctgcgcta ttaaagagac ccttcgaatc 1080
catcctccgg ttccactgct gattcctcga cgcacggaac aagaggtcga ggtgtgcggt 1140
tatacggttc cgaagaactc acaggttttg gtgaacgttt gggccatctc tcgagacgat 1200
gcaatttgga aggatccact ctcctttaag cctgaacgtt ttctggagtc tgaattggag 1260
atgcgtggta aggatttcga gcttatccca ttcggtgcgg gccgacgcat ttgcccgggc 1320
cttcctctgg cggtgcgcat ggtcccagtg atgcttggat ctcttttgaa ctcattcgat 1380
tggaagctgg agggcggcat cgcgcctaag gacttggaca tggaagaaaa gttcggcatc 1440
acgctgcaga aagcccaccc tctccgcgct gtcgccactc cactgt 1486
<210> 9
<211> 505
<212> PRT
<213> 毛果杨（Populus trichocarpa）（毛果杨（Western balsam poplar））（香脂杨亚种毛果杨（Populus balsamifera subsp. Trichocarpa））
<400> 9
Met Asn Phe Phe Ile Ser Val Leu Leu Tyr Phe Leu Leu Thr Phe Ala
1 5 10 15
Val Ile Gln Ser Leu Asp Tyr Ile Leu Arg Arg Ser Lys Arg Lys Ser
20 25 30
Gly Lys Leu Pro Pro Gly Pro Ser Arg Leu Pro Ile Val Gly Asn Leu
35 40 45
Leu Asp Leu Gly Asp Lys Pro His Lys Ser Leu Ala Lys Leu Ala Lys
50 55 60
Thr His Gly Gln Leu Met Ser Leu Lys Leu Gly Gln Val Thr Thr Ile
65 70 75 80
Val Val Ser Ser Ala Thr Met Ala Lys Glu Val Leu Gln Lys His Asp
85 90 95
Leu Thr Phe Cys Asn Arg Thr Val Val Asp Ala Val Arg Ala Leu Asp
100 105 110
His His Glu Ala Gly Ile Ala Trp Leu Pro Val Ala Thr Arg Trp Arg
115 120 125
Asn Leu Arg Lys Ile Cys Asn Ser His Ile Phe Thr Ser Gln Lys Leu
130 135 140
Asp Ala Asn Gln Asp Leu Arg Arg Lys Lys Val Gln Asp Leu Leu Ala
145 150 155 160
Glu Val Gln Glu Arg Cys Leu Val Gly Glu Ala Val Asp Leu Arg Gln
165 170 175
Ala Ala Phe Thr Ala Thr Leu Asn Ala Leu Ser Asn Thr Val Leu Ser
180 185 190
Leu Asp Leu Thr Asp Leu Ser Ser Asp Ile Ala Arg Glu Phe Lys Glu
195 200 205
His Ile Ser Cys Ile Met Asp Glu Ala Gly Lys Pro Asn Leu Val Asp
210 215 220
Tyr Phe Pro Leu Leu Arg Arg Ile Asp Pro Gln Gly Ile Arg Arg Arg
225 230 235 240
Thr Ala Ile His Phe Gly Lys Val Phe Asp Leu Phe Asp Arg Leu Ile
245 250 255
Ile Glu Arg Leu Gln Leu Arg Lys Val Lys Gly Tyr Ile Pro Leu Asp
260 265 270
Asp Met Leu Asp Thr Leu Leu Thr Ile Ser Glu Val Asn Asn Glu Glu
275 280 285
Met Asp Ala Thr Arg Ile Lys His Phe Phe Leu Asp Leu Phe Gly Ala
290 295 300
Gly Thr Asp Thr Thr Ser Ser Thr Leu Glu Trp Ala Met Ala Glu Leu
305 310 315 320
Leu His Ser Pro Lys Thr Leu Leu Lys Ala Arg Ala Glu Leu Glu Arg
325 330 335
Thr Ile Gly Glu Gly Asn Leu Leu Glu Glu Ser Asp Ile Thr Arg Leu
340 345 350
Pro Tyr Leu Gln Ala Val Ile Lys Glu Thr Leu Arg Leu His Pro Ala
355 360 365
Val Pro Phe Leu Leu Pro His Lys Ala Gly Ala Asp Ala Glu Ile Gly
370 375 380
Gly Phe Thr Val Pro Lys Asn Ala Gln Val Leu Val Asn Val Trp Ala
385 390 395 400
Ile Gly Arg Asp Pro Ser Met Trp Glu Asp Pro Asn Ser Phe Val Pro
405 410 415
Glu Arg Phe Leu Glu Ser Gly Ile Asp His Arg Gly Gln Asn Phe Glu
420 425 430
Phe Ile Pro Phe Gly Ser Gly Arg Arg Ile Cys Pro Gly Leu Pro Leu
435 440 445
Ala Met Arg Met Leu Pro Leu Met Leu Gly Ser Leu Ile Leu Ser Phe
450 455 460
Asp Trp Lys Leu Ala Asp Gly Val Thr Pro Glu Asn Leu Asn Met Asp
465 470 475 480
Asp Lys Phe Gly Leu Thr Leu Leu Lys Ala Gln Pro Leu Arg Ala Ile
485 490 495
Pro Ile Thr Arg Glu Leu Lys His Gly
500 505
<210> 10
<211> 1515
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 10
atgaactttt tcatctctgt gctgttgtat ttccttctga cattcgcggt gattcaatcc 60
ctggactaca tccttcgccg ctccaagcgt aagtccggca aactgccgcc aggcccttca 120
cgacttccga ttgttggaaa cctgctggac ctgggagaca agccccacaa atccctcgca 180
aagctggcga aaacccacgg ccaattgatg tcacttaagc tcggacaagt gaccacgatc 240
gtggtttcaa gcgctaccat ggccaaggag gtgttgcaga agcatgatct gactttctgt 300
aaccgcactg ttgttgacgc tgttcgcgcg ctggaccacc atgaggccgg cattgcgtgg 360
ctgccagtgg ctacccgttg gcgcaacctg cgtaaaattt gcaatagcca tatctttaca 420
tcccagaaac tcgacgctaa tcaagacctg cgacgcaaaa aagtccagga tctgctggca 480
gaagtgcaag agcgctgcct ggtaggtgag gctgttgact tgcgtcaagc agcatttacc 540
gcgactctta atgccctgag caacactgtt ttgtcgctcg acctcaccga tctgtcatca 600
gatatcgctc gcgagtttaa agagcacatc tcctgcatta tggatgaagc gggcaagcca 660
aacctggttg attactttcc tttgcttcgc cgcatcgacc cgcagggcat tcgtcgccgc 720
actgcaattc acttcggcaa ggtgttcgat ctcttcgatc gcttgattat cgaacgtttg 780
cagctccgca aagtcaaggg ttacattccc ctggacgaca tgctggacac tttgctcacg 840
atctcagaag tcaacaacga agagatggac gctacacgta tcaagcactt ttttttggat 900
ttgttcggcg caggcaccga tacgacttca tcgactcttg agtgggccat ggcagagctc 960
ttgcactcgc cgaaaaccct cctcaaagcc cgcgctgaac tggaacgcac catcggcgag 1020
ggcaatcttt tggaggaatc cgacatcacc cgtctcccct acctgcaggc tgtcatcaag 1080
gagactctcc gactccatcc agccgtgccc ttcctccttc cacacaaggc aggcgctgat 1140
gctgaaatcg gcggttttac tgtgccaaaa aacgcgcagg tcttggtgaa tgtctgggcg 1200
attggccgtg atccctcaat gtgggaagac cccaattcct tcgtccctga acgcttcttg 1260
gaatcaggta tcgatcaccg aggacaaaac tttgaattta tccctttcgg cagcggacgt 1320
cgaatttgcc caggtcttcc gctggcgatg cgtatgctcc ctctcatgct cggttctctc 1380
atcttgtcct tcgactggaa gttggcagat ggtgtgacgc cagaaaactt gaacatggac 1440
gataagttcg gtttgactct cctgaaagct cagccccttc gtgcgatccc catcacccga 1500
gaattgaaac acggt 1515
<210> 11
<211> 501
<212> PRT
<213> 赤豆（Phaseolus angularis）（红豆（Azuki bean））（赤豆（Vigna angularis））
<400> 11
Met Glu Thr Asn Ser Thr Leu Phe Phe Thr Phe Thr Val Leu Leu Ile
1 5 10 15
Thr Ile Ile Thr Phe Leu Lys Ala Leu Lys Ser Leu Phe Thr Pro Asn
20 25 30
Lys Ser Lys Ser Asn Leu Pro Pro Gly Pro Lys Gly Leu Pro Leu Val
35 40 45
Gly Asn Leu Leu Gln Leu Gly Ala Lys Pro His Gln Thr Leu Ala Thr
50 55 60
Leu Ala Asn Ile His Gly Ser Ile Met Ser Leu Lys Leu Gly Gln Glu
65 70 75 80
Thr Thr Ile Val Met Ser Ser Ala Glu Ala Ala Lys Gly Val Leu Gln
85 90 95
Ile His Asp His Phe Leu Ser Asn Arg Lys Ile Pro Asp Ala Met Arg
100 105 110
Gly Ser Ser His Asp His Phe Ser Leu Pro Phe Met Pro Val Ser Gln
115 120 125
Gln Trp Arg Glu Leu Arg Lys Leu Cys Asn Glu Leu Leu Phe Ser Asn
130 135 140
Lys Asn Leu Asp Ala Thr Gln Gly Leu Arg Ser Lys Lys Val Arg Glu
145 150 155 160
Leu Tyr Ser Asp Ile His Arg Ser Ser Leu Lys Gly Glu Pro Val Asn
165 170 175
Ile Gly Arg Leu Ala Phe Lys Thr Thr Ile Asn Gln Leu Ser Asn Thr
180 185 190
Ile Tyr Ser Glu Asp Phe Leu Gln Ser Ala Glu Lys Ala Gly Glu Met
195 200 205
Lys Glu Leu Val Thr Asn Ile Met Lys Glu Val Gly Arg Pro Asn Leu
210 215 220
Ala Asp Cys Phe Pro Val Leu Lys Met Ile Asp Pro His Gly Ile Arg
225 230 235 240
Arg Arg Thr Gly Ser Tyr Phe Ser Lys Leu Leu Asn Ile Phe Lys Ser
245 250 255
Leu Ile His Lys Arg Leu Glu Leu Arg Lys Asp Ala Ala Gly Tyr Cys
260 265 270
Thr Lys Lys Asp Met Leu Asp Ala Met Leu Asn Asp Ala Gln His Lys
275 280 285
Met Asp Ile Val Lys Ile Gln Arg Leu Ser Leu Asp Leu Phe Val Ala
290 295 300
Gly Thr Asp Thr Val Thr Ser Thr Val Glu Trp Ala Met Ala Glu Leu
305 310 315 320
Leu His Asn Pro His Val Met Ser Lys Ala Lys Glu Glu Leu Glu Arg
325 330 335
Ile Ile Gly Lys Asp Asn Leu Val Glu Glu Ser Asp Ile Ala Lys Leu
340 345 350
Pro Tyr Leu Gln Ala Ile Val Lys Glu Thr Phe Arg Leu His Pro Ala
355 360 365
Val Pro Leu Leu Leu Pro Arg Lys Ala Glu Val Glu Phe Glu Met His
370 375 380
Gly Tyr Thr Ile Pro Lys Gly Ala Gln Val Leu Ile Asn Val Trp Ala
385 390 395 400
Ile Gly Arg Asp Pro Asn Leu Trp Glu Lys Pro Arg Leu Phe Trp Pro
405 410 415
Glu Arg Phe Leu Glu Ser Glu Ile Asp Phe Lys Gly Arg Ser Phe Glu
420 425 430
Leu Thr Pro Phe Gly Gly Gly Arg Arg Ile Cys Pro Gly Leu Pro Leu
435 440 445
Ala Ile Arg Leu Val Phe Leu Met Leu Gly Leu Phe Ile Asn Ser Phe
450 455 460
Asp Trp Glu Leu Gln Asp Ile Gln Pro Glu Asp Met Asn Met Asp Glu
465 470 475 480
Asn Phe Gly Leu Thr Leu Glu Lys Ala Gln Pro Val Leu Ala Ile Pro
485 490 495
Ile Ile Pro Lys His
500
<210> 12
<211> 1503
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 12
atggagacta actctaccct tttctttaca ttcaccgtcc tcctgatcac aatcattacg 60
ttcctgaagg cgttgaaatc tttgtttacc cctaacaagt caaaatctaa cctcccaccc 120
ggtcccaagg gccttccttt ggttggcaac ctgcttcagt tgggcgcgaa gccgcaccag 180
accctcgcaa ccctcgcaaa catccacgga tccatcatgt ccctcaaatt gggtcaagag 240
acaaccatcg tgatgtcttc cgctgaggcc gcaaagggag ttctgcagat tcatgatcat 300
ttcctctcaa accgcaaaat ccccgacgca atgcgcggct cctcccacga ccacttctcc 360
cttcctttca tgcctgtttc ccaacagtgg cgagaattgc gaaagctttg caacgaactc 420
ctgttctcca acaagaacct ggacgctaca caaggcctgc gctccaaaaa ggttcgagaa 480
ctgtacagcg atattcaccg cagctcgctg aagggcgagc ccgtgaacat cggtcgtctg 540
gcttttaaaa cgaccatcaa ccaactttct aacaccatct actcggaaga cttcttgcaa 600
tccgccgaga aagcgggcga aatgaaggaa ctcgttacca atattatgaa ggaggttggc 660
cgcccgaacc ttgccgactg cttccccgtt ctgaaaatga tcgaccccca cggcatccgc 720
cgtcgaaccg gctcttactt cagcaagctg cttaatatct tcaaatccct gatccacaag 780
cgtctggaac ttcgtaaaga tgctgcgggc tactgtacta aaaaggatat gcttgacgcg 840
atgctgaacg acgcccaaca caaaatggac atcgttaaga tccagcgtct ctctcttgat 900
ttgttcgttg caggaacgga taccgtcacc tccactgtcg agtgggctat ggctgaactg 960
cttcacaacc cccacgttat gtcaaaggct aaagaggaac tcgaacgcat tatcggtaag 1020
gataatctgg tggaggagtc agacatcgcc aagttgcctt acttgcaggc tatcgtgaag 1080
gagaccttcc gtctccaccc agcagtcccc cttttgctgc cacgcaaagc cgaggttgaa 1140
tttgaaatgc acggttacac aatccccaaa ggcgcccagg tcctgatcaa tgtctgggcc 1200
attggtcgcg acccgaactt gtgggaaaag ccacgcctgt tttggcccga gcgatttctt 1260
gaatccgaga tcgacttcaa gggccgatcg ttcgagctca ccccgttcgg cggcggccgt 1320
cgcatctgcc ctggactccc attggcgatc cgacttgttt ttctgatgct cggcctgttt 1380
attaactctt tcgattggga acttcaggac attcagccgg aagacatgaa tatggacgaa 1440
aacttcggtc tgactcttga aaaggcccag ccagttctcg caattccaat cattccaaaa 1500
cac 1503
<210> 13
<211> 443
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 13
Met Ser Leu Pro Phe Leu Thr Ser Ala Pro Gly Lys Val Ile Ile Phe
1 5 10 15
Gly Glu His Ser Ala Val Tyr Asn Lys Pro Ala Val Ala Ala Ser Val
20 25 30
Ser Ala Leu Arg Thr Tyr Leu Leu Ile Ser Glu Ser Ser Ala Pro Asp
35 40 45
Thr Ile Glu Leu Asp Phe Pro Asp Ile Ser Phe Asn His Lys Trp Ser
50 55 60
Ile Asn Asp Phe Asn Ala Ile Thr Glu Asp Gln Val Asn Ser Gln Lys
65 70 75 80
Leu Ala Lys Ala Gln Gln Ala Thr Asp Gly Leu Ser Gln Glu Leu Val
85 90 95
Ser Leu Leu Asp Pro Leu Leu Ala Gln Leu Ser Glu Ser Phe His Tyr
100 105 110
His Ala Ala Phe Cys Phe Leu Tyr Met Phe Val Cys Leu Cys Pro His
115 120 125
Ala Lys Asn Ile Lys Phe Ser Leu Lys Ser Thr Leu Pro Ile Gly Ala
130 135 140
Gly Leu Gly Ser Ser Ala Ser Ile Ser Val Ser Leu Ala Leu Ala Met
145 150 155 160
Ala Tyr Leu Gly Gly Leu Ile Gly Ser Asn Asp Leu Glu Lys Leu Ser
165 170 175
Glu Asn Asp Lys His Ile Val Asn Gln Trp Ala Phe Ile Gly Glu Lys
180 185 190
Cys Ile His Gly Thr Pro Ser Gly Ile Asp Asn Ala Val Ala Thr Tyr
195 200 205
Gly Asn Ala Leu Leu Phe Glu Lys Asp Ser His Asn Gly Thr Ile Asn
210 215 220
Thr Asn Asn Phe Lys Phe Leu Asp Asp Phe Pro Ala Ile Pro Met Ile
225 230 235 240
Leu Thr Tyr Thr Arg Ile Pro Arg Ser Thr Lys Asp Leu Val Ala Arg
245 250 255
Val Arg Val Leu Val Thr Glu Lys Phe Pro Glu Val Met Lys Pro Ile
260 265 270
Leu Asp Ala Met Gly Glu Cys Ala Leu Gln Gly Leu Glu Ile Met Thr
275 280 285
Lys Leu Ser Lys Cys Lys Gly Thr Asp Asp Glu Ala Val Glu Thr Asn
290 295 300
Asn Glu Leu Tyr Glu Gln Leu Leu Glu Leu Ile Arg Ile Asn His Gly
305 310 315 320
Leu Leu Val Ser Ile Gly Val Ser His Pro Gly Leu Glu Leu Ile Lys
325 330 335
Asn Leu Ser Asp Asp Leu Arg Ile Gly Ser Thr Lys Leu Thr Gly Ala
340 345 350
Gly Gly Gly Gly Cys Ser Leu Thr Leu Leu Arg Arg Asp Ile Thr Gln
355 360 365
Glu Gln Ile Asp Ser Phe Lys Lys Lys Leu Gln Asp Asp Phe Ser Tyr
370 375 380
Glu Thr Phe Glu Thr Asp Leu Gly Gly Thr Gly Cys Cys Leu Leu Ser
385 390 395 400
Ala Lys Asn Leu Asn Lys Asp Leu Lys Ile Lys Ser Leu Val Phe Gln
405 410 415
Leu Phe Glu Asn Lys Thr Thr Thr Lys Gln Gln Ile Asp Asp Leu Leu
420 425 430
Leu Pro Gly Asn Thr Asn Leu Pro Trp Thr Ser
435 440
<210> 14
<211> 1329
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 14
atgtccctgc cattcctcac gtccgcacca ggaaaagtca ttatctttgg cgaacattcc 60
gcagtgtaca acaagcctgc cgtggctgcc tccgtgagcg ccctccgcac ctacttgctg 120
attagcgaga gctctgcccc agataccatc gagctggatt ttccagacat ctcctttaac 180
cataaatggt ccattaacga ttttaacgcg attaccgaag atcaagtcaa cagccagaag 240
ctcgcaaagg cacaacaggc aacagatggc ctctcccagg agcttgtttc cctcctggat 300
ccactgcttg cacagctctc agaatccttt cactaccatg ccgctttctg tttcctgtac 360
atgttcgtgt gtctctgccc ccacgcgaag aacatcaagt tcagcctgaa gtcgacactc 420
cctatcggtg ctggtctcgg atcgtccgcg agcatctcgg tctcgttggc acttgcaatg 480
gcatacctcg gtggtcttat tggttcaaac gatctggaaa agttgtcgga aaacgataaa 540
catattgtga accagtgggc attcatcggc gaaaaatgta tccacggcac ccccagcgga 600
atcgataacg cggttgcaac ctacggaaat gctttgctgt tcgagaagga ttcccacaat 660
ggaacgatta acacgaataa ctttaagttc ctcgacgatt tccccgcaat tcccatgatc 720
ctcacctaca cccgtatccc tcgttcgacc aaagacttgg tcgctcgcgt acgcgtgttg 780
gtcacggaaa agtttccaga ggtcatgaaa cctattcttg atgccatggg tgagtgcgcc 840
cttcagggcc tggagattat gactaagctg tccaagtgca agggtacgga cgacgaggca 900
gtcgaaacta acaatgaact ctacgagcag cttctggaat tgatccgcat caaccacggt 960
ctgctcgtgt ccatcggcgt gtctcaccca ggactggaac tgattaaaaa cctctcggat 1020
gacctccgta tcggctctac caagctgacc ggcgccggag gcggtggatg ctctctcacc 1080
ttgctccgac gcgatatcac ccaggagcag atcgattctt tcaaaaagaa actccaagat 1140
gacttctctt acgagacttt tgagactgac ctcggtggca ccggctgctg cttgctttcc 1200
gcgaagaact tgaataaaga tctcaaaatc aaatctctcg tcttccagct cttcgaaaac 1260
aagaccacca ccaagcaaca gatcgatgac ttgctccttc caggcaacac taaccttcct 1320
tggacaagc 1329
<210> 15
<211> 352
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 15
Met Ala Ser Glu Lys Glu Ile Arg Arg Glu Arg Phe Leu Asn Val Phe
1 5 10 15
Pro Lys Leu Val Glu Glu Leu Asn Ala Ser Leu Leu Ala Tyr Gly Met
20 25 30
Pro Lys Glu Ala Cys Asp Trp Tyr Ala His Ser Leu Asn Tyr Asn Thr
35 40 45
Pro Gly Gly Lys Leu Asn Arg Gly Leu Ser Val Val Asp Thr Tyr Ala
50 55 60
Ile Leu Ser Asn Lys Thr Val Glu Gln Leu Gly Gln Glu Glu Tyr Glu
65 70 75 80
Lys Val Ala Ile Leu Gly Trp Cys Ile Glu Leu Leu Gln Ala Tyr Trp
85 90 95
Leu Val Ala Asp Asp Met Met Asp Lys Ser Ile Thr Arg Arg Gly Gln
100 105 110
Pro Cys Trp Tyr Lys Val Pro Glu Val Gly Glu Ile Ala Ile Trp Asp
115 120 125
Ala Phe Met Leu Glu Ala Ala Ile Tyr Lys Leu Leu Lys Ser His Phe
130 135 140
Arg Asn Glu Lys Tyr Tyr Ile Asp Ile Thr Glu Leu Phe His Glu Val
145 150 155 160
Thr Phe Gln Thr Glu Leu Gly Gln Leu Met Asp Leu Ile Thr Ala Pro
165 170 175
Glu Asp Lys Val Asp Leu Ser Lys Phe Ser Leu Lys Lys His Ser Phe
180 185 190
Ile Val Thr Phe Lys Thr Ala Tyr Tyr Ser Phe Tyr Leu Pro Val Ala
195 200 205
Leu Ala Met Tyr Val Ala Gly Ile Thr Asp Glu Lys Asp Leu Lys Gln
210 215 220
Ala Arg Asp Val Leu Ile Pro Leu Gly Glu Tyr Phe Gln Ile Gln Asp
225 230 235 240
Asp Tyr Leu Asp Cys Phe Gly Thr Pro Glu Gln Ile Gly Lys Ile Gly
245 250 255
Thr Asp Ile Gln Asp Asn Lys Cys Ser Trp Val Ile Asn Lys Ala Leu
260 265 270
Glu Leu Ala Ser Ala Glu Gln Arg Lys Thr Leu Asp Glu Asn Tyr Gly
275 280 285
Lys Lys Asp Ser Val Ala Glu Ala Lys Cys Lys Lys Ile Phe Asn Asp
290 295 300
Leu Lys Ile Glu Gln Leu Tyr His Glu Tyr Glu Glu Ser Ile Ala Lys
305 310 315 320
Asp Leu Lys Ala Lys Ile Ser Gln Val Asp Glu Ser Arg Gly Phe Lys
325 330 335
Ala Asp Val Leu Thr Ala Phe Leu Asn Lys Val Tyr Lys Arg Ser Lys
340 345 350
<210> 16
<211> 1056
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 16
atggcatccg agaaggaaat ccgtcgtgaa cgattcctga acgtattccc aaagctcgtc 60
gaagagctta atgcttcctt gcttgcgtac ggaatgccga aggaagcgtg cgactggtat 120
gcgcattctc tcaactataa cacacctggc ggtaagctga atcgcggtct cagcgtggtg 180
gacacgtacg caatcctcag caataagacc gtggaacagc tgggacagga ggagtacgag 240
aaagttgcta tccttggatg gtgtatcgaa ctgctgcaag cctattggtt ggtagctgat 300
gacatgatgg ataagagcat cactcgacgt ggtcaaccat gctggtacaa ggtgccggag 360
gttggcgaaa tcgcgatctg ggatgcgttc atgttggagg ccgcaattta taagcttctc 420
aagtcccact ttcgtaacga aaagtactat attgacatca cagaactctt tcatgaagtg 480
accttccaaa ccgagctcgg ccaactcatg gatttgatca ccgcccccga agataaggtg 540
gacctttcga agttctccct caagaagcac tcgttcattg ttaccttcaa gacagcatac 600
tactcatttt acctgccagt cgccctcgca atgtacgtgg cgggcatcac tgatgaaaag 660
gatctgaagc aggcccgcga cgtccttatc cctcttggag agtacttcca gatccaggac 720
gattacttgg attgttttgg caccccagag cagattggta aaatcggtac tgatatccaa 780
gacaacaaat gttcctgggt aattaacaag gctctcgagt tggcttccgc ggaacaacgc 840
aagactcttg acgaaaatta cggtaagaaa gattcagtcg ctgaagccaa gtgcaagaaa 900
atctttaacg atcttaaaat cgaacagttg taccacgagt atgaagagtc tatcgctaaa 960
gacctgaaag ctaaaatttc ccaggtcgat gagtcacgcg gcttcaaagc ggatgtgctc 1020
accgccttcc ttaacaaggt gtataagcgc tccaaa 1056
<210> 17
<211> 491
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 17
Met Lys Leu Ser Thr Lys Leu Cys Trp Cys Gly Ile Lys Gly Arg Leu
1 5 10 15
Arg Pro Gln Lys Gln Gln Gln Leu His Asn Thr Asn Leu Gln Met Thr
20 25 30
Glu Leu Lys Lys Gln Lys Thr Ala Glu Gln Lys Thr Arg Pro Gln Asn
35 40 45
Val Gly Ile Lys Gly Ile Gln Ile Tyr Ile Pro Thr Gln Cys Val Asn
50 55 60
Gln Ser Glu Leu Glu Lys Phe Asp Gly Val Ser Gln Gly Lys Tyr Thr
65 70 75 80
Ile Gly Leu Gly Gln Thr Asn Met Ser Phe Val Asn Asp Arg Glu Asp
85 90 95
Ile Tyr Ser Met Ser Leu Thr Val Leu Ser Lys Leu Ile Lys Ser Tyr
100 105 110
Asn Ile Asp Thr Asn Lys Ile Gly Arg Leu Glu Val Gly Thr Glu Thr
115 120 125
Leu Ile Asp Lys Ser Lys Ser Val Lys Ser Val Leu Met Gln Leu Phe
130 135 140
Gly Glu Asn Thr Asp Val Glu Gly Ile Asp Thr Leu Asn Ala Cys Tyr
145 150 155 160
Gly Gly Thr Asn Ala Leu Phe Asn Ser Leu Asn Trp Ile Glu Ser Asn
165 170 175
Ala Trp Asp Gly Arg Asp Ala Ile Val Val Cys Gly Asp Ile Ala Ile
180 185 190
Tyr Asp Lys Gly Ala Ala Arg Pro Thr Gly Gly Ala Gly Thr Val Ala
195 200 205
Met Trp Ile Gly Pro Asp Ala Pro Ile Val Phe Asp Ser Val Arg Ala
210 215 220
Ser Tyr Met Glu His Ala Tyr Asp Phe Tyr Lys Pro Asp Phe Thr Ser
225 230 235 240
Glu Tyr Pro Tyr Val Asp Gly His Phe Ser Leu Thr Cys Tyr Val Lys
245 250 255
Ala Leu Asp Gln Val Tyr Lys Ser Tyr Ser Lys Lys Ala Ile Ser Lys
260 265 270
Gly Leu Val Ser Asp Pro Ala Gly Ser Asp Ala Leu Asn Val Leu Lys
275 280 285
Tyr Phe Asp Tyr Asn Val Phe His Val Pro Thr Cys Lys Leu Val Thr
290 295 300
Lys Ser Tyr Gly Arg Leu Leu Tyr Asn Asp Phe Arg Ala Asn Pro Gln
305 310 315 320
Leu Phe Pro Glu Val Asp Ala Glu Leu Ala Thr Arg Asp Tyr Asp Glu
325 330 335
Ser Leu Thr Asp Lys Asn Ile Glu Lys Thr Phe Val Asn Val Ala Lys
340 345 350
Pro Phe His Lys Glu Arg Val Ala Gln Ser Leu Ile Val Pro Thr Asn
355 360 365
Thr Gly Asn Met Tyr Thr Ala Ser Val Tyr Ala Ala Phe Ala Ser Leu
370 375 380
Leu Asn Tyr Val Gly Ser Asp Asp Leu Gln Gly Lys Arg Val Gly Leu
385 390 395 400
Phe Ser Tyr Gly Ser Gly Leu Ala Ala Ser Leu Tyr Ser Cys Lys Ile
405 410 415
Val Gly Asp Val Gln His Ile Ile Lys Glu Leu Asp Ile Thr Asn Lys
420 425 430
Leu Ala Lys Arg Ile Thr Glu Thr Pro Lys Asp Tyr Glu Ala Ala Ile
435 440 445
Glu Leu Arg Glu Asn Ala His Leu Lys Lys Asn Phe Lys Pro Gln Gly
450 455 460
Ser Ile Glu His Leu Gln Ser Gly Val Tyr Tyr Leu Thr Asn Ile Asp
465 470 475 480
Asp Lys Phe Arg Arg Ser Tyr Asp Val Lys Lys
485 490
<210> 18
<211> 1473
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 18
atgaaattgt ctaccaagct ctgttggtgt ggcatcaaag gccgtcttcg cccacagaaa 60
cagcaacagc tgcacaatac caacctccag atgaccgagc ttaaaaagca aaagacggca 120
gagcagaaga ctcgccctca gaatgtcggc attaagggca ttcaaatcta catcccaacc 180
cagtgcgtaa atcagagcga gcttgaaaag ttcgatggag tatcccaggg aaagtataca 240
attggactgg gccagaccaa catgtcgttc gtgaacgacc gcgaagacat ttattcgatg 300
tcgcttactg tgttgagcaa attgatcaaa tcctacaaca ttgatactaa taaaattggt 360
cgcctcgagg tcggcaccga aaccctgatc gataagtcga agagcgtgaa aagcgtgctg 420
atgcaacttt tcggtgagaa caccgatgtg gaaggaatcg atacccttaa cgcatgttac 480
ggtggaacca acgccttgtt taattcgctt aactggatcg aatcgaacgc atgggatggc 540
cgcgatgcaa tcgtggtttg cggagatatc gccatttacg acaagggcgc cgcgcgcccc 600
acgggcggcg cgggcacagt ggcaatgtgg atcggtcctg atgcaccaat tgtgttcgat 660
tcggtccgag caagctatat ggagcatgcc tatgatttct acaaaccaga ttttaccagc 720
gaatacccct acgtcgatgg tcacttctcc ttgacatgct acgtcaaggc gcttgaccag 780
gtgtacaaga gctactccaa aaaggcgatc tccaaaggcc tggtgtccga tcctgcaggt 840
tcagatgctc ttaatgttct taagtatttc gattacaacg ttttccacgt gcccacgtgc 900
aaactcgtga cgaagtcata tggtcgcctc ttgtataacg attttcgagc taatccacag 960
ctttttccgg aagtggacgc agagctggcc acccgcgact acgacgagtc tctcacggat 1020
aagaatatcg aaaaaacttt cgtgaatgtg gctaaaccat tccataagga acgcgtcgca 1080
caatctctca ttgttcccac gaacaccgga aacatgtaca cggcctcagt gtatgctgca 1140
ttcgcgtcac tgcttaacta tgttggatct gatgacctgc aaggtaaacg cgttggactc 1200
ttctcttacg gttcgggttt ggcagcttcg ctgtactcct gcaagattgt cggagacgtc 1260
cagcatatca ttaaagaact ggacattaca aacaagctcg cgaaacgcat cactgaaacg 1320
cccaaggatt acgaagcggc cattgaattg cgcgagaacg cacatttgaa gaaaaacttt 1380
aaaccgcaag gatccatcga acacctgcaa tcgggtgtct attatctcac caacattgac 1440
gataagtttc gacgttccta tgatgttaaa aag 1473
<210> 19
<211> 398
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 19
Met Ser Gln Asn Val Tyr Ile Val Ser Thr Ala Arg Thr Pro Ile Gly
1 5 10 15
Ser Phe Gln Gly Ser Leu Ser Ser Lys Thr Ala Val Glu Leu Gly Ala
20 25 30
Val Ala Leu Lys Gly Ala Leu Ala Lys Val Pro Glu Leu Asp Ala Ser
35 40 45
Lys Asp Phe Asp Glu Ile Ile Phe Gly Asn Val Leu Ser Ala Asn Leu
50 55 60
Gly Gln Ala Pro Ala Arg Gln Val Ala Leu Ala Ala Gly Leu Ser Asn
65 70 75 80
His Ile Val Ala Ser Thr Val Asn Lys Val Cys Ala Ser Ala Met Lys
85 90 95
Ala Ile Ile Leu Gly Ala Gln Ser Ile Lys Cys Gly Asn Ala Asp Val
100 105 110
Val Val Ala Gly Gly Cys Glu Ser Met Thr Asn Ala Pro Tyr Tyr Met
115 120 125
Pro Ala Ala Arg Ala Gly Ala Lys Phe Gly Gln Thr Val Leu Val Asp
130 135 140
Gly Val Glu Arg Asp Gly Leu Asn Asp Ala Tyr Asp Gly Leu Ala Met
145 150 155 160
Gly Val His Ala Glu Lys Cys Ala Arg Asp Trp Asp Ile Thr Arg Glu
165 170 175
Gln Gln Asp Asn Phe Ala Ile Glu Ser Tyr Gln Lys Ser Gln Lys Ser
180 185 190
Gln Lys Glu Gly Lys Phe Asp Asn Glu Ile Val Pro Val Thr Ile Lys
195 200 205
Gly Phe Arg Gly Lys Pro Asp Thr Gln Val Thr Lys Asp Glu Glu Pro
210 215 220
Ala Arg Leu His Val Glu Lys Leu Arg Ser Ala Arg Thr Val Phe Gln
225 230 235 240
Lys Glu Asn Gly Thr Val Thr Ala Ala Asn Ala Ser Pro Ile Asn Asp
245 250 255
Gly Ala Ala Ala Val Ile Leu Val Ser Glu Lys Val Leu Lys Glu Lys
260 265 270
Asn Leu Lys Pro Leu Ala Ile Ile Lys Gly Trp Gly Glu Ala Ala His
275 280 285
Gln Pro Ala Asp Phe Thr Trp Ala Pro Ser Leu Ala Val Pro Lys Ala
290 295 300
Leu Lys His Ala Gly Ile Glu Asp Ile Asn Ser Val Asp Tyr Phe Glu
305 310 315 320
Phe Asn Glu Ala Phe Ser Val Val Gly Leu Val Asn Thr Lys Ile Leu
325 330 335
Lys Leu Asp Pro Ser Lys Val Asn Val Tyr Gly Gly Ala Val Ala Leu
340 345 350
Gly His Pro Leu Gly Cys Ser Gly Ala Arg Val Val Val Thr Leu Leu
355 360 365
Ser Ile Leu Gln Gln Glu Gly Gly Lys Ile Gly Val Ala Ala Ile Cys
370 375 380
Asn Gly Gly Gly Gly Ala Ser Ser Ile Val Ile Glu Lys Ile
385 390 395
<210> 20
<211> 1194
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 20
atgtcacaga atgtctacat cgtctctacc gcccgcaccc cgattggttc cttccagggt 60
tctcttagct cgaaaacagc ggttgaactc ggcgccgttg cactgaaagg agccttggct 120
aaagtaccag aattggacgc gtccaaggac ttcgatgaaa ttattttcgg taacgtactg 180
tcagctaatc tcggccaagc tccagctcgc caggtcgcgc tcgctgcggg cctctccaat 240
cacattgtgg ccagcactgt gaataaggtc tgcgcatcgg cgatgaaggc gattatcctg 300
ggcgcccaga gcattaaatg cggtaatgca gatgttgtgg ttgccggtgg ctgtgagtcg 360
atgaccaacg ctccgtacta catgcccgct gcacgcgctg gcgctaaatt cggtcaaacc 420
gtcctggtcg atggtgtcga gcgcgatggc ctcaacgatg cgtacgacgg cctcgcgatg 480
ggagtacacg ctgagaagtg tgcccgcgac tgggatatca cccgagagca gcaggacaat 540
tttgccatcg agagctatca gaagtcacaa aagtcacaga aggagggtaa attcgataac 600
gaaatcgttc cagtgacgat caaaggcttc cgcggtaagc cagatactca agtaaccaaa 660
gatgaggaac ctgcccgctt gcacgtagag aagctccgct cggcccgcac tgtcttccag 720
aaagaaaatg gaaccgtgac ggcagctaac gcaagcccta ttaacgacgg tgcggcagcg 780
gtcattctcg tgtcagaaaa ggtgctgaaa gaaaagaacc tcaaacctct ggccatcatc 840
aagggctggg gtgaggccgc tcaccaacct gctgacttca cctgggcacc atccctggct 900
gtaccaaaag cactcaagca cgctggtatt gaagatatca actccgtcga ctacttcgaa 960
tttaatgagg cattctcggt tgtcggcctt gttaacacta agattcttaa gctcgatcca 1020
tccaaggtca atgtatatgg cggtgcggtt gctttgggcc atcctctcgg ttgctcagga 1080
gcccgcgtgg ttgttactct cctcagcatt cttcagcagg agggtggaaa gatcggcgtc 1140
gctgctattt gcaacggcgg tggtggtgcc tcctcaattg ttattgaaaa gatt 1194
<210> 21
<211> 1045
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 21
Met Ser Leu Pro Leu Lys Thr Ile Val His Leu Val Lys Pro Phe Ala
1 5 10 15
Cys Thr Ala Arg Phe Ser Ala Arg Tyr Pro Ile His Val Ile Val Val
20 25 30
Ala Val Leu Leu Ser Ala Ala Ala Tyr Leu Ser Val Thr Gln Ser Tyr
35 40 45
Leu Asn Glu Trp Lys Leu Asp Ser Asn Gln Tyr Ser Thr Tyr Leu Ser
50 55 60
Ile Lys Pro Asp Glu Leu Phe Glu Lys Cys Thr His Tyr Tyr Arg Ser
65 70 75 80
Pro Val Ser Asp Thr Trp Lys Leu Leu Ser Ser Lys Glu Ala Ala Asp
85 90 95
Ile Tyr Thr Pro Phe His Tyr Tyr Leu Ser Thr Ile Ser Phe Gln Ser
100 105 110
Lys Asp Asn Ser Thr Thr Leu Pro Ser Leu Asp Asp Val Ile Tyr Ser
115 120 125
Val Asp His Thr Arg Tyr Leu Leu Ser Glu Glu Pro Lys Ile Pro Thr
130 135 140
Glu Leu Val Ser Glu Asn Gly Thr Lys Trp Arg Leu Arg Asn Asn Ser
145 150 155 160
Asn Phe Ile Leu Asp Leu His Asn Ile Tyr Arg Asn Met Val Lys Gln
165 170 175
Phe Ser Asn Lys Thr Ser Glu Phe Asp Gln Phe Asp Leu Phe Ile Ile
180 185 190
Leu Ala Ala Tyr Leu Thr Leu Phe Tyr Thr Leu Cys Cys Leu Phe Asn
195 200 205
Asp Met Arg Lys Ile Gly Ser Lys Phe Trp Leu Ser Phe Ser Ala Leu
210 215 220
Ser Asn Ser Ala Cys Ala Leu Tyr Leu Ser Leu Tyr Thr Thr His Ser
225 230 235 240
Leu Leu Lys Lys Pro Ala Ser Leu Leu Ser Leu Val Ile Gly Leu Pro
245 250 255
Phe Ile Val Val Ile Ile Gly Phe Lys His Lys Val Arg Leu Ala Ala
260 265 270
Phe Ser Leu Gln Lys Phe His Arg Ile Ser Ile Asp Lys Lys Ile Thr
275 280 285
Val Ser Asn Ile Ile Tyr Glu Ala Met Phe Gln Glu Gly Ala Tyr Leu
290 295 300
Ile Arg Asp Tyr Leu Phe Tyr Ile Ser Ser Phe Ile Gly Cys Ala Ile
305 310 315 320
Tyr Ala Arg His Leu Pro Gly Leu Val Asn Phe Cys Ile Leu Ser Thr
325 330 335
Phe Met Leu Val Phe Asp Leu Leu Leu Ser Ala Thr Phe Tyr Ser Ala
340 345 350
Ile Leu Ser Met Lys Leu Glu Ile Asn Ile Ile His Arg Ser Thr Val
355 360 365
Ile Arg Gln Thr Leu Glu Glu Asp Gly Val Val Pro Thr Thr Ala Asp
370 375 380
Ile Ile Tyr Lys Asp Glu Thr Ala Ser Glu Pro His Phe Leu Arg Ser
385 390 395 400
Asn Val Ala Ile Ile Leu Gly Lys Ala Ser Val Ile Gly Leu Leu Leu
405 410 415
Leu Ile Asn Leu Tyr Val Phe Thr Asp Lys Leu Asn Ala Thr Ile Leu
420 425 430
Asn Thr Val Tyr Phe Asp Ser Thr Ile Tyr Ser Leu Pro Asn Phe Ile
435 440 445
Asn Tyr Lys Asp Ile Gly Asn Leu Ser Asn Gln Val Ile Ile Ser Val
450 455 460
Leu Pro Lys Gln Tyr Tyr Thr Pro Leu Lys Lys Tyr His Gln Ile Glu
465 470 475 480
Asp Ser Val Leu Leu Ile Ile Asp Ser Val Ser Asn Ala Ile Arg Asp
485 490 495
Gln Phe Ile Ser Lys Leu Leu Phe Phe Ala Phe Ala Val Ser Ile Ser
500 505 510
Ile Asn Val Tyr Leu Leu Asn Ala Ala Lys Ile His Thr Gly Tyr Met
515 520 525
Asn Phe Gln Pro Gln Ser Asn Lys Ile Asp Asp Leu Val Val Gln Gln
530 535 540
Lys Ser Ala Thr Ile Glu Phe Ser Glu Thr Arg Ser Met Pro Ala Ser
545 550 555 560
Ser Gly Leu Glu Thr Pro Val Thr Ala Lys Asp Ile Ile Ile Ser Glu
565 570 575
Glu Ile Gln Asn Asn Glu Cys Val Tyr Ala Leu Ser Ser Gln Asp Glu
580 585 590
Pro Ile Arg Pro Leu Ser Asn Leu Val Glu Leu Met Glu Lys Glu Gln
595 600 605
Leu Lys Asn Met Asn Asn Thr Glu Val Ser Asn Leu Val Val Asn Gly
610 615 620
Lys Leu Pro Leu Tyr Ser Leu Glu Lys Lys Leu Glu Asp Thr Thr Arg
625 630 635 640
Ala Val Leu Val Arg Arg Lys Ala Leu Ser Thr Leu Ala Glu Ser Pro
645 650 655
Ile Leu Val Ser Glu Lys Leu Pro Phe Arg Asn Tyr Asp Tyr Asp Arg
660 665 670
Val Phe Gly Ala Cys Cys Glu Asn Val Ile Gly Tyr Met Pro Ile Pro
675 680 685
Val Gly Val Ile Gly Pro Leu Ile Ile Asp Gly Thr Ser Tyr His Ile
690 695 700
Pro Met Ala Thr Thr Glu Gly Cys Leu Val Ala Ser Ala Met Arg Gly
705 710 715 720
Cys Lys Ala Ile Asn Ala Gly Gly Gly Ala Thr Thr Val Leu Thr Lys
725 730 735
Asp Gly Met Thr Arg Gly Pro Val Val Arg Phe Pro Thr Leu Ile Arg
740 745 750
Ser Gly Ala Cys Lys Ile Trp Leu Asp Ser Glu Glu Gly Gln Asn Ser
755 760 765
Ile Lys Lys Ala Phe Asn Ser Thr Ser Arg Phe Ala Arg Leu Gln His
770 775 780
Ile Gln Thr Cys Leu Ala Gly Asp Leu Leu Phe Met Arg Phe Arg Thr
785 790 795 800
Thr Thr Gly Asp Ala Met Gly Met Asn Met Ile Ser Lys Gly Val Glu
805 810 815
Tyr Ser Leu Lys Gln Met Val Glu Glu Tyr Gly Trp Glu Asp Met Glu
820 825 830
Val Val Ser Val Ser Gly Asn Tyr Cys Thr Asp Lys Lys Pro Ala Ala
835 840 845
Ile Asn Trp Ile Glu Gly Arg Gly Lys Ser Val Val Ala Glu Ala Thr
850 855 860
Ile Pro Gly Asp Val Val Lys Ser Val Leu Lys Ser Asp Val Ser Ala
865 870 875 880
Leu Val Glu Leu Asn Ile Ser Lys Asn Leu Val Gly Ser Ala Met Ala
885 890 895
Gly Ser Val Gly Gly Phe Asn Ala His Ala Ala Asn Leu Val Thr Ala
900 905 910
Leu Phe Leu Ala Leu Gly Gln Asp Pro Ala Gln Asn Val Glu Ser Ser
915 920 925
Asn Cys Ile Thr Leu Met Lys Glu Val Asp Gly Asp Leu Arg Ile Ser
930 935 940
Val Ser Met Pro Ser Ile Glu Val Gly Thr Ile Gly Gly Gly Thr Val
945 950 955 960
Leu Glu Pro Gln Gly Ala Met Leu Asp Leu Leu Gly Val Arg Gly Pro
965 970 975
His Pro Thr Glu Pro Gly Ala Asn Ala Arg Gln Leu Ala Arg Ile Ile
980 985 990
Ala Cys Ala Val Leu Ala Gly Glu Leu Ser Leu Cys Ser Ala Leu Ala
995 1000 1005
Ala Gly His Leu Val Gln Ser His Met Thr His Asn Arg Lys Thr
1010 1015 1020
Asn Lys Ala Asn Glu Leu Pro Gln Pro Ser Asn Lys Gly Pro Pro
1025 1030 1035
Cys Lys Thr Ser Ala Leu Leu
1040 1045
<210> 22
<211> 3135
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 22
atgtccttgc ctcttaagac catcgtgcac ctggtaaagc ctttcgcctg cactgcccgc 60
ttctccgcgc gctaccccat ccacgttatc gtggtggcgg tgctgctctc ggcggcagct 120
tacttgagcg taacccaatc ctacttgaat gaatggaagc tcgactcgaa ccagtactcc 180
acctacctta gcatcaagcc tgatgaattg ttcgaaaaat gtacccacta ctaccgctct 240
ccagtgtctg atacttggaa actccttagc tctaaggaag ctgcagatat ttacaccccc 300
ttccattatt acctgtccac catttccttc cagtccaagg acaactcgac caccctgccc 360
agcctggatg acgttatcta ctccgtagac catacccgct acttgctcag cgaggaaccg 420
aaaatcccga cggaactcgt atccgaaaac ggaaccaagt ggcgtctgcg taataactct 480
aactttatcc ttgatctcca caacatttat cgcaatatgg ttaagcagtt ctcaaacaaa 540
acctcggaat ttgaccaatt cgacttgttc atcattctcg cagcatattt gactttgttt 600
tacaccctct gctgtctctt taatgacatg cgcaaaatcg gctccaagtt ctggctttcc 660
ttctcagctc tttccaacag cgcctgcgca ctttatttga gcttgtacac tacccactcc 720
ctccttaaga aaccagcttc cctcctctcc ctcgtcatcg gcttgccatt catcgtggtt 780
atcattggtt tcaaacataa ggtgcgcctc gctgcgttct ccctgcagaa gtttcaccga 840
attagcatcg ataaaaagat cacggtctct aacattatct acgaagccat gttccaagaa 900
ggcgcgtacc ttatccgaga ttacctgttc tacatttcca gcttcatcgg ttgtgctatc 960
tacgcacgcc accttccagg tctggtgaat ttctgcattc tctccacctt tatgcttgtg 1020
tttgatctcc ttttgagcgc cactttctat agcgcgatcc tctcaatgaa actggaaatc 1080
aacatcattc accgcagcac cgtcattcgc cagacccttg aagaagatgg cgtggtgccg 1140
accaccgccg atattattta taaagatgaa accgcatccg aaccacattt cctgcgttcc 1200
aacgtggcga ttatccttgg taaggcgtcg gtcattggac tgctgctgct gattaatttg 1260
tacgttttca cagataagct caacgcgacg attctgaata ctgtttactt cgattctacc 1320
atctattccc tcccgaactt catcaactat aaggatatcg gaaacctttc taatcaagtc 1380
attattagcg ttctgccaaa gcagtactac acccctttga aaaagtatca tcagattgag 1440
gactctgtgc tccttatcat cgactccgtg agcaatgcta ttcgcgatca atttatctcc 1500
aagctcttgt ttttcgcgtt cgctgtttca atttccatca acgtgtacct gttgaacgca 1560
gctaagattc acacgggcta catgaatttc cagccccaat ccaacaagat tgatgacctt 1620
gtcgtgcaac agaagtctgc aaccattgaa ttctccgaga cccgatccat gcctgcatcc 1680
tccggcctcg agaccccagt taccgcgaag gacatcatta tttcagaaga aatccagaat 1740
aatgaatgcg tatacgcgct gtcatctcag gatgaaccca ttcgcccgtt gtcgaacctt 1800
gttgaactga tggagaaaga gcaactgaaa aatatgaaca acacagaagt ttctaacttg 1860
gtggtgaacg gtaaattgcc gttgtactcc ctggaaaaga aactcgaaga taccacccga 1920
gccgttcttg ttcgacgcaa ggcgctttcc accctggccg aatctccaat ccttgtttcc 1980
gaaaaactcc cttttcgcaa ctacgattac gatcgtgtgt tcggcgcgtg ttgcgagaac 2040
gtaattggct acatgcctat cccagtagga gtgatcggcc ctctcatcat cgatggcacc 2100
agctaccata ttccaatggc aaccacagaa ggttgcttgg tagcaagcgc gatgcgagga 2160
tgcaaagcaa tcaacgctgg aggcggtgct accaccgttc tgaccaagga tggaatgact 2220
cgcggccccg tagtccgctt ccctactctc atccgctctg gtgcgtgcaa gatttggctc 2280
gattccgagg aaggccagaa ctctatcaaa aaagcgttca acagcacctc ccgcttcgcc 2340
cgtttgcaac acattcagac ttgcctggca ggcgacctcc tcttcatgcg tttccgcacc 2400
actactggcg acgcaatggg tatgaacatg atctccaaag gagttgagta ctcgctcaaa 2460
cagatggtgg aagagtatgg ctgggaagac atggaagttg tgtccgtttc cggtaactac 2520
tgcacggaca agaaaccagc tgcgatcaat tggattgagg gacgcggcaa atctgtcgtc 2580
gcggaggcaa ccatcccagg tgacgtcgtc aaatccgttc tcaaatctga cgtctccgca 2640
ctggttgaac tgaacatctc taagaacctt gtgggttctg ctatggctgg cagcgtcggt 2700
ggctttaacg cgcatgcggc gaaccttgtt accgcgctct tcctggcgct cggacaggat 2760
cccgcgcaga atgtggagtc atcaaattgc attacactga tgaaggaggt ggacggcgat 2820
ctgcgtatct ctgtttccat gccgtccatt gaggtgggta ccattggtgg cggcacagtc 2880
ttggagcctc agggtgccat gctggatctg ttgggcgtac gtggccccca tcctaccgaa 2940
ccaggcgcaa acgcacgcca gctcgcgcgc attatcgcat gcgccgtgct ggcaggtgaa 3000
ttgtccctct gctcagcctt ggccgcgggt catttggtcc aaagccacat gacccataac 3060
cgcaaaacga ataaggccaa cgaacttcct caaccttcaa acaagggtcc accttgcaaa 3120
acctccgcgc tgttg 3135
<210> 23
<211> 451
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 23
Met Ser Glu Leu Arg Ala Phe Ser Ala Pro Gly Lys Ala Leu Leu Ala
1 5 10 15
Gly Gly Tyr Leu Val Leu Asp Thr Lys Tyr Glu Ala Phe Val Val Gly
20 25 30
Leu Ser Ala Arg Met His Ala Val Ala His Pro Tyr Gly Ser Leu Gln
35 40 45
Gly Ser Asp Lys Phe Glu Val Arg Val Lys Ser Lys Gln Phe Lys Asp
50 55 60
Gly Glu Trp Leu Tyr His Ile Ser Pro Lys Ser Gly Phe Ile Pro Val
65 70 75 80
Ser Ile Gly Gly Ser Lys Asn Pro Phe Ile Glu Lys Val Ile Ala Asn
85 90 95
Val Phe Ser Tyr Phe Lys Pro Asn Met Asp Asp Tyr Cys Asn Arg Asn
100 105 110
Leu Phe Val Ile Asp Ile Phe Ser Asp Asp Ala Tyr His Ser Gln Glu
115 120 125
Asp Ser Val Thr Glu His Arg Gly Asn Arg Arg Leu Ser Phe His Ser
130 135 140
His Arg Ile Glu Glu Val Pro Lys Thr Gly Leu Gly Ser Ser Ala Gly
145 150 155 160
Leu Val Thr Val Leu Thr Thr Ala Leu Ala Ser Phe Phe Val Ser Asp
165 170 175
Leu Glu Asn Asn Val Asp Lys Tyr Arg Glu Val Ile His Asn Leu Ala
180 185 190
Gln Val Ala His Cys Gln Ala Gln Gly Lys Ile Gly Ser Gly Phe Asp
195 200 205
Val Ala Ala Ala Ala Tyr Gly Ser Ile Arg Tyr Arg Arg Phe Pro Pro
210 215 220
Ala Leu Ile Ser Asn Leu Pro Asp Ile Gly Ser Ala Thr Tyr Gly Ser
225 230 235 240
Lys Leu Ala His Leu Val Asp Glu Glu Asp Trp Asn Ile Thr Ile Lys
245 250 255
Ser Asn His Leu Pro Ser Gly Leu Thr Leu Trp Met Gly Asp Ile Lys
260 265 270
Asn Gly Ser Glu Thr Val Lys Leu Val Gln Lys Val Lys Asn Trp Tyr
275 280 285
Asp Ser His Met Pro Glu Ser Leu Lys Ile Tyr Thr Glu Leu Asp His
290 295 300
Ala Asn Ser Arg Phe Met Asp Gly Leu Ser Lys Leu Asp Arg Leu His
305 310 315 320
Glu Thr His Asp Asp Tyr Ser Asp Gln Ile Phe Glu Ser Leu Glu Arg
325 330 335
Asn Asp Cys Thr Cys Gln Lys Tyr Pro Glu Ile Thr Glu Val Arg Asp
340 345 350
Ala Val Ala Thr Ile Arg Arg Ser Phe Arg Lys Ile Thr Lys Glu Ser
355 360 365
Gly Ala Asp Ile Glu Pro Pro Val Gln Thr Ser Leu Leu Asp Asp Cys
370 375 380
Gln Thr Leu Lys Gly Val Leu Thr Cys Leu Ile Pro Gly Ala Gly Gly
385 390 395 400
Tyr Asp Ala Ile Ala Val Ile Thr Lys Gln Asp Val Asp Leu Arg Ala
405 410 415
Gln Thr Ala Asn Asp Lys Arg Phe Ser Lys Val Gln Trp Leu Asp Val
420 425 430
Thr Gln Ala Asp Trp Gly Val Arg Lys Glu Lys Asp Pro Glu Thr Tyr
435 440 445
Leu Asp Lys
450
<210> 24
<211> 1353
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 24
atgtccgaac tgcgcgcatt ttccgcgcct ggtaaggcgc tgctcgccgg cggctacctt 60
gtgttggaca ctaaatacga ggccttcgtg gtgggccttt ccgcacgtat gcacgccgtc 120
gctcatcctt atggttcgtt gcaaggttcc gataagtttg aggtgcgcgt gaaatccaaa 180
cagtttaaag atggtgaatg gctgtaccac atcagcccaa agtcaggctt cattccagta 240
tccattggtg gttccaagaa tccatttatc gagaaggtca ttgccaacgt cttttcatac 300
ttcaaaccaa acatggatga ttattgtaat cgcaacttgt tcgtgatcga tatcttctct 360
gacgacgctt atcactcgca ggaagattcc gttacggagc atcgtggaaa ccgacgtctg 420
tcatttcatt cgcaccgcat cgaggaagtt cccaagaccg gcctgggctc gtctgccggc 480
cttgtcactg ttttgactac cgccctcgca tctttcttcg tgtctgacct tgaaaacaac 540
gttgataagt accgcgaggt gattcacaac ctggcgcagg tagcgcactg ccaggcgcag 600
ggaaaaattg gcagcggctt tgatgtggct gctgctgcat acggcagcat ccgttaccgc 660
cgcttcccac cagcgcttat tagcaacctg ccggacatcg gctccgcgac ttacggttcc 720
aaattggccc accttgtcga cgaggaagac tggaacatca caatcaagtc taaccacctc 780
ccttccggcc tgacgctctg gatgggcgat atcaagaatg gctctgaaac cgtgaagctc 840
gtgcagaagg ttaagaactg gtacgactca cacatgcctg aatcgttgaa aatctacacc 900
gagcttgatc acgcgaactc ccgtttcatg gacggattgt cgaagttgga ccgtctgcat 960
gaaacgcacg acgattattc cgaccaaatc tttgaatcgc tggaacgtaa cgattgtact 1020
tgtcagaagt atcccgaaat caccgaggtc cgcgacgcgg tcgcgacaat tcgccgctca 1080
ttccgcaaaa tcactaagga atccggtgct gacattgagc ccccggttca gacctccctg 1140
ctcgatgatt gccagactct caagggtgtc ttgacgtgtt tgatccccgg agccggcggc 1200
tacgatgcca tcgcagttat caccaagcag gatgtggatc tccgcgcgca gacagcaaac 1260
gataagcgct tttcaaaggt gcagtggctg gacgtgaccc aagctgattg gggcgttcgt 1320
aaagaaaaag atccagagac atatctggat aag 1353
<210> 25
<211> 288
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 25
Met Thr Ala Asp Asn Asn Ser Met Pro His Gly Ala Val Ser Ser Tyr
1 5 10 15
Ala Lys Leu Val Gln Asn Gln Thr Pro Glu Asp Ile Leu Glu Glu Phe
20 25 30
Pro Glu Ile Ile Pro Leu Gln Gln Arg Pro Asn Thr Arg Ser Ser Glu
35 40 45
Thr Ser Asn Asp Glu Ser Gly Glu Thr Cys Phe Ser Gly His Asp Glu
50 55 60
Glu Gln Ile Lys Leu Met Asn Glu Asn Cys Ile Val Leu Asp Trp Asp
65 70 75 80
Asp Asn Ala Ile Gly Ala Gly Thr Lys Lys Val Cys His Leu Met Glu
85 90 95
Asn Ile Glu Lys Gly Leu Leu His Arg Ala Phe Ser Val Phe Ile Phe
100 105 110
Asn Glu Gln Gly Glu Leu Leu Leu Gln Gln Arg Ala Thr Glu Lys Ile
115 120 125
Thr Phe Pro Asp Leu Trp Thr Asn Thr Cys Cys Ser His Pro Leu Cys
130 135 140
Ile Asp Asp Glu Leu Gly Leu Lys Gly Lys Leu Asp Asp Lys Ile Lys
145 150 155 160
Gly Ala Ile Thr Ala Ala Val Arg Lys Leu Asp His Glu Leu Gly Ile
165 170 175
Pro Glu Asp Glu Thr Lys Thr Arg Gly Lys Phe His Phe Leu Asn Arg
180 185 190
Ile His Tyr Met Ala Pro Ser Asn Glu Pro Trp Gly Glu His Glu Ile
195 200 205
Asp Tyr Ile Leu Phe Tyr Lys Ile Asn Ala Lys Glu Asn Leu Thr Val
210 215 220
Asn Pro Asn Val Asn Glu Val Arg Asp Phe Lys Trp Val Ser Pro Asn
225 230 235 240
Asp Leu Lys Thr Met Phe Ala Asp Pro Ser Tyr Lys Phe Thr Pro Trp
245 250 255
Phe Lys Ile Ile Cys Glu Asn Tyr Leu Phe Asn Trp Trp Glu Gln Leu
260 265 270
Asp Asp Leu Ser Glu Val Glu Asn Asp Arg Gln Ile His Arg Met Leu
275 280 285
<210> 26
<211> 864
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 26
atgacagcag ataacaattc catgcctcac ggtgctgtgt cttcatacgc gaaactggtg 60
cagaaccaaa cacctgagga tattctcgag gaattccccg aaatcatccc tctccagcag 120
cgcccgaaca cccgctcctc tgaaacctct aatgacgaat ccggcgagac atgtttctcc 180
ggccacgacg aagagcaaat taagctgatg aacgaaaatt gcatcgtgtt ggactgggat 240
gacaacgcca ttggagccgg taccaaaaag gtgtgccatc tgatggagaa cattgaaaag 300
ggactgctcc atcgagcttt ctcagtgttc attttcaatg aacagggtga gctcctgctg 360
caacagcgcg caacggaaaa gattaccttc cccgacctct ggactaacac ctgctgctca 420
cacccattgt gtatcgatga cgaacttggc ttgaagggca agcttgatga taagattaag 480
ggcgcaatca cggccgctgt tcgtaaactc gatcacgaac ttggaattcc tgaagacgaa 540
accaagactc gcggcaaatt tcacttcttg aatcgtatcc actatatggc gccttccaac 600
gagccttggg gcgagcacga gatcgactat attttgttct acaagatcaa tgcaaaggag 660
aacctgactg tgaacccgaa tgtgaatgaa gtgcgtgact tcaagtgggt ctcaccgaac 720
gatctgaaga ccatgtttgc cgatccatcc tacaaattca ccccgtggtt caaaattatc 780
tgtgaaaact atctgttcaa ttggtgggaa caactcgacg atttgtccga ggtcgagaac 840
gatcgtcaaa tccaccgtat gctt 864
<210> 27
<211> 527
<212> PRT
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 27
Met Ala Ala Asp Gln Leu Val Lys Thr Glu Val Thr Lys Lys Ser Phe
1 5 10 15
Thr Ala Pro Val Gln Lys Ala Ser Thr Pro Val Leu Thr Asn Lys Thr
20 25 30
Val Ile Ser Gly Ser Lys Val Lys Ser Leu Ser Ser Ala Gln Ser Ser
35 40 45
Ser Ser Gly Pro Ser Ser Ser Ser Glu Glu Asp Asp Ser Arg Asp Ile
50 55 60
Glu Ser Leu Asp Lys Lys Ile Arg Pro Leu Glu Glu Leu Glu Ala Leu
65 70 75 80
Leu Ser Ser Gly Asn Thr Lys Gln Leu Lys Asn Lys Glu Val Ala Ala
85 90 95
Leu Val Ile His Gly Lys Leu Pro Leu Tyr Ala Leu Glu Lys Lys Leu
100 105 110
Gly Asp Thr Thr Arg Ala Val Ala Val Arg Arg Lys Ala Leu Ser Ile
115 120 125
Leu Ala Glu Ala Pro Val Leu Ala Ser Asp Arg Leu Pro Tyr Lys Asn
130 135 140
Tyr Asp Tyr Asp Arg Val Phe Gly Ala Cys Cys Glu Asn Val Ile Gly
145 150 155 160
Tyr Met Pro Leu Pro Val Gly Val Ile Gly Pro Leu Val Ile Asp Gly
165 170 175
Thr Ser Tyr His Ile Pro Met Ala Thr Thr Glu Gly Cys Leu Val Ala
180 185 190
Ser Ala Met Arg Gly Cys Lys Ala Ile Asn Ala Gly Gly Gly Ala Thr
195 200 205
Thr Val Leu Thr Lys Asp Gly Met Thr Arg Gly Pro Val Val Arg Phe
210 215 220
Pro Thr Leu Lys Arg Ser Gly Ala Cys Lys Ile Trp Leu Asp Ser Glu
225 230 235 240
Glu Gly Gln Asn Ala Ile Lys Lys Ala Phe Asn Ser Thr Ser Arg Phe
245 250 255
Ala Arg Leu Gln His Ile Gln Thr Cys Leu Ala Gly Asp Leu Leu Phe
260 265 270
Met Arg Phe Arg Thr Thr Thr Gly Asp Ala Met Gly Met Asn Met Ile
275 280 285
Ser Lys Gly Val Glu Tyr Ser Leu Lys Gln Met Val Glu Glu Tyr Gly
290 295 300
Trp Glu Asp Met Glu Val Val Ser Val Ser Gly Asn Tyr Cys Thr Asp
305 310 315 320
Lys Lys Pro Ala Ala Ile Asn Trp Ile Glu Gly Arg Gly Lys Ser Val
325 330 335
Val Ala Glu Ala Thr Ile Pro Gly Asp Val Val Arg Lys Val Leu Lys
340 345 350
Ser Asp Val Ser Ala Leu Val Glu Leu Asn Ile Ala Lys Asn Leu Val
355 360 365
Gly Ser Ala Met Ala Gly Ser Val Gly Gly Phe Asn Ala His Ala Ala
370 375 380
Asn Leu Val Thr Ala Val Phe Leu Ala Leu Gly Gln Asp Pro Ala Gln
385 390 395 400
Asn Val Glu Ser Ser Asn Cys Ile Thr Leu Met Lys Glu Val Asp Gly
405 410 415
Asp Leu Arg Ile Ser Val Ser Met Pro Ser Ile Glu Val Gly Thr Ile
420 425 430
Gly Gly Gly Thr Val Leu Glu Pro Gln Gly Ala Met Leu Asp Leu Leu
435 440 445
Gly Val Arg Gly Pro His Ala Thr Ala Pro Gly Thr Asn Ala Arg Gln
450 455 460
Leu Ala Arg Ile Val Ala Cys Ala Val Leu Ala Gly Glu Leu Ser Leu
465 470 475 480
Cys Ala Ala Leu Ala Ala Gly His Leu Val Gln Ser His Met Thr His
485 490 495
Asn Arg Lys Pro Ala Glu Pro Thr Lys Pro Asn Asn Leu Asp Ala Thr
500 505 510
Asp Ile Asn Arg Leu Lys Asp Gly Ser Val Thr Cys Ile Lys Ser
515 520 525
<210> 28
<211> 1581
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 28
atggcggctg atcagctcgt taaaacggaa gtaacaaaga aatccttcac cgctcccgta 60
cagaaagctt ctactcctgt cttgaccaac aagaccgtta tctccggctc taaagtgaaa 120
tcgctctctt ccgcccagag cagctcatcc ggtccgtcgt cttcctccga agaggacgac 180
tcacgtgata ttgaatcact ggataaaaag attcgaccgc tcgaagagct tgaggcgctt 240
ttgagctccg gcaataccaa acagttgaag aacaaggagg ttgccgccct tgtgattcac 300
ggcaaacttc cgctctatgc cttggagaag aagttgggag acaccacgcg cgcagtggcc 360
gtgcgccgca aggcactgtc tattctcgcg gaggcacccg tcctggcttc cgaccgcttg 420
ccctacaaga actacgatta cgaccgtgtc ttcggcgcat gttgcgagaa tgtaatcgga 480
tatatgcccc ttcccgttgg cgtgatcggc ccacttgtca tcgatggcac atcataccat 540
attccaatgg cgaccaccga gggttgcctc gtggcgtcgg ccatgcgcgg ttgtaaggcc 600
atcaacgccg gaggtggagc aactaccgta cttactaagg acggcatgac tcgtggcccc 660
gtggtgcgtt tcccgactct taaacgctcc ggtgcgtgca agatctggtt ggattccgag 720
gagggacaga acgcaatcaa aaaagctttc aacagcactt ctcgcttcgc tcgacttcag 780
cacattcaga cctgtctcgc tggtgacttg ttgttcatgc gttttcgcac caccactgga 840
gacgcgatgg gcatgaatat gattagcaaa ggagtggaat actccctcaa acaaatggtg 900
gaagaatatg gctgggaaga catggaagtg gtttccgtgt ctggtaatta ttgtaccgac 960
aagaagccgg cagccatcaa ctggattgag ggccgtggca agtccgtggt ggcggaggcg 1020
acaattccgg gtgatgtcgt gcgcaaagtt ctcaagtctg acgtgtcagc gttggtagaa 1080
ctgaacatcg caaaaaacct tgtaggatcg gcaatggctg gttcagtggg cggttttaat 1140
gcgcacgcag ccaacctcgt caccgctgtg ttccttgccc tgggtcagga tccagcacag 1200
aatgtggaaa gctcgaactg catcaccctg atgaaagaag tggatggcga cttgcgtatc 1260
tccgtctcga tgccatcaat cgaagtgggt accatcggtg gcggaaccgt gttggaacca 1320
cagggagcaa tgcttgatct cctcggcgtt cgaggtccgc acgccacggc gcccggcacc 1380
aacgcacgtc agctggcacg catcgtggct tgcgcggttc tcgcgggtga attgagcctc 1440
tgcgcagccc tggcagcggg tcacctggtg caatctcaca tgacacataa ccgtaaacca 1500
gccgagccca ccaagccgaa taacctcgac gccactgata ttaaccgctt gaaggatggt 1560
tccgtcacct gcatcaaatc t 1581
<210> 29
<211> 425
<212> PRT
<213> 金黄色葡萄球菌（Staphylococcus aureus）
<400> 29
Met Gln Ser Leu Asp Lys Asn Phe Arg His Leu Ser Arg Gln Gln Lys
1 5 10 15
Leu Gln Gln Leu Val Asp Lys Gln Trp Leu Ser Glu Asp Gln Phe Asp
20 25 30
Ile Leu Leu Asn His Pro Leu Ile Asp Glu Glu Val Ala Asn Ser Leu
35 40 45
Ile Glu Asn Val Ile Ala Gln Gly Ala Leu Pro Val Gly Leu Leu Pro
50 55 60
Asn Ile Ile Val Asp Asp Lys Ala Tyr Val Val Pro Met Met Val Glu
65 70 75 80
Glu Pro Ser Val Val Ala Ala Ala Ser Tyr Gly Ala Lys Leu Val Asn
85 90 95
Gln Thr Gly Gly Phe Lys Thr Val Ser Ser Glu Arg Ile Met Ile Gly
100 105 110
Gln Ile Val Phe Asp Gly Val Asp Asp Thr Glu Lys Leu Ser Ala Asp
115 120 125
Ile Lys Ala Leu Glu Lys Gln Ile His Lys Ile Ala Asp Glu Ala Tyr
130 135 140
Pro Ser Ile Lys Ala Arg Gly Gly Gly Tyr Gln Arg Ile Ala Ile Asp
145 150 155 160
Thr Phe Pro Glu Gln Gln Leu Leu Ser Leu Lys Val Phe Val Asp Thr
165 170 175
Lys Asp Ala Met Gly Ala Asn Met Leu Asn Thr Ile Leu Glu Ala Ile
180 185 190
Thr Ala Phe Leu Lys Asn Glu Ser Pro Gln Ser Asp Ile Leu Met Ser
195 200 205
Ile Leu Ser Asn His Ala Thr Ala Ser Val Val Lys Val Gln Gly Glu
210 215 220
Ile Asp Val Lys Asp Leu Ala Arg Gly Glu Arg Thr Gly Glu Glu Val
225 230 235 240
Ala Lys Arg Met Glu Arg Ala Ser Val Leu Ala Gln Val Asp Ile His
245 250 255
Arg Ala Ala Thr His Asn Lys Gly Val Met Asn Gly Ile His Ala Val
260 265 270
Val Leu Ala Thr Gly Asn Asp Thr Arg Gly Ala Glu Ala Ser Ala His
275 280 285
Ala Tyr Ala Ser Arg Asp Gly Gln Tyr Arg Gly Ile Ala Thr Trp Arg
290 295 300
Tyr Asp Gln Lys Arg Gln Arg Leu Ile Gly Thr Ile Glu Val Pro Met
305 310 315 320
Thr Leu Ala Ile Val Gly Gly Gly Thr Lys Val Leu Pro Ile Ala Lys
325 330 335
Ala Ser Leu Glu Leu Leu Asn Val Asp Ser Ala Gln Glu Leu Gly His
340 345 350
Val Val Ala Ala Val Gly Leu Ala Gln Asn Phe Ala Ala Cys Arg Ala
355 360 365
Leu Val Ser Glu Gly Ile Gln Gln Gly His Met Ser Leu Gln Tyr Lys
370 375 380
Ser Leu Ala Ile Val Val Gly Ala Lys Gly Asp Glu Ile Ala Gln Val
385 390 395 400
Ala Glu Ala Leu Lys Gln Glu Pro Arg Ala Asn Thr Gln Val Ala Glu
405 410 415
Arg Ile Leu Gln Glu Ile Arg Gln Gln
420 425
<210> 30
<211> 1275
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 30
atgcaatccc tcgataaaaa tttccgtcac ctttctcgac agcagaaatt gcagcaactt 60
gttgacaagc aatggctctc ggaggatcaa ttcgatatcc tgctcaatca cccactgatc 120
gatgaggagg tcgccaattc cctgattgag aacgtcatcg cgcagggtgc actgcccgtg 180
ggccttctgc ctaacatcat cgtcgatgac aaggcctacg tagtacctat gatggtcgaa 240
gaaccgtccg tggtggcagc tgcttcctat ggtgccaagc tggtcaacca gaccggcgga 300
ttcaagactg tgtcttctga acgcattatg atcggacaga ttgtcttcga tggtgttgat 360
gatacggaaa agctctctgc agacatcaag gcattggaga aacagatcca taaaatcgcg 420
gatgaggcgt acccctccat caaagcgcgc ggcggcggct atcagcgaat cgctattgac 480
accttccctg aacagcagct gctctccctg aaagtcttcg tggatactaa ggacgccatg 540
ggtgcaaata tgttgaacac catcctggag gccatcaccg cgttcctgaa gaacgagagc 600
cctcagtcgg atattttgat gtcaatcctg tccaatcacg ccactgcatc tgtggtaaaa 660
gtgcaaggtg aaatcgatgt caaagatctc gcccgcggag aacgcacggg cgaagaggtc 720
gcaaaacgaa tggagcgcgc ctctgttctt gcgcaggtgg atatccaccg cgctgcgacc 780
cacaataaag gtgtgatgaa cggcatccat gcagtcgtac ttgccaccgg caacgatact 840
cgcggtgcag aagcgtctgc ccatgcatat gcgtcgcgag atggtcaata ccgtggcatt 900
gctacatggc gttacgatca aaaacgccag cgtctcattg gtacgatcga ggttcctatg 960
accctggcca tcgtcggcgg aggcaccaaa gtcctcccga tcgcaaaggc ctcgctcgaa 1020
cttttgaatg tcgatagcgc acaagaactc ggtcacgtcg tagcagccgt gggtctggcg 1080
cagaacttcg cagcgtgtcg agcacttgtc tccgagggca ttcagcaggg tcacatgtcc 1140
cttcagtaca aaagcttggc aattgttgtg ggtgcaaagg gtgatgagat tgcccaggtc 1200
gcagaagcac tcaaacaaga gccacgtgcg aacacgcagg ttgccgagcg tatccttcaa 1260
gaaattcgac aacaa 1275
<210> 31
<211> 446
<212> PRT
<213> Candidatus Halobonum tyrrellensis G22
<400> 31
Met Ser Asp Ala Asp Thr Glu Arg Leu Ala Ala Arg Val Arg Glu Gly
1 5 10 15
Asp Leu Arg Leu Tyr Glu Leu Glu Glu His Ala Asp Ala Asp Thr Ala
20 25 30
Val Ala Ala Arg Arg Leu Leu Val Glu Glu Ala Ser Gly Ala Glu Leu
35 40 45
Glu Thr Thr Gly Glu Tyr Ala Phe Pro Ala Glu Met Ala Thr Gly Thr
50 55 60
Asn Val Glu Asn Met Val Gly Ala Val Gln Val Pro Val Gly Val Val
65 70 75 80
Gly Pro Val Thr Ile Asp Gly Gly Ala Leu Ser Gly Glu Arg His Val
85 90 95
Pro Met Ala Thr Thr Glu Gly Ala Leu Leu Ala Ser Val Asn Arg Gly
100 105 110
Cys Ser Ala Leu Asn Thr Ala Gly Gly Ala Ser Ala Arg Val Leu Lys
115 120 125
Asn Arg Met Thr Arg Ala Pro Val Phe Arg Val Asp Gly Val Val Glu
130 135 140
Ala Glu Ala Leu Ala Ser Trp Thr Arg Asp Asn Val Glu Ala Leu Arg
145 150 155 160
Ala Val Ala Glu Ala Thr Thr Asn His Gly Glu Leu Arg Glu Val Arg
165 170 175
Pro Tyr Ala Val Gly Asn Asn Val Tyr Leu Arg Phe Ala Tyr Asp Thr
180 185 190
Lys Asp Ala Met Gly Met Asn Met Ala Thr Ile Ala Thr Glu Ala Ala
195 200 205
Cys Glu Val Val Glu Asp Glu Thr Gly Ala Glu Leu Val Ala Leu Ser
210 215 220
Gly Asn Leu Cys Ser Asp Lys Lys Pro Ala Ala Val Asn Ala Val Glu
225 230 235 240
Gly Arg Gly Arg Thr Val Ala Ala Asp Val Thr Val Pro Arg Glu Thr
245 250 255
Val Arg Glu Thr Leu Lys Thr Thr Pro Glu Ala Val Ala Glu Val Asn
260 265 270
Thr Arg Lys Asn Leu Val Gly Ser Ala Lys Ala Gly Ser Leu Gly Phe
275 280 285
Asn Ala His Val Ala Asn Ala Val Ala Ala Val Phe Leu Ala Thr Gly
290 295 300
Gln Asp Ala Ala Gln Val Val Glu Gly Ala Asn Ala Ile Thr Thr Ala
305 310 315 320
Glu Val Val Ala Gly Asp Ala Pro Ser Arg Thr Val Asp Arg Asn Gly
325 330 335
Ala Ala Glu Ser Gly Val Gly Pro Asp Gly Asp Ala Pro Asp Gly Asp
340 345 350
Ala Pro Asp Gly Asp Gly Ala Ala Ala Gly Arg Gly Gly Asp Leu Tyr
355 360 365
Val Ser Val Thr Leu Ala Ser Leu Glu Val Gly Thr Val Gly Gly Gly
370 375 380
Thr Lys Leu Pro Thr Gln Ala Glu Ala Leu Asp Val Leu Gly Val Arg
385 390 395 400
Gly Gly Gly Asp Pro Ala Gly Ser Asn Ala Asp Ala Leu Ala Glu Ala
405 410 415
Val Ala Val Thr Ala Leu Ala Gly Glu Leu Ser Leu Leu Ser Ala Leu
420 425 430
Ala Ser Arg Asn Leu Ser Ser Ala His Ala Glu Leu Gly Arg
435 440 445
<210> 32
<211> 1338
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 32
atgtcagacg ctgacaccga gcgtctggct gcgcgcgttc gcgagggtga tctgcgcctg 60
tatgagctgg aggagcacgc ggatgcagat actgctgtcg ccgctcgtcg tctcttggtg 120
gaagaggcct ccggtgcaga acttgagact accggtgaat atgctttccc tgcagaaatg 180
gcaaccggca ccaatgtcga gaacatggtc ggtgctgttc aggttccagt gggcgtagtg 240
ggtcctgtta cgattgatgg cggtgcgctg tctggcgagc gtcacgtccc aatggctaca 300
accgaaggtg cgctcctcgc gtccgtcaat cgcggctgct cggccctcaa cacagctggt 360
ggtgcatcag ctcgtgtcct gaagaatcgc atgactcgtg cccccgtctt tcgcgtcgat 420
ggcgtcgtcg aagccgaggc gcttgcttcg tggacccgcg acaacgtgga agcgctccgt 480
gcagtggctg aagctaccac caaccacgga gaactccgcg aagtgcgccc atacgctgtg 540
ggcaacaacg tctaccttcg attcgcttat gataccaagg atgcaatggg catgaatatg 600
gccactatcg ccaccgaggc ggcgtgcgag gtggtggaag atgaaaccgg cgctgagctg 660
gttgcactga gcggtaacct ttgctctgat aagaagccag ccgctgtgaa cgcagtggaa 720
ggtcgcggtc gcacagttgc tgctgatgtg acggttcctc gcgagaccgt tcgcgagacc 780
ctgaagacca cccctgaggc cgtggccgag gttaacacgc gcaagaactt ggtgggaagc 840
gcgaaggccg gctcattggg atttaatgct catgtggcaa acgccgtggc agcagtcttt 900
cttgcaacag gtcaggatgc tgcgcaggtc gtggaaggtg ctaacgccat cactacggcg 960
gaagtcgtcg caggcgatgc tccatcccgt accgtggatc gcaatggcgc tgctgaatcc 1020
ggcgtaggtc cggatggtga tgcacctgac ggtgatgcac cggatggtga cggcgcagca 1080
gcaggtcgtg gcggcgacct ctatgttagc gtgaccctgg cctcactgga ggtcggtacc 1140
gtgggcggcg gtacgaaact gcccacccaa gcggaagcat tggacgtgct cggtgtacgt 1200
ggaggaggtg atcctgctgg tagcaacgca gatgctctcg ctgaagccgt tgctgtgacc 1260
gctttggctg gtgagttgtc tcttctttcc gctctcgcca gccgtaacct ctcgtcggca 1320
cacgctgaac tgggacgc 1338
<210> 33
<211> 665
<212> PRT
<213> 黄瓜（Cucumis sativus）（黄瓜）
<400> 33
Met Thr His Cys Ser Gln Leu Pro Thr His Lys Ala Thr Thr Thr Glu
1 5 10 15
Lys Asn Asp Glu Pro Ile Pro Leu Gly Pro Val Gln Leu Ser Trp Pro
20 25 30
Gly Ser Val Val Glu Lys Gly Glu Lys Ser Ser Phe His Tyr Lys Pro
35 40 45
Pro Phe Ala Ser Arg Ser Gly Ala Gly Val Phe His Phe Leu Phe Phe
50 55 60
Ser Cys Leu Thr Met Asp Ala Arg Arg Arg Arg Ser Ser Thr Leu Val
65 70 75 80
Lys Lys Leu Thr Ala Asp Glu Pro Pro Val Lys Ser Met Asp Lys Asn
85 90 95
Pro Leu Lys Val Lys Met Leu Glu Arg Gln His Val His Asp Asp Ala
100 105 110
Val Lys Ala Ser Asp Val Leu Pro Leu Pro Ile Tyr Leu Thr Asn Ala
115 120 125
Ala Phe Phe Thr Leu Phe Phe Ser Val Val Tyr Phe Leu Leu Thr Arg
130 135 140
Trp Arg Glu Lys Ile Arg Ser Ser Thr Pro Leu His Val Val Thr Leu
145 150 155 160
Ser Glu Met Val Ala Ile Ser Ala Phe Ile Ala Ser Phe Ile Tyr Leu
165 170 175
Leu Gly Phe Phe Gly Ile Asp Phe Val Gln Ser Ile Phe Arg Pro Ser
180 185 190
His Asp Val Trp Thr Ser Glu Asp Asp Glu Val Val Ile Ile Lys Glu
195 200 205
Asp Thr Arg Lys Val Pro Cys Gly Ala Gly Ile Asp Cys Ser Ile Pro
210 215 220
Ile Leu Ala Pro Pro Met Pro Ser Val Pro Lys Val Val Asp Pro Leu
225 230 235 240
Pro Val Ser Ile Asp Leu Thr Glu Glu Asp Glu Glu Ile Val Lys Ser
245 250 255
Val Val Asp Gly Ser Thr Pro Ser Tyr Ser Leu Glu Ser Lys Leu Gly
260 265 270
Asp Cys Gly Arg Ala Ala Ala Ile Arg Arg Val Ala Leu Gln Arg Val
275 280 285
Thr Gly Lys Ser Leu Ser Gly Leu Pro Leu Glu Gly Phe Asp Tyr Ala
290 295 300
Ser Ile Leu Gly Gln Cys Cys Glu Met Pro Ile Gly Tyr Val Gln Ile
305 310 315 320
Pro Val Gly Ile Ala Gly Pro Leu Leu Leu Asp Gly Lys Glu Tyr Ser
325 330 335
Val Pro Met Ala Thr Thr Glu Gly Cys Leu Val Ala Ser Thr Asn Arg
340 345 350
Gly Cys Lys Ala Ile Met Ile Ser Gly Gly Ala Asn Ser Val Leu Leu
355 360 365
Arg Asp Ala Met Thr Arg Ala Pro Val Met Arg Phe Ala Thr Ala Lys
370 375 380
Arg Ala Ala Glu Leu Lys Phe Tyr Val Glu Asp Pro Ala Asn Phe Asp
385 390 395 400
Thr Leu Ala Ser Val Phe Asn Lys Ser Ser Arg Phe Gly Arg Leu Gln
405 410 415
Ser Ile Lys Cys Ala Ile Ala Gly Lys Asn Leu Tyr Met Arg Phe Ser
420 425 430
Cys Ser Thr Gly Asp Ala Met Gly Met Asn Met Val Ser Lys Gly Val
435 440 445
Gln Asn Val Leu Asp Phe Leu Gln Asn Asp Phe Pro Asp Met Asp Val
450 455 460
Ile Gly Ile Ser Gly Asn Tyr Cys Ser Asp Lys Lys Pro Ala Ala Val
465 470 475 480
Asn Trp Ile Glu Gly Arg Gly Lys Ser Val Val Cys Glu Val Thr Ile
485 490 495
Lys Gly Asp Val Val Arg Lys Val Leu Lys Thr Asp Val Gln Ala Leu
500 505 510
Val Glu Leu Asn Met Leu Lys Asn Leu Thr Gly Ser Ala Met Ala Gly
515 520 525
Ala Leu Gly Gly Phe Asn Ala His Ala Ser Asn Ile Val Ser Ala Ile
530 535 540
Tyr Ile Ala Thr Gly Gln Asp Pro Ala Gln Asn Val Glu Ser Ser His
545 550 555 560
Cys Ile Thr Met Met Glu Ala Val Asn Asp Gly Gln Asp Leu His Val
565 570 575
Ser Val Thr Met Pro Ser Ile Glu Val Gly Thr Val Gly Gly Gly Thr
580 585 590
Gln Leu Ala Ser Gln Ser Ala Cys Leu Asn Leu Leu Gly Val Lys Gly
595 600 605
Ala Asn Arg Glu Ala Pro Gly Ser Asn Ser Arg Leu Leu Ala Thr Ile
610 615 620
Val Ala Ala Ser Val Leu Ala Gly Glu Leu Ser Leu Met Ser Ala Ile
625 630 635 640
Ser Ala Gly Gln Leu Val Arg Ser His Met Lys Tyr Asn Arg Ser Ser
645 650 655
Arg Asp Ile Thr Lys Ala Ser Ser Ser
660 665
<210> 34
<211> 1995
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 34
atgacccatt gctcccagtt gccgacccac aaggcgacga cgactgaaaa aaacgatgag 60
cctattcccc ttggccccgt gcaactgagc tggccgggtt ccgttgttga aaagggagaa 120
aagtcatcgt tccattataa gcccccattc gcgtctcgca gcggcgcagg cgtatttcat 180
tttctctttt tctcgtgctt gactatggac gcccgtcgcc gtcgctcttc gactctggtg 240
aagaaactta ctgcggatga gcctccagta aaatctatgg ataaaaatcc tcttaaagtc 300
aagatgcttg aacgacagca tgtccatgac gatgccgtaa aggcgtcgga tgtgctccca 360
ctgccaatct accttactaa cgctgcattc ttcactctgt tcttttccgt ggtatatttc 420
cttcttacgc gctggcgtga gaagattcgt agctccacgc ccctgcacgt agtaaccctc 480
tcagaaatgg tggctatctc tgccttcatc gcctcgttca tttacctgct tggctttttc 540
ggcattgatt tcgtgcagtc catcttccgc ccctcccacg atgtctggac ctcagaggac 600
gacgaagtgg tcatcatcaa agaagacact cgcaaggtcc catgtggcgc cggaattgac 660
tgttccattc caattctggc ccctcctatg ccctcggtgc cgaaggtggt tgatccgctg 720
cctgtgtcca ttgatctcac cgaggaggat gaagagattg tgaagtcagt cgtcgacggc 780
tcgacacctt cttactcatt ggagtctaag ctcggagatt gtggccgagc ggctgctatc 840
cgacgtgttg cacttcagcg cgtgactggc aagtcgctgt ctggcctccc gctggagggc 900
ttcgactatg cttccatcct cggacagtgt tgcgagatgc ccattggcta cgtgcaaatt 960
cccgtgggaa ttgccggccc attgcttttg gatggtaagg aatacagcgt ccccatggcg 1020
acgaccgaag gatgccttgt ggcctccacc aaccgaggct gcaaggcgat catgatctcc 1080
ggaggcgcta actccgtcct ccttcgtgat gccatgacac gtgcgccagt gatgcgcttt 1140
gcgactgcta agcgcgccgc ggaactcaaa ttctacgttg aagacccggc taacttcgat 1200
acccttgcct ccgtatttaa caaatctagc cgctttggac gcctgcagag catcaaatgt 1260
gccatcgccg gcaaaaacct ctatatgcga ttttcgtgct ccactggcga cgctatggga 1320
atgaatatgg tttccaaggg cgtacagaac gttctggact ttctccagaa tgattttcca 1380
gacatggatg taatcggaat cagcggaaac tattgttcag ataagaagcc agccgccgtt 1440
aattggatcg aaggacgcgg caagtccgtc gtgtgcgagg ttaccatcaa aggcgatgtt 1500
gtccgcaagg tcctgaagac agatgtccag gcactggtgg aactcaacat gttgaaaaat 1560
cttacgggtt ccgcaatggc tggcgcactt ggtggcttta acgctcatgc atccaacatc 1620
gtttcagcca tctacatcgc gacgggacag gatcccgctc aaaacgtgga aagctcccat 1680
tgcatcacta tgatggaggc cgtaaacgat ggtcaggatc ttcacgtgag cgtgaccatg 1740
ccatccatcg aggtgggaac cgtaggcggt ggcacgcaac tggcatccca gtccgcctgc 1800
ctgaacctct tgggtgttaa gggcgcgaat cgcgaagctc caggttcaaa ttctcgcttg 1860
ctcgccacca ttgtggccgc atcagttctt gcaggcgaac ttagcctcat gtcagcgatc 1920
tccgccggcc aactcgtccg cagccacatg aaatacaatc gttcatcacg cgatatcacc 1980
aaggcttcat catcc 1995
<210> 35
<211> 428
<212> PRT
<213> Lokiarchaeum sp. GC14_75
<400> 35
Met Ile Met Lys Asp Phe His Ser Asp Ile Ser Gly Phe Tyr Lys Leu
1 5 10 15
Ser Ile Asp Glu Arg Gln Lys Leu Leu Ser Lys Leu Val Asn Leu Asn
20 25 30
Pro Glu Asp Leu Glu Ile Leu Lys Glu Leu Gly Tyr Phe Thr Pro Thr
35 40 45
Gln Ile Asp Thr Leu Ile Glu Asn Val Val Gly Ser Tyr Gln Leu Pro
50 55 60
Leu Gly Leu Ala Phe Asn Phe Arg Ile Asn Ser Lys Asp Tyr Ile Ile
65 70 75 80
Pro Met Val Ile Glu Glu Pro Ser Val Val Ala Ala Ala Ser Asn Ala
85 90 95
Ala Lys Met Ala Arg Lys His Gly Gly Phe His Ser Glu Asp Val Pro
100 105 110
Pro Ile Met Ile Ser Gln Ile Gln Ile Thr Lys Leu Lys Asp Ile Glu
115 120 125
Ala Ala Lys Asp Val Leu Glu Ala Lys Lys Gln Asp Leu Leu Lys Ile
130 135 140
Ala Asn Glu Gln Asp Pro Leu Leu Asn Lys Leu Gly Gly Gly Ala Gln
145 150 155 160
Asn Ile Glu Ile His Glu Ile Gln Thr Asn Lys Gly Lys Met Ile Ile
165 170 175
Leu His Leu Leu Val Asn Val Leu Asp Ala Met Gly Ala Asn Ile Val
180 185 190
Asn Thr Met Ala Glu Ala Ile Ser Pro Tyr Ile Glu Glu Ile Cys Gly
195 200 205
Gly Lys Ile Tyr Leu Arg Ile Val Ser Asn Leu Ala Thr His Arg Val
210 215 220
Ala Arg Ser Lys Ala Thr Phe Asp Lys Glu Met Leu Gly Gly Glu Glu
225 230 235 240
Val Val Glu Gly Ile Met Asn Ala Tyr Glu Phe Ala Leu Ala Asp Pro
245 250 255
Tyr Arg Ala Thr Thr His Asn Lys Gly Ile Met Asn Gly Val Val Ala
260 265 270
Leu Thr Leu Ala Thr Gly Asn Asp Thr Arg Ala Ile Glu Ser Gly Ala
275 280 285
His Ser Tyr Ala Ser Leu Ser Gly Arg Tyr Leu Pro Leu Thr Lys Phe
290 295 300
Asp Val Asp Ser Gln Gly Asn Leu Val Gly Glu Ile Glu Ile Pro Leu
305 310 315 320
Ala Leu Gly Ile Ile Gly Gly Met Thr Lys Ile His Pro Met Ala Arg
325 330 335
Leu Ala Leu Lys Ile Leu Asn Leu Lys Ser Ser Ser Glu Leu Ala Gln
340 345 350
Val Gly Ala Ala Leu Gly Leu Ala Gln Asn Val Ala Ala Leu Arg Ala
355 360 365
Leu Ala Ala Glu Gly Ile Gln Lys Gly His Met Ala Leu His Ser Arg
370 375 380
Asn Ile Ala Lys Val Ala Gly Val Pro Asp Glu Leu Ile Glu Lys Val
385 390 395 400
Ser Lys Lys Met Val Ala Asp Lys Lys Ile Arg Val Asp Tyr Ala Lys
405 410 415
Glu Ile Phe Gln Lys Ile Lys Asp Gly Glu Asn Ile
420 425
<210> 36
<211> 1284
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 36
atgatcatga aagacttcca ctccgatatt tccggtttct acaagctgtc catcgatgag 60
cgccagaagc ttttgagcaa gcttgtaaac ctgaacccgg aagatctgga aattcttaag 120
gaactgggct atttcacccc gactcaaatt gacaccctga tcgagaacgt tgtcggctcc 180
tatcaacttc ccttgggatt ggccttcaac tttcgcatca attctaagga ttacatcatc 240
cctatggtga tcgaagagcc atccgtggta gctgcagcat ccaatgcggc taagatggct 300
cgcaaacacg gcggttttca ctctgaagat gttcctccaa tcatgatctc acagattcaa 360
atcaccaaat tgaaagatat cgaggcagct aaagatgtcc tcgaggcgaa gaagcaggat 420
ttgctcaaga ttgccaacga acaagatccg ctgctgaaca aacttggcgg tggcgcccag 480
aacatcgaaa tccacgaaat ccaaactaac aagggcaaga tgatcatcct ccatctgctc 540
gtaaacgttc tcgatgccat gggtgccaac atcgtcaata ccatggcgga ggctatctct 600
ccatacatcg aagagatctg cggtggcaag atctaccttc gcattgtttc caacctggca 660
acccaccgtg tggcgcgttc caaggcaacc ttcgataagg aaatgctggg cggcgaagag 720
gtggtggaag gcatcatgaa tgcatatgaa ttcgcgctgg cggaccccta ccgcgctacc 780
acccataaca agggtatcat gaatggtgtt gtggctttga ctctggcgac gggtaatgac 840
acccgcgcaa tcgagagcgg agcccactct tacgcatccc tttccggccg ttacctgccc 900
ctgaccaagt ttgacgtaga ttctcaaggt aaccttgtgg gagaaatcga gattcctctt 960
gccctgggca ttatcggtgg tatgaccaag atccatccaa tggcccgtct ggctttgaag 1020
attcttaacc ttaaatcatc ttcagaactt gcacaggtag gtgcggcgct gggacttgcc 1080
caaaacgttg cagctctgcg cgcgctcgcc gctgaaggta tccagaaagg tcacatggca 1140
ctccactcac gtaatattgc aaaggtcgca ggcgtccctg acgaactgat tgaaaaagtg 1200
tctaaaaaga tggtcgccga taaaaagatt cgtgtggatt atgcaaagga aatcttccag 1260
aagatcaagg acggcgaaaa catt 1284
<210> 37
<211> 931
<212> PRT
<213> 中国仓鼠（Cricetulus griseus）
<400> 37
Met Thr Pro Asp Met Arg Lys Thr Gly Arg Cys Glu Ala Phe Leu Glu
1 5 10 15
Gln Arg Ser Ile Leu Leu Pro Asn Arg Phe Leu Phe Ser Gly His Pro
20 25 30
His Ala Ala Thr Leu Thr Leu Pro Gln Glu Gly Pro Ser Gln Val Arg
35 40 45
Phe Arg Gly Arg Gly Thr Pro Pro Ala Gly Ser Pro Ile Ala Arg Glu
50 55 60
Gly Gly Val Leu Ala Gly Ala Pro His Gly Ser Gly Thr Asn Lys Lys
65 70 75 80
Val Val Met Leu Glu Leu Asp Gln Leu Leu Val Gly Ser Ala Val Val
85 90 95
Arg Asp Gly Ala Val Pro Val Leu Arg Pro Gly Ser Arg Asp Arg Val
100 105 110
Ala Thr Met Leu Ser Arg Leu Phe Arg Met His Gly Leu Phe Val Ala
115 120 125
Ser His Pro Trp Glu Val Ile Val Gly Thr Val Thr Leu Thr Ile Cys
130 135 140
Met Met Ser Met Asn Met Phe Thr Gly Asn Asn Lys Ile Cys Gly Trp
145 150 155 160
Asn Tyr Glu Cys Pro Lys Phe Glu Glu Asp Val Leu Ser Ser Asp Ile
165 170 175
Ile Ile Leu Thr Ile Thr Arg Cys Ile Ala Ile Leu Tyr Ile Tyr Phe
180 185 190
Gln Phe Gln Asn Leu Arg Gln Leu Gly Ser Lys Tyr Ile Leu Gly Ile
195 200 205
Ala Gly Leu Phe Thr Ile Phe Ser Ser Phe Val Phe Ser Thr Val Val
210 215 220
Ile His Phe Leu Asp Lys Glu Leu Thr Gly Leu Asn Glu Ala Leu Pro
225 230 235 240
Phe Phe Leu Leu Leu Ile Asp Leu Ser Arg Ala Ser Ala Leu Ala Lys
245 250 255
Phe Ala Leu Ser Ser Asn Ser Gln Asp Glu Val Arg Glu Asn Ile Ala
260 265 270
Arg Gly Met Ala Ile Leu Gly Pro Thr Phe Thr Leu Asp Ala Leu Val
275 280 285
Glu Cys Leu Val Ile Gly Val Gly Thr Met Ser Gly Val Arg Gln Leu
290 295 300
Glu Ile Met Cys Cys Phe Gly Cys Met Ser Val Leu Ala Asn Tyr Phe
305 310 315 320
Val Phe Met Thr Phe Phe Pro Ala Cys Val Ser Leu Val Leu Glu Leu
325 330 335
Ser Arg Glu Ser Arg Glu Gly Arg Pro Ile Trp Gln Leu Ser His Phe
340 345 350
Ala Arg Val Leu Glu Glu Glu Glu Asn Lys Pro Asn Pro Val Thr Gln
355 360 365
Arg Val Lys Met Ile Met Ser Leu Gly Leu Val Leu Val His Ala His
370 375 380
Ser Arg Trp Ile Ala Asp Pro Ser Pro Gln Asn Ser Thr Thr Glu His
385 390 395 400
Ser Lys Val Ser Leu Gly Leu Asp Glu Asp Val Ser Lys Arg Ile Glu
405 410 415
Pro Ser Val Ser Leu Trp Gln Phe Tyr Leu Ser Lys Met Ile Ser Met
420 425 430
Asp Ile Glu Gln Val Val Thr Leu Ser Leu Ala Phe Leu Leu Ala Val
435 440 445
Lys Tyr Ile Phe Phe Glu Gln Ala Glu Thr Glu Ser Thr Leu Ser Leu
450 455 460
Lys Asn Pro Ile Thr Ser Pro Val Val Thr Pro Lys Lys Ala Pro Asp
465 470 475 480
Asn Cys Cys Arg Arg Glu Pro Leu Leu Val Arg Arg Ser Glu Lys Leu
485 490 495
Ser Ser Val Glu Glu Glu Pro Gly Val Ser Gln Asp Arg Lys Val Glu
500 505 510
Val Ile Lys Pro Leu Val Val Glu Thr Glu Ser Ala Ser Arg Ala Thr
515 520 525
Phe Val Leu Gly Ala Ser Gly Thr Ser Pro Pro Val Ala Ala Arg Thr
530 535 540
Gln Glu Leu Glu Ile Glu Leu Pro Ser Glu Pro Arg Pro Asn Glu Glu
545 550 555 560
Cys Leu Gln Ile Leu Glu Ser Ala Glu Lys Gly Ala Lys Phe Leu Ser
565 570 575
Asp Ala Glu Ile Ile Gln Leu Val Asn Ala Lys His Ile Pro Ala Tyr
580 585 590
Lys Leu Glu Thr Leu Met Glu Thr His Glu Arg Gly Val Ser Ile Arg
595 600 605
Arg Gln Leu Leu Ser Thr Lys Leu Pro Glu Pro Ser Ser Leu Gln Tyr
610 615 620
Leu Pro Tyr Arg Asp Tyr Asn Tyr Ser Leu Val Met Gly Ala Cys Cys
625 630 635 640
Glu Asn Val Ile Gly Tyr Met Pro Ile Pro Val Gly Val Ala Gly Pro
645 650 655
Leu Cys Leu Asp Gly Lys Glu Tyr Gln Val Pro Met Ala Thr Thr Glu
660 665 670
Gly Cys Leu Val Ala Ser Thr Asn Arg Gly Cys Arg Ala Ile Gly Leu
675 680 685
Gly Gly Gly Ala Ser Ser Arg Val Leu Ala Asp Gly Met Thr Arg Gly
690 695 700
Pro Val Val Arg Leu Pro Arg Ala Cys Asp Ser Ala Glu Val Lys Ala
705 710 715 720
Trp Leu Glu Thr Pro Glu Gly Phe Ala Val Ile Lys Asp Ala Phe Asp
725 730 735
Ser Thr Ser Arg Phe Ala Arg Leu Gln Lys Leu His Val Thr Met Ala
740 745 750
Gly Arg Asn Leu Tyr Ile Arg Phe Gln Ser Lys Thr Gly Asp Ala Met
755 760 765
Gly Met Asn Met Ile Ser Lys Gly Thr Glu Lys Ala Leu Leu Lys Leu
770 775 780
Gln Glu Phe Phe Pro Glu Met Gln Ile Leu Ala Val Ser Gly Asn Tyr
785 790 795 800
Cys Thr Asp Lys Lys Pro Ala Ala Ile Asn Trp Ile Glu Gly Arg Gly
805 810 815
Lys Thr Val Val Cys Glu Ala Val Ile Pro Ala Lys Val Val Arg Glu
820 825 830
Val Leu Lys Thr Thr Thr Glu Ala Met Ile Asp Val Asn Ile Asn Lys
835 840 845
Asn Leu Val Gly Ser Ala Met Ala Gly Ser Ile Gly Gly Tyr Asn Ala
850 855 860
His Ala Ala Asn Ile Val Thr Ala Ile Tyr Ile Ala Cys Gly Gln Met
865 870 875 880
Leu Gly Val Gln Gly Ala Cys Lys Asp Asn Pro Gly Glu Asn Ala Arg
885 890 895
Gln Leu Ala Arg Ile Val Cys Gly Thr Val Met Ala Gly Glu Leu Ser
900 905 910
Leu Met Ala Ala Leu Ala Ala Gly His Leu Val Arg Ser His Met Val
915 920 925
His Asn Arg
930
<210> 38
<211> 2793
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 38
atgacccctg acatgcgcaa gactggccgc tgcgaggcat tcctggagca acgctccatc 60
ttgctcccta atcgcttcct cttttcagga cacccacacg cagcaacctt gacacttcct 120
caggaaggcc caagccaggt ccgctttcgc ggacgaggca cgccgcccgc cggaagcccc 180
atcgcacgcg aaggcggtgt cctcgcaggt gctcctcacg gctctggcac taataaaaaa 240
gtcgtgatgt tggaattgga ccagttgctg gtgggctctg ccgtcgtgcg tgacggtgct 300
gtacctgtgt tgcgtcctgg ctcacgcgac cgcgtcgcca cgatgctgtc tcgcttgttc 360
cgcatgcatg gactttttgt ggcttctcac ccgtgggaag tgatcgtggg tacggtaaca 420
ctcaccatct gcatgatgtc catgaatatg ttcaccggta acaataaaat ttgtggctgg 480
aattatgagt gcccaaagtt cgaagaggac gtcttgtcct ctgatatcat cattttgacc 540
atcacccgtt gcatcgctat tctgtacatc tatttccagt tccaaaacct gcgtcaactg 600
ggttcaaagt acatccttgg tatcgcgggt cttttcacaa ttttttcgtc cttcgtgttc 660
agcactgtgg ttatccactt tttggacaag gaattgacag gacttaatga ggcgctccct 720
ttcttcctcc tgctgattga cctttcccga gcgtcagcat tggcgaaatt tgcattgtcg 780
tctaattctc aagatgaagt ccgagagaac atcgctcgcg gtatggcgat cctgggtccg 840
accttcacgc tcgatgctct ggtagagtgc cttgtcatcg gtgtaggcac catgagcgga 900
gtccgccagt tggagattat gtgttgcttt ggttgtatgt ctgtgctcgc caattacttc 960
gtattcatga cttttttccc tgcctgcgtg tctctcgtcc tcgaactctc ccgagagtcc 1020
cgcgagggcc gtccaatttg gcagctgtca cacttcgcac gagtcctcga ggaagaggaa 1080
aataagccaa acccagttac acagcgtgtt aagatgatca tgagcctggg cctggttctc 1140
gttcacgctc acagccgttg gatcgctgat ccgtctcccc aaaactccac gaccgaacac 1200
tccaaggttt ctcttggttt ggatgaagac gtttcaaagc gcatcgagcc ctccgtttcc 1260
ctctggcagt tttacctgtc caagatgatt tcgatggata tcgagcaggt cgtgaccctt 1320
agcttggcgt tccttttggc agttaagtac atcttcttcg agcaggcaga aacggagtct 1380
acattgtcac tcaagaaccc tatcacgtca ccagtggtga ccccaaaaaa agcacctgat 1440
aattgttgcc gccgtgaacc gctgcttgtt cgccgttccg agaagttgag cagcgtggaa 1500
gaggaaccgg gcgtctccca agaccgcaaa gttgaagtga ttaagccgct cgtcgtcgag 1560
acagaatccg cttcccgcgc tacttttgtc ttgggcgcat ctggaacatc ccctcctgtg 1620
gcggcacgta cacaggagtt ggagattgag ctgccatctg aaccacgtcc caacgaagag 1680
tgcttgcaga tcttggagtc cgccgaaaag ggcgccaagt tcctctctga cgccgaaatc 1740
atccagcttg tgaatgccaa acacattcca gcatacaaat tggaaacgct gatggaaact 1800
catgaacgcg gcgtctctat tcgccgacag cttcttagca ctaaactccc ggagccatcg 1860
tcgttgcagt atctcccata tcgagactac aactactcct tggtgatggg agcatgctgt 1920
gaaaacgtca tcggctacat gccgatccca gtaggcgtgg caggtccact ttgccttgat 1980
ggcaaggagt accaggtccc catggctacc acggagggct gccttgttgc atccacaaac 2040
cgtggctgtc gcgcgatcgg tcttggcgga ggagcatcat cgcgtgtact tgctgatggt 2100
atgacccgcg gccctgtagt tcgtctcccc cgcgcgtgcg atagcgctga ggttaaggcc 2160
tggttggaaa cacctgaggg atttgcagtg atcaaggacg cctttgactc tacctcacgc 2220
ttcgcgcgct tgcagaagct ccacgttacc atggcgggtc gcaacttgta tatccgcttc 2280
cagtccaaaa ccggagacgc tatgggaatg aatatgatct ccaaaggtac cgaaaaagct 2340
ctcctgaagc tccaggaatt cttcccagaa atgcaaattc tcgcggttag cggcaactat 2400
tgcaccgaca agaagccagc agccatcaac tggatcgagg gtcgaggtaa gaccgttgtc 2460
tgcgaggccg ttattccggc taaggtggtc cgtgaggtgc tgaagaccac gacggaagcg 2520
atgatcgatg tgaacatcaa caagaacctc gtcggctccg cgatggcggg ttccattggc 2580
ggttacaatg ctcacgctgc taatatcgta actgctattt atattgcttg cggtcagatg 2640
cttggcgttc aaggtgcatg caaggataac ccaggtgaga acgcccgtca gttggcacgt 2700
atcgtatgcg gcaccgtgat ggcaggcgaa ctttccctta tggccgcact ggcggctggt 2760
cacctcgtcc gttcacacat ggttcataac cgc 2793
<210> 39
<211> 428
<212> PRT
<213> 隐蔽热网菌（Pyrodictium occultum）
<400> 39
Met Gly Glu Glu Arg Ser Leu Arg Asp Arg Leu Glu Glu Val Val Gln
1 5 10 15
Gly Ile Val Glu Gly Arg Ile Arg Leu His Glu Ala Asp Arg Leu Leu
20 25 30
Gly Asn Ala Asn Ala Ala Ala Leu Ala Arg Arg Leu Ala Leu Glu Arg
35 40 45
Met Leu Gly Val Gly Leu Ser Ser Ile Gly Ser Thr Ile Leu Asp Phe
50 55 60
Glu Glu Leu Val Gly Arg Asn Ile Glu Asn Pro Ile Gly Ala Val Gln
65 70 75 80
Ile Pro Leu Gly Val Ala Gly Pro Leu Arg Val His Gly Glu Tyr Ala
85 90 95
Arg Gly Asp Phe Tyr Ile Pro Leu Ala Thr Thr Glu Gly Ala Leu Val
100 105 110
Ala Ser Val Asn Arg Gly Ala Lys Ala Val Thr Leu Ser Gly Gly Ala
115 120 125
Arg Ala Arg Val Leu Arg Asp Gly Met Ala Arg Ala Pro Val Phe Trp
130 135 140
Thr Pro Gly Val Glu Glu Ala Ala Arg Leu Ala Glu Trp Val Gln Glu
145 150 155 160
His Met Glu Glu Val Arg Arg Glu Ala Glu Ser Thr Thr Arg His Gly
165 170 175
Arg Leu Leu Glu Ile Gln Pro Phe Ile Ala Gly Asn Ile Val Trp Leu
180 185 190
Arg Phe Val Tyr Ser Thr Gly Asp Ala Met Gly Met Asn Met Ala Thr
195 200 205
Ile Ala Thr Asp Lys Ala Ala Glu Trp Ile Leu Arg Asn Tyr Pro Gly
210 215 220
Glu Ala Lys Leu Ile Ala Ile Ser Gly Asn Leu Cys Thr Asp Lys Lys
225 230 235 240
Pro Ala Leu Leu Asn Ala Ile Leu Gly Arg Gly Lys Thr Val Val Ala
245 250 255
Glu Ala Thr Ile Lys Arg Asp Ile Ala Leu Arg Val Leu Lys Ala Ala
260 265 270
Pro Glu Asp Ile Asp Thr Val Asn Arg Val Lys Asn Leu Leu Gly Ser
275 280 285
Ala Arg Ala Gly Ser Pro Ser Phe Asn Ala His Tyr Ala Asn Ile Ile
290 295 300
Ala Ala Ile Phe Ile Ala Thr Gly Gln Asp Val Ala Gln Val Val Glu
305 310 315 320
Ser Ser Met Gly Tyr Thr Trp Thr Glu Val Arg Asn Gly Asp Leu Tyr
325 330 335
Ile Ser Val Thr Leu Pro Ser Leu Glu Leu Gly Thr Val Gly Gly Gly
340 345 350
Thr Arg Leu Pro Thr Gln Arg Glu Ala Leu Ala Leu Leu Gly Ala Ala
355 360 365
Gly Gly Gly Asn Pro Pro Gly Ala Asn Ala Lys Lys Leu Ala Glu Val
370 375 380
Thr Ala Ala Ala Val Leu Ala Gly Glu Leu Asn Leu Leu Ala Ala Leu
385 390 395 400
Ala Ala Asn Glu Leu Ala Arg Ala His Arg Leu Leu Gly Arg Gly Glu
405 410 415
Ala Lys Thr Gly Asn Lys Asn Pro Thr Asn His Gly
420 425
<210> 40
<211> 1284
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 40
atgggagaag agcgctctct gcgcgatcgc ctggaagagg tagtccaagg catcgtagag 60
ggtcgcatcc gcctgcatga agcagaccgc ctgctgggaa atgcaaatgc agcagcgttg 120
gctcgccgcc tggcactgga acgcatgctt ggagttggct tgtccagcat tggatcgacg 180
attttggact tcgaagagct tgttggccgt aatattgaaa acccgatcgg tgccgtgcag 240
atccccctcg gtgtcgcggg cccgctgcgc gtccacggcg agtatgctcg cggtgacttc 300
tacattccac ttgctaccac tgaaggcgca cttgtggcat cagtgaatcg cggcgcaaag 360
gctgtgacgc tctctggcgg tgcacgcgcc cgcgtacttc gagatggcat ggcccgagca 420
ccggtgtttt ggactcctgg tgtggaagaa gcagcgcgac tcgcggaatg ggtgcaagag 480
catatggaag aagtccgtcg tgaagcagag tccacgacgc gtcacggacg tctcctcgaa 540
atccagccgt tcatcgctgg caacatcgtt tggcttcgct tcgtctattc taccggcgac 600
gccatgggca tgaacatggc aaccattgca accgataagg cggcagagtg gatcctgcgc 660
aattacccgg gagaagcgaa actcatcgca atctccggca acctctgcac agacaagaag 720
cctgctctct tgaacgctat tctgggccgc ggtaaaaccg tggtcgcgga ggcaactatt 780
aagcgcgaca tcgcattgcg cgtgcttaaa gcagctcccg aggacatcga tactgtgaac 840
cgtgtgaaaa atcttttggg cagcgctcgc gccggttcac cttcgtttaa cgcacactat 900
gctaacatca ttgcagcaat cttcatcgcg acaggacagg atgtggcgca ggtagtggag 960
tccagcatgg gatacacctg gaccgaagtt cgcaatggag acctttacat ttccgtgacg 1020
ctgccctcct tggaactggg caccgttggc ggtggtaccc gcctgcccac acagcgcgag 1080
gcactcgccc tcctgggagc agcaggcggt ggtaacccac cgggtgccaa tgcgaagaag 1140
ttggcagagg ttacagccgc agccgtgctg gccggcgaac tcaatttgtt ggccgctctt 1200
gcagcgaatg aactcgcccg cgcgcaccgt ttgctgggtc gtggcgaagc gaagactggt 1260
aataaaaatc caaccaacca cggt 1284
<210> 41
<211> 352
<212> PRT
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 41
Met Ala Ser Glu Lys Glu Ile Arg Arg Glu Arg Phe Leu Asn Val Phe
1 5 10 15
Pro Lys Leu Val Glu Glu Leu Asn Ala Ser Leu Leu Ala Tyr Gly Met
20 25 30
Pro Lys Glu Ala Cys Asp Trp Tyr Ala His Ser Leu Asn Tyr Asn Thr
35 40 45
Pro Gly Gly Lys Leu Asn Arg Gly Leu Ser Val Val Asp Thr Tyr Ala
50 55 60
Ile Leu Ser Asn Lys Thr Val Glu Gln Leu Gly Gln Glu Glu Tyr Glu
65 70 75 80
Lys Val Ala Ile Leu Gly Trp Cys Ile Glu Leu Leu Gln Ala Tyr Phe
85 90 95
Leu Val Ala Asp Asp Met Met Asp Lys Ser Ile Thr Arg Arg Gly Gln
100 105 110
Pro Cys Trp Tyr Lys Val Pro Glu Val Gly Glu Ile Ala Ile Asn Asp
115 120 125
Ala Phe Met Leu Glu Ala Ala Ile Tyr Lys Leu Leu Lys Ser His Phe
130 135 140
Arg Asn Glu Lys Tyr Tyr Ile Asp Ile Thr Glu Leu Phe His Glu Val
145 150 155 160
Thr Phe Gln Thr Glu Leu Gly Gln Leu Met Asp Leu Ile Thr Ala Pro
165 170 175
Glu Asp Lys Val Asp Leu Ser Lys Phe Ser Leu Lys Lys His Ser Phe
180 185 190
Ile Val Thr Phe Gly Thr Ala Tyr Tyr Ser Phe Tyr Leu Pro Val Ala
195 200 205
Leu Ala Met Tyr Val Ala Gly Ile Thr Asp Glu Lys Asp Leu Lys Gln
210 215 220
Ala Arg Asp Val Leu Ile Pro Leu Gly Glu Tyr Phe Gln Ile Gln Asp
225 230 235 240
Asp Tyr Leu Asp Cys Phe Gly Thr Pro Glu Gln Ile Gly Lys Ile Gly
245 250 255
Thr Asp Ile Gln Asp Asn Lys Cys Ser Trp Val Ile Asn Lys Ala Leu
260 265 270
Glu Leu Ala Ser Ala Glu Gln Arg Lys Thr Leu Asp Glu Asn Tyr Gly
275 280 285
Lys Lys Asp Ser Val Ala Glu Ala Lys Cys Lys Lys Ile Phe Asn Asp
290 295 300
Leu Lys Ile Glu Gln Leu Tyr His Glu Tyr Glu Glu Ser Ile Ala Lys
305 310 315 320
Asp Leu Lys Ala Lys Ile Ser Gln Val Asp Glu Ser Arg Gly Phe Lys
325 330 335
Ala Asp Val Leu Thr Ala Phe Leu Asn Lys Val Tyr Lys Arg Ser Lys
340 345 350
<210> 42
<211> 1056
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 42
atggcgtcgg aaaaagaaat ccgccgcgag cgcttcctga atgtgtttcc aaagctggtc 60
gaggaattga acgcttcgct gcttgcgtat ggcatgccga aagaggcgtg cgattggtac 120
gcacattccc tgaactataa caccccaggc ggtaaactca accgcggtct gtccgttgta 180
gatacatatg ccattcttag caacaagacc gtagaacagt tgggacaaga ggaatatgaa 240
aaggtggcta tcttgggctg gtgtatcgaa ctgttgcaag cctacttcct ggtcgccgat 300
gatatgatgg ataagtccat cacacgacgt ggacagccat gttggtacaa ggtccctgaa 360
gtcggcgaaa tcgctatcaa cgacgcattc atgctcgagg cagcaattta caaacttttg 420
aaaagccatt tccgaaacga gaagtattac atcgacatta ccgagctctt ccatgaagtc 480
accttccaga ccgaactcgg ccaattgatg gatctgatta ccgccccgga ggacaaggtc 540
gatctttcca aattcagcct gaagaaacat tccttcatcg taacctttgg tacagcatac 600
tattcattct atcttcctgt ggcactcgca atgtacgtag ccggtatcac ggatgaaaag 660
gatctgaaac aagcacgcga cgtactcatt cccctgggtg agtactttca gattcaagat 720
gactacctcg attgcttcgg cacgcctgaa caaatcggca aaatcggtac tgacatccag 780
gataataaat gctcctgggt gatcaataag gccctcgagt tggcgtcagc cgagcagcgc 840
aagactctgg acgaaaatta cggtaaaaaa gattcagttg ccgaggcaaa gtgcaaaaag 900
atctttaacg atcttaagat tgaacagctt taccacgaat acgaagaaag cattgctaag 960
gatctgaaag ctaagatttc tcaggtcgat gaatcacgcg gattcaaggc cgatgtgctc 1020
actgctttcc tgaacaaggt ctacaagcgc tcaaaa 1056
<210> 43
<211> 296
<212> PRT
<213> 北美冷杉（Abies grandis）
<400> 43
Met Phe Asp Phe Asn Lys Tyr Met Asp Ser Lys Ala Met Thr Val Asn
1 5 10 15
Glu Ala Leu Asn Lys Ala Ile Pro Leu Arg Tyr Pro Gln Lys Ile Tyr
20 25 30
Glu Ser Met Arg Tyr Ser Leu Leu Ala Gly Gly Lys Arg Val Arg Pro
35 40 45
Val Leu Cys Ile Ala Ala Cys Glu Leu Val Gly Gly Thr Glu Glu Leu
50 55 60
Ala Ile Pro Thr Ala Cys Ala Ile Glu Met Ile His Thr Met Ser Leu
65 70 75 80
Met His Asp Asp Leu Pro Cys Ile Asp Asn Asp Asp Leu Arg Arg Gly
85 90 95
Lys Pro Thr Asn His Lys Ile Phe Gly Glu Asp Thr Ala Val Thr Ala
100 105 110
Gly Asn Ala Leu His Ser Tyr Ala Phe Glu His Ile Ala Val Ser Thr
115 120 125
Ser Lys Thr Val Gly Ala Asp Arg Ile Leu Arg Met Val Ser Glu Leu
130 135 140
Gly Arg Ala Thr Gly Ser Glu Gly Val Met Gly Gly Gln Met Val Asp
145 150 155 160
Ile Ala Ser Glu Gly Asp Pro Ser Ile Asp Leu Gln Thr Leu Glu Trp
165 170 175
Ile His Ile His Lys Thr Ala Met Leu Leu Glu Cys Ser Val Val Cys
180 185 190
Gly Ala Ile Ile Gly Gly Ala Ser Glu Ile Val Ile Glu Arg Ala Arg
195 200 205
Arg Tyr Ala Arg Cys Val Gly Leu Leu Phe Gln Val Val Asp Asp Ile
210 215 220
Leu Asp Val Thr Lys Ser Ser Asp Glu Leu Gly Lys Thr Ala Gly Lys
225 230 235 240
Asp Leu Ile Ser Asp Lys Ala Thr Tyr Pro Lys Leu Met Gly Leu Glu
245 250 255
Lys Ala Lys Glu Phe Ser Asp Glu Leu Leu Asn Arg Ala Lys Gly Glu
260 265 270
Leu Ser Cys Phe Asp Pro Val Lys Ala Ala Pro Leu Leu Gly Leu Ala
275 280 285
Asp Tyr Val Ala Phe Arg Gln Asn
290 295
<210> 44
<211> 888
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 44
atgttcgact tcaacaaata catggacagc aaggcaatga ccgtgaacga ggcgctgaat 60
aaagctattc ctttgcgtta tccgcagaag atctacgaat ccatgcgata ctctttgttg 120
gcaggtggta agcgagtgcg ccccgtgctt tgcattgcgg cttgcgagct ggtcggtggt 180
actgaggaac tggcgatccc aaccgcttgt gctatcgaga tgattcacac gatgtccctt 240
atgcacgatg acctgccctg tatcgacaac gatgatttgc gccgaggcaa gccaacaaat 300
cataagatct tcggcgagga caccgccgtc actgccggca atgctctgca ctcgtatgcc 360
ttcgagcaca ttgccgtgtc tacttccaaa acggtgggtg cagatcgcat tctgcgcatg 420
gtatccgagt tgggtcgcgc aactggctcg gagggcgtca tgggtggtca aatggtcgac 480
atcgcatctg aaggcgatcc atcgatcgac ctgcagacat tggaatggat ccacatccac 540
aagactgcga tgcttttgga atgttccgtg gtgtgcggag ctattattgg cggagcatct 600
gagatcgtga tcgagcgcgc tcgccgttat gctcgttgcg tgggtttgct gtttcaggtc 660
gtggacgaca ttctcgatgt caccaaatcc tccgatgagt tgggtaaaac tgccggcaaa 720
gacttgattt ctgacaaggc gacctacccc aagcttatgg gtctcgaaaa agccaaggag 780
ttctcagatg aactcctcaa ccgcgcaaag ggagaactct catgctttga tccagtgaag 840
gcagccccat tgcttggttt ggctgattac gtggccttcc gccagaat 888
<210> 45
<211> 297
<212> PRT
<213> 挪威云杉（Picea abies）
<400> 45
Met Glu Phe Asp Phe Asp Lys Tyr Met His Ser Lys Ala Ile Ala Val
1 5 10 15
Asn Glu Ala Leu Asp Lys Val Ile Pro Pro Arg Tyr Pro Gln Lys Ile
20 25 30
Tyr Glu Ser Met Arg Tyr Ser Leu Leu Ala Gly Gly Lys Arg Val Arg
35 40 45
Pro Ile Leu Cys Ile Ala Ala Cys Glu Leu Met Gly Gly Thr Glu Glu
50 55 60
Leu Ala Met Pro Thr Ala Cys Ala Ile Glu Met Ile His Thr Met Ser
65 70 75 80
Leu Ile His Asp Asp Leu Pro Tyr Ile Asp Asn Asp Asp Leu Arg Arg
85 90 95
Gly Lys Pro Thr Asn His Lys Val Phe Gly Glu Asp Thr Ala Ile Ile
100 105 110
Ala Gly Asp Ala Leu Leu Ser Leu Ala Phe Glu His Val Ala Val Ser
115 120 125
Thr Ser Arg Thr Leu Gly Thr Asp Ile Ile Leu Arg Leu Leu Ser Glu
130 135 140
Ile Gly Arg Ala Thr Gly Ser Glu Gly Val Met Gly Gly Gln Val Val
145 150 155 160
Asp Ile Glu Ser Glu Gly Asp Pro Ser Ile Asp Leu Glu Thr Leu Glu
165 170 175
Trp Val His Ile His Lys Thr Ala Val Leu Leu Glu Cys Ser Val Val
180 185 190
Cys Gly Ala Ile Met Gly Gly Ala Ser Glu Asp Asp Ile Glu Arg Ala
195 200 205
Arg Arg Tyr Ala Arg Cys Val Gly Leu Leu Phe Gln Val Val Asp Asp
210 215 220
Ile Leu Asp Val Ser Gln Ser Ser Glu Glu Leu Gly Lys Thr Ala Gly
225 230 235 240
Lys Asp Leu Ile Ser Asp Lys Ala Thr Tyr Pro Lys Leu Met Gly Leu
245 250 255
Glu Lys Ala Lys Glu Phe Ala Asp Glu Leu Leu Asn Arg Gly Lys Gln
260 265 270
Glu Leu Ser Cys Phe Asp Pro Thr Lys Ala Ala Pro Leu Phe Ala Leu
275 280 285
Ala Asp Tyr Ile Ala Ser Arg Gln Asn
290 295
<210> 46
<211> 891
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 46
atggaatttg atttcgataa gtacatgcac agcaaagcaa ttgccgtcaa cgaggctctc 60
gataaagtta tcccacctcg ttacccacag aaaatctacg aaagcatgcg ctattccctg 120
ctggcaggcg gcaaacgcgt acgcccaatc ctctgcatcg cggcctgcga acttatgggc 180
ggcaccgagg agctcgctat gccaaccgca tgcgcgattg agatgatcca caccatgagc 240
cttattcacg atgatctgcc atatatcgac aacgacgatt tgcgccgtgg aaagcctacc 300
aaccacaaag tcttcggaga ggacaccgcc atcattgctg gcgacgctct gctgtccctg 360
gcttttgagc acgttgcagt cagcacctcc cgcacgctcg gcacggatat tatcctgcgc 420
ctcctcagcg aaatcggtcg cgctaccggt tccgaaggtg tgatgggcgg tcaggtggtg 480
gatatcgaat ctgaaggcga tcccagcatc gaccttgaaa cgctggaatg ggtccatatc 540
cataagaccg cggttctgct ggaatgctct gttgtttgcg gtgcaatcat gggcggcgca 600
tcagaggatg acatcgagcg cgcacgtcgc tacgctcgtt gcgtaggctt gttgtttcaa 660
gtcgtggacg atatcctcga tgtatcccag tcttccgaag agctgggtaa gactgcgggc 720
aaggatctga tcagcgataa agccacctac ccaaaactta tgggtcttga gaaagcgaag 780
gaattcgctg acgaactgct gaaccgcggc aagcaggagc tgtcctgctt tgatccaacc 840
aaagccgcac cgctcttcgc tctggcagat tacattgcga gccgtcaaaa c 891
<210> 47
<211> 355
<212> PRT
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 47
Met Thr Pro Asp Val Thr Asn Asp Ala Ser Gln Glu Thr Val Asp Leu
1 5 10 15
Val Arg Arg Thr Ser Ala Asp Leu Leu Arg Arg Val Glu Ala Arg Ile
20 25 30
Gln Ser Leu Leu Ser Ala Gly His Asp Thr Trp Ala Ala Val Asp Glu
35 40 45
Arg Ala Val Val Pro Ile Asp Ala Leu Ala Glu Leu Met Ala Ser Gly
50 55 60
Gly Lys Arg Ile Arg Pro Ala Phe Cys Ile Ala Gly His Leu Ala Ala
65 70 75 80
Gly Gly Asp Pro Asp Asp Ser Gly Val Val Ala Ala Gly Ala Ala Leu
85 90 95
Glu Met Leu His Ala Ser Leu Leu Val His Asp Asp Ile Leu Asp Asp
100 105 110
Ser Ser Gln Arg Arg Gly Leu Pro Thr Val His Ala Lys His Thr Ala
115 120 125
Leu His Gln Asp Leu Gly Trp Gln Gly Glu Ser Arg Arg Phe Gly Glu
130 135 140
Gly Val Gly Leu Leu Val Gly Gly Leu Ala Leu Thr Tyr Ala Asp Glu
145 150 155 160
Leu Met Cys Glu Ala Pro Ala Thr Ala Ile Ala Glu Trp Asn Lys Leu
165 170 175
Arg Ser Glu Val Leu Ile Gly Gln Tyr Met Asp Val Val Ala Ala Ala
180 185 190
Glu Phe Ser Val Asp Pro Arg Leu Ser Arg Leu Ile Ala Val Ile Lys
195 200 205
Ser Gly Arg Tyr Thr Ile His Arg Pro Leu Val Ile Gly Ala Asn Val
210 215 220
Ala Gly Arg Thr Asp Leu Asp Ala Ala Phe Ala Glu Tyr Gly Glu Ala
225 230 235 240
Val Gly Glu Ala Phe Gln Leu Arg Asp Asp Leu Leu Asp Ala Phe Gly
245 250 255
Asp Ser Ala Ala Thr Gly Lys Pro Ala Gly Leu Asp Phe Thr Gln His
260 265 270
Lys Met Thr Leu Leu Leu Gly Trp Ala Met Gln Arg Asp Glu Ser Ile
275 280 285
Arg Ala Leu Ile Thr Glu Pro Gly His Ser Ala Asp Asp Val Arg Arg
290 295 300
His Leu Leu Asp Thr Lys Val Pro Asp Asp Val Glu Gln His Ile Ala
305 310 315 320
Gly Leu Val Glu Arg Gly Cys Lys Ala Ile Thr Asp Ala Pro Ile Ala
325 330 335
Pro Val Trp Arg Glu Glu Leu Met Glu Met Ala Asn Arg Val Ala Tyr
340 345 350
Arg Asp Ala
355
<210> 48
<211> 1065
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 48
atgaccccag acgtgaccaa cgatgcctca caagaaaccg tcgatctggt tcgccgaacc 60
tctgcggatc tcctccgccg cgttgaagct cgcattcagt cccttctctc cgcgggccac 120
gatacctggg ctgccgtcga tgagcgagcc gttgttccaa ttgatgccct cgcagagctc 180
atggcttccg gtggaaaacg catccgcccg gcattctgca ttgccggcca cttggccgcc 240
ggtggagatc cagacgattc cggcgtcgtc gcggctggcg ccgcgctcga gatgcttcac 300
gcatcccttc tggtgcacga cgatatcctg gatgattcct ctcagcgccg cggcctccca 360
accgtccacg caaagcacac ggccctgcat caggatttgg gatggcaagg agagtcacgt 420
cgctttggag agggtgtggg actcttggtt ggtggccttg cacttaccta cgccgacgaa 480
ttgatgtgcg aagcgccagc aacagccatt gccgagtgga acaagctgcg atccgaagtg 540
cttatcggcc agtacatgga tgtggtcgca gctgcggagt ttagcgtgga cccacgcctt 600
tctcgcttga tcgctgtcat taaatcgggc cgctacacta ttcaccgtcc actggtcatt 660
ggtgctaacg tcgcgggccg caccgacttg gacgctgcgt tcgctgaata cggtgaagct 720
gtgggcgagg ctttccaact gcgcgatgac ctcctcgatg catttggtga tagcgccgca 780
accggtaagc cagcaggtct tgactttacc caacataaga tgaccctctt gttgggctgg 840
gcaatgcaac gcgacgagtc cattcgagcc ctcattaccg agcccggcca ctcggccgat 900
gacgttcgtc gccacctgct tgacaccaag gtgccagacg atgttgagca gcacattgcc 960
ggtttggtgg agcgaggatg caaagctatc acagacgcac caattgcgcc cgtgtggcgc 1020
gaagaactta tggaaatggc gaatcgcgtt gcatatcgcg atgct 1065
<210> 49
<211> 415
<212> PRT
<213> 番茄（Solanum lycopersicum）
<400> 49
Met Ile Phe Ser Lys Gly Leu Ala Gln Ile Ser Arg Asn Arg Phe Ser
1 5 10 15
Arg Cys Arg Trp Leu Phe Ser Leu Arg Pro Ile Pro Gln Leu His Gln
20 25 30
Ser Asn His Ile His Asp Pro Pro Lys Val Leu Gly Cys Arg Val Ile
35 40 45
His Ser Trp Val Ser Asn Ala Leu Ser Gly Ile Gly Gln Gln Ile His
50 55 60
Gln Gln Ser Thr Ala Val Ala Glu Glu Gln Val Asp Pro Phe Ser Leu
65 70 75 80
Val Ala Asp Glu Leu Ser Leu Leu Thr Asn Arg Leu Arg Ser Met Val
85 90 95
Val Ala Glu Val Pro Lys Leu Ala Ser Ala Ala Glu Tyr Phe Phe Lys
100 105 110
Leu Gly Val Glu Gly Lys Arg Phe Arg Pro Thr Val Leu Leu Leu Met
115 120 125
Ala Thr Ala Leu Asn Val Gln Ile Pro Arg Ser Ala Pro Gln Val Asp
130 135 140
Val Asp Ser Phe Ser Gly Asp Leu Arg Thr Arg Gln Gln Cys Ile Ala
145 150 155 160
Glu Ile Thr Glu Met Ile His Val Ala Ser Leu Leu His Asp Asp Val
165 170 175
Leu Asp Asp Ala Asp Thr Arg Arg Gly Ile Gly Ser Leu Asn Phe Val
180 185 190
Met Gly Asn Lys Leu Ala Val Leu Ala Gly Asp Phe Leu Leu Ser Arg
195 200 205
Ala Cys Val Ala Leu Ala Ser Leu Lys Asn Thr Glu Val Val Cys Leu
210 215 220
Leu Ala Thr Val Val Glu His Leu Val Thr Gly Glu Thr Met Gln Met
225 230 235 240
Thr Thr Ser Ser Asp Glu Arg Cys Ser Met Glu Tyr Tyr Met Gln Lys
245 250 255
Thr Tyr Tyr Lys Thr Ala Ser Leu Ile Ser Asn Ser Cys Lys Ala Ile
260 265 270
Ala Leu Leu Ala Gly His Ser Ala Glu Val Ser Val Leu Ala Phe Asp
275 280 285
Tyr Gly Lys Asn Leu Gly Leu Ala Phe Gln Leu Ile Asp Asp Val Leu
290 295 300
Asp Phe Thr Gly Thr Ser Ala Thr Leu Gly Lys Gly Ser Leu Ser Asp
305 310 315 320
Ile Arg His Gly Ile Val Thr Ala Pro Ile Leu Tyr Ala Met Glu Glu
325 330 335
Phe Pro Gln Leu Arg Thr Leu Val Asp Arg Gly Phe Asp Asp Pro Val
340 345 350
Asn Val Glu Ile Ala Leu Asp Tyr Leu Gly Lys Ser Arg Gly Ile Gln
355 360 365
Arg Thr Arg Glu Leu Ala Arg Lys His Ala Ser Leu Ala Ser Ala Ala
370 375 380
Ile Asp Ser Leu Pro Glu Ser Asp Asp Glu Glu Val Gln Arg Ser Arg
385 390 395 400
Arg Ala Leu Val Glu Leu Thr His Arg Val Ile Thr Arg Thr Lys
405 410 415
<210> 50
<211> 1245
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 50
atgattttca gcaaaggatt ggcgcaaatt tctcgcaacc gtttttcacg ctgccgttgg 60
ctgttttcgc tgcgaccaat ccctcagctc caccagagca atcatattca tgatccaccc 120
aaggttcttg gttgccgagt aatccacagc tgggtttcca atgcgctgtc cggtatcggc 180
cagcagatcc accagcagtc taccgcagtg gcggaggaac aagtggatcc tttctccctc 240
gtggctgatg aactctccct cctgaccaat cgattgcgct ctatggtcgt cgccgaagtc 300
cccaaattgg cttccgcggc ggagtacttt ttcaagttgg gcgtggaggg aaaacgcttc 360
cgaccaaccg tcttgttgct catggctact gcgctgaatg tccagatccc ccgctccgca 420
ccgcaagttg atgtggatag cttctcaggc gatcttcgta cccgacagca gtgtatcgcc 480
gagattaccg agatgatcca cgttgcttca ttgctccatg atgacgtgct tgatgatgct 540
gatacccgtc gaggcatcgg ctcactgaat ttcgttatgg gcaataagct tgctgttctc 600
gctggtgatt ttttgctgtc ccgcgcgtgc gtggcattgg cctctctgaa aaacaccgag 660
gtggtttgcc ttcttgcaac agttgttgag catcttgtaa ccggtgagac catgcagatg 720
accacctcgt ccgatgaacg atgcagcatg gagtactaca tgcaaaagac ctactataaa 780
acggcgtccc tgatctcgaa ctcttgtaaa gctattgcgc tcctggctgg ccactctgcg 840
gaagtttcgg tgttggcatt tgattacggt aaaaatctcg gcttggcttt ccaattgatc 900
gatgatgttc ttgatttcac tggaacttcc gcaaccttgg gcaaaggttc cctgtccgat 960
attcgccacg gcatcgtcac cgccccaatc ttgtatgcga tggaagagtt ccctcagctc 1020
cgtaccctcg ttgatcgtgg cttcgatgat ccggtaaacg tggaaattgc cctcgactac 1080
ttgggtaagt ctcgcggcat ccaacgcaca cgcgagcttg cacgtaaaca cgcgtctctc 1140
gcgagcgcgg cgatcgactc cctgccggaa tccgatgatg aagaggtgca gcgcagccgc 1200
cgtgcgcttg ttgaattgac tcaccgcgtt atcacccgta ctaag 1245
<210> 51
<211> 420
<212> PRT
<213> 长春花（Catharanthus roseus）
<400> 51
Met Leu Phe Ser Arg Gly Leu Tyr Arg Ile Ala Arg Thr Ser Leu Asn
1 5 10 15
Arg Ser Arg Leu Leu Tyr Pro Leu Gln Ser Gln Ser Pro Glu Leu Leu
20 25 30
Gln Ser Phe Gln Phe Arg Ser Pro Ile Gly Ser Ser Gln Lys Val Ser
35 40 45
Gly Phe Arg Val Ile Tyr Ser Trp Val Ser Ser Ala Leu Ala Asn Val
50 55 60
Gly Gln Gln Val Gln Arg Gln Ser Asn Ser Val Ala Glu Glu Pro Leu
65 70 75 80
Asp Pro Phe Ser Leu Val Ala Asp Glu Leu Ser Ile Leu Ala Asn Arg
85 90 95
Leu Arg Ser Met Val Val Ala Glu Val Pro Lys Leu Ala Ser Ala Ala
100 105 110
Glu Tyr Phe Phe Lys Leu Gly Val Glu Gly Lys Arg Phe Arg Pro Thr
115 120 125
Val Leu Leu Leu Met Ala Thr Ala Ile Asp Ala Pro Ile Ser Arg Thr
130 135 140
Pro Pro Asp Thr Ser Leu Asp Thr Leu Ser Thr Glu Leu Arg Leu Arg
145 150 155 160
Gln Gln Thr Ile Ala Glu Ile Thr Lys Met Ile His Val Ala Ser Leu
165 170 175
Leu His Asp Asp Val Leu Asp Asp Ala Glu Thr Arg Arg Gly Ile Gly
180 185 190
Ser Leu Asn Phe Val Met Gly Asn Lys Leu Ala Val Leu Ala Gly Asp
195 200 205
Phe Leu Leu Ser Arg Ala Cys Val Ala Leu Ala Ser Leu Lys Asn Thr
210 215 220
Glu Val Val Ser Leu Leu Ala Thr Val Val Glu His Leu Val Thr Gly
225 230 235 240
Glu Thr Met Gln Met Thr Thr Thr Ser Asp Gln Arg Cys Ser Met Glu
245 250 255
Tyr Tyr Met Gln Lys Thr Tyr Tyr Met Thr Ala Ser Leu Ile Ser Asn
260 265 270
Ser Cys Lys Ala Ile Ala Leu Leu Ala Gly Gln Thr Ser Glu Val Ala
275 280 285
Met Leu Ala Tyr Glu Tyr Gly Lys Asn Leu Gly Leu Ala Phe Gln Leu
290 295 300
Ile Asp Asp Val Leu Asp Phe Thr Gly Thr Ser Ala Ser Leu Gly Lys
305 310 315 320
Gly Ser Leu Ser Asp Ile Arg His Gly Ile Val Thr Ala Pro Ile Leu
325 330 335
Phe Ala Ile Glu Glu Phe Pro Glu Leu Arg Ala Val Val Asp Glu Gly
340 345 350
Phe Glu Asn Pro Tyr Asn Val Asp Leu Ala Leu His Tyr Leu Gly Lys
355 360 365
Ser Arg Gly Ile Gln Arg Thr Arg Glu Leu Ala Ile Lys His Ala Asn
370 375 380
Leu Ala Ser Asp Ala Ile Asp Ser Leu Pro Val Thr Asp Asp Glu His
385 390 395 400
Val Leu Arg Ser Arg Arg Ala Leu Val Glu Leu Thr Gln Arg Val Ile
405 410 415
Thr Arg Arg Lys
420
<210> 52
<211> 1260
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 52
atgctgtttt cacgcggcct ctatcgcatc gcgcgaacct ccctgaaccg tagccgcctc 60
ttgtacccat tgcaatcaca aagccccgag ttgctccagt cgtttcagtt ccgctcgcct 120
attggatcat cccaaaaggt atccggattc cgtgttatct actcctgggt gtcctcggca 180
ttggccaacg tcggccagca ggtacaacga caaagcaact ccgtagcgga ggagcccctg 240
gatccatttt cactggttgc cgatgaactt tccattctcg ctaaccgtct ccgctcaatg 300
gtcgtcgctg aagtgcctaa gctcgcttcc gcagcagagt acttcttcaa gttgggtgtg 360
gagggcaagc gtttccgacc caccgtcctt ctgttgatgg cgaccgctat cgacgcaccc 420
atctctcgca caccacctga tacctcgttg gatacccttt ctactgagct tcgcctgcgc 480
caacagacca ttgccgagat caccaaaatg attcacgttg cttcgctcct ccacgatgat 540
gtgctcgatg acgctgagac ccgccgagga attggttcac ttaattttgt aatgggtaac 600
aagcttgctg tgctcgcagg tgatttcctg ctctcacgcg catgtgtggc cctggcttca 660
ctgaaaaata ccgaggtggt ttcgttgctg gccactgttg tagagcacct cgtcactggc 720
gagactatgc aaatgacaac cacgtcggac cagcgctgca gcatggagta ttatatgcag 780
aaaacttatt acatgaccgc gtctctgatt tcgaactcct gcaaggccat tgcgttgctc 840
gccggacaaa cctcggaggt cgccatgctt gcatatgaat acggtaaaaa cctgggattg 900
gccttccagc tcatcgacga tgtgctggat ttcactggaa caagcgctag cctgggtaag 960
ggttccttga gcgatatccg ccatggcatc gtaacggccc caattctctt cgcgattgaa 1020
gagttccctg agttgcgtgc tgtagttgat gaaggctttg agaacccata taacgtagac 1080
cttgctttgc attacctcgg taaatctcgt ggtattcaac gtacccgcga actggcgatt 1140
aaacacgcca atcttgcatc agatgcgatc gattctttgc cggttaccga tgacgaacac 1200
gtgctgcgct ctcgtcgcgc acttgtggaa cttacccagc gtgtgatcac gcgtcgcaaa 1260
<210> 53
<211> 367
<212> PRT
<213> 谷氨酸棒状杆菌（C. glutamicum）
<400> 53
Met Ser Thr Phe Asp Ala His Asp Leu Asp Leu Asp Thr Phe Pro Glu
1 5 10 15
Val Val Arg Asp Arg Leu Thr Gln Phe Leu Asp Ala Gln Ala Thr Thr
20 25 30
Ile Ala Gly Ile Gly Asp Pro Val Thr Glu Ala Val Ser His Leu Arg
35 40 45
Ser Phe Val Leu Asn Gly Gly Lys Arg Ile Arg Pro Leu Tyr Ala Trp
50 55 60
Ala Gly Phe Leu Ala Ala Gln Gly Leu Glu Asn Ser Gly Glu Lys Ile
65 70 75 80
Glu Ala Val Leu Asp Ala Ala Ser Ser Leu Glu Phe Ile Gln Ala Tyr
85 90 95
Phe Leu Ile His Asp Asp Pro Ala Ile Asp Ser Ser Asp Thr Arg Arg
100 105 110
Gly Ala Pro Thr Val His Arg Ala Val Glu Ala Asn His Arg Ala His
115 120 125
Asn Leu Glu Gly Asp Ser Glu His Phe Gly Glu Ser Val Ser Ile Leu
130 135 140
Ala Gly Asp Met Ala Leu Val Trp Ala Glu Asp Met Leu Gln Asp Ser
145 150 155 160
Gly Leu Ser Ala Glu Ala Leu Ala Arg Thr Arg Asp Ala Trp Arg Gly
165 170 175
Met Arg Thr Glu Val Ile Gly Gly Gln Leu Leu Asp Ile Tyr Leu Glu
180 185 190
Ala His Ala Asn Glu Ala Val Glu Leu Ala Asp Ser Val Asn Arg Phe
195 200 205
Lys Thr Ala Ala Tyr Thr Ile Ala Arg Pro Leu His Leu Gly Ala Ser
210 215 220
Ile Ala Gly Gly Ser Gln Glu Leu Ile Asp Ala Leu Leu His Tyr Gly
225 230 235 240
His Asp Ile Gly Ile Ala Phe Gln Leu Arg Asp Asp Leu Leu Gly Val
245 250 255
Phe Gly Asp Pro Lys Val Thr Gly Lys Pro Ala Gly Asp Asp Ile Arg
260 265 270
Glu Gly Lys Arg Thr Val Leu Leu Ala Leu Ala Leu Gln Arg Ala Asp
275 280 285
Lys Asn Ser Pro Glu Ala Ala Ala Ala Ile Arg Ala Gly Ile Gly Lys
290 295 300
Val Ser Thr Pro Glu Asp Ile Ala Val Ile Ala Lys His Ile Arg Ala
305 310 315 320
Thr Gly Ala Glu Glu Glu Val Glu Gln Arg Ile Thr Gln Leu Thr Glu
325 330 335
Ser Gly Leu Ala His Leu Asp Asp Val Glu Ile Pro Asp Glu Val Arg
340 345 350
Thr Gln Leu Arg Ala Leu Ala Ile Arg Ser Thr Glu Arg Arg Met
355 360 365
<210> 54
<211> 1101
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 54
atgtctacat tcgatgctca tgacttggac ctggatacct tccctgaagt tgtgcgcgac 60
cgcctgacgc agtttctcga tgcacaggca acgacgattg cgggtatcgg agaccccgtt 120
accgaggccg tatctcactt gcgttcgttt gtcctcaacg gaggtaagcg tatccgcccc 180
ctttatgcct gggccggctt tttggccgca caaggcctgg agaatagcgg cgaaaagatt 240
gaggcggtgt tggatgcagc gagcagcttg gaattcatcc aagcttattt tcttattcac 300
gatgacccag caattgattc ctccgacacc cgtcgcggcg cacctaccgt ccaccgcgct 360
gtggaagcca accaccgcgc gcacaacttg gagggagaca gcgaacactt cggtgagtcg 420
gtttccatcc ttgcaggtga tatggccctg gtttgggcag aggacatgct tcaggattct 480
ggcctttccg cagaagcgtt ggctcgtacc cgtgacgcct ggcgcggcat gcgcaccgag 540
gtcatcggtg gccagctgct ggacatctat ctggaagctc atgctaatga agccgtcgaa 600
ctggcagaca gcgtcaaccg ttttaagact gccgcatata ctattgcccg ccctcttcat 660
cttggtgcct caatcgctgg cggctcccag gaattgatcg atgcactgct ccactacgga 720
cacgatatcg gtatcgcatt ccagctccga gatgatctgc ttggtgtttt tggtgacccg 780
aaggtaacag gaaaaccagc cggcgatgac atccgtgaag gcaaacgcac agtgcttctc 840
gcattggcgt tgcagcgcgc agacaagaac agccctgagg cagctgccgc aatccgcgcg 900
ggcatcggca aggtgtccac cccggaagat attgctgtca tcgctaaaca tattcgtgcg 960
acaggcgcgg aagaagaagt agagcaacgc atcactcaac tgactgaatc aggcttggcg 1020
catttggatg atgtggaaat tcctgacgag gtgcgtaccc aactgcgcgc gctcgcaatc 1080
cgctccacgg agcgccgaat g 1101
<210> 55
<211> 296
<212> PRT
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 55
Met Thr Asn Lys Leu Thr Ser Phe Leu Ala Asp Arg Lys Lys Thr Ile
1 5 10 15
Glu Asn Gln Leu Ser Val Tyr Thr Glu Lys Leu Asp Met Pro Asp Ser
20 25 30
Leu Lys Lys Ser Met Leu Tyr Ser Leu Gln Ala Gly Gly Lys Arg Leu
35 40 45
Arg Pro Leu Ile Val Leu Ala Val Leu Asn Ala Tyr Gly Lys Ser Glu
50 55 60
Lys Asp Gly Ile Pro Val Gly Cys Ala Val Glu Met Ile His Thr Tyr
65 70 75 80
Phe Leu Ile His Asp Asp Leu Pro Cys Met Asp Asp Asp Asp Leu Arg
85 90 95
Arg Gly Lys Pro Thr Asn His Lys Val Phe Gly Glu Ala Thr Ala Val
100 105 110
Leu Ala Gly Asp Gly Leu Leu Thr Glu Ser Phe Lys Leu Ile Thr Ser
115 120 125
His Val Ser Asp Glu Val Ser Ala Glu Lys Arg Leu Arg Leu Val Asn
130 135 140
Glu Leu Ile Ser Ala Ala Gly Thr Glu Gly Met Val Gly Gly Gln Val
145 150 155 160
Ala Asp Met Glu Ala Glu Asn Arg Gln Val Thr Leu Glu Glu Leu Glu
165 170 175
Ser Ile His Glu Arg Lys Thr Ala Lys Leu Leu Gly Phe Cys Val Ile
180 185 190
Ala Gly Ala Ile Leu Ala Asp Ala Pro Glu Glu Asp Ile Glu Thr Leu
195 200 205
Arg Thr Phe Ser Ser His Ile Gly Ile Gly Phe Gln Ile Arg Asp Asp
210 215 220
Ile Leu Asp Leu Glu Gly Ser Glu Glu Lys Ile Gly Lys Arg Val Gly
225 230 235 240
Ser Asp Thr Thr Asn Asp Lys Ser Thr Tyr Pro Ser Leu Leu Ser Leu
245 250 255
Glu Gly Ala Lys His Lys Leu Asp Val His Ile Lys Glu Ala Lys Arg
260 265 270
Leu Ile Gly Gly Leu Ser Leu Gln Lys Asp Leu Leu Tyr Glu Leu Cys
275 280 285
Asp Leu Ile Ala Ala Arg Asp His
290 295
<210> 56
<211> 888
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 56
atgactaata agctgacgtc cttcctcgca gaccgtaaaa agactatcga aaatcaattg 60
tctgtgtaca cagaaaagct cgatatgcct gattccctca agaagtctat gctttactcc 120
cttcaggcag gtggcaagcg tttgcgcccc ctcatcgtgc ttgcggttct caacgcctac 180
ggtaagtcag agaaagatgg tatcccggtt ggatgcgccg tggaaatgat ccacacctat 240
tttcttatcc acgacgatct tccgtgtatg gacgatgacg accttcgccg tggtaagcca 300
actaaccaca aggtttttgg tgaggcaact gcggttcttg caggtgatgg tctgcttacc 360
gaatccttta aacttatcac ttcccatgtt tctgatgaag tgtctgcaga aaaacgtctg 420
cgcctggtga acgaactcat cagcgccgcg ggcaccgaag gtatggtagg aggacaagtg 480
gctgatatgg aagccgagaa ccgtcaggtt acgcttgaag aactcgagtc tattcacgaa 540
cgcaagaccg ccaagctctt gggcttctgt gttattgctg gagccatcct tgcagatgca 600
cctgaagagg atattgaaac attgcgtacc ttttcatccc acatcggcat cggttttcag 660
atccgagacg acattcttga tctcgagggc tccgaagaaa agattggtaa acgcgtgggc 720
tccgacacca ccaatgacaa gtctacttac ccgtctcttc ttagccttga aggcgcaaag 780
cataagttgg acgttcatat taaggaggcg aagcgtttga ttggtggtct gagcctccag 840
aaagacttgc tctacgagct gtgcgatctc atcgcagcac gcgatcac 888
<210> 57
<211> 396
<212> PRT
<213> 酿酒酵母（S. cerevisiae）S288c
<400> 57
Met Thr Val Tyr Thr Ala Ser Val Thr Ala Pro Val Asn Ile Ala Thr
1 5 10 15
Leu Lys Tyr Trp Gly Lys Arg Asp Thr Lys Leu Asn Leu Pro Thr Asn
20 25 30
Ser Ser Ile Ser Val Thr Leu Ser Gln Asp Asp Leu Arg Thr Leu Thr
35 40 45
Ser Ala Ala Thr Ala Pro Glu Phe Glu Arg Asp Thr Leu Trp Leu Asn
50 55 60
Gly Glu Pro His Ser Ile Asp Asn Glu Arg Thr Gln Asn Cys Leu Arg
65 70 75 80
Asp Leu Arg Gln Leu Arg Lys Glu Met Glu Ser Lys Asp Ala Ser Leu
85 90 95
Pro Thr Leu Ser Gln Trp Lys Leu His Ile Val Ser Glu Asn Asn Phe
100 105 110
Pro Thr Ala Ala Gly Leu Ala Ser Ser Ala Ala Gly Phe Ala Ala Leu
115 120 125
Val Ser Ala Ile Ala Lys Leu Tyr Gln Leu Pro Gln Ser Thr Ser Glu
130 135 140
Ile Ser Arg Ile Ala Arg Lys Gly Ser Gly Ser Ala Cys Arg Ser Leu
145 150 155 160
Phe Gly Gly Tyr Val Ala Trp Glu Met Gly Lys Ala Glu Asp Gly His
165 170 175
Asp Ser Met Ala Val Gln Ile Ala Asp Ser Ser Asp Trp Pro Gln Met
180 185 190
Lys Ala Cys Val Leu Val Val Ser Asp Ile Lys Lys Asp Val Ser Ser
195 200 205
Thr Gln Gly Met Gln Leu Thr Val Ala Thr Ser Glu Leu Phe Lys Glu
210 215 220
Arg Ile Glu His Val Val Pro Lys Arg Phe Glu Val Met Arg Lys Ala
225 230 235 240
Ile Val Glu Lys Asp Phe Ala Thr Phe Ala Lys Glu Thr Met Met Asp
245 250 255
Ser Asn Ser Phe His Ala Thr Cys Leu Asp Ser Phe Pro Pro Ile Phe
260 265 270
Tyr Met Asn Asp Thr Ser Lys Arg Ile Ile Ser Trp Cys His Thr Ile
275 280 285
Asn Gln Phe Tyr Gly Glu Thr Ile Val Ala Tyr Thr Phe Asp Ala Gly
290 295 300
Pro Asn Ala Val Leu Tyr Tyr Leu Ala Glu Asn Glu Ser Lys Leu Phe
305 310 315 320
Ala Phe Ile Tyr Lys Leu Phe Gly Ser Val Pro Gly Trp Asp Lys Lys
325 330 335
Phe Thr Thr Glu Gln Leu Glu Ala Phe Asn His Gln Phe Glu Ser Ser
340 345 350
Asn Phe Thr Ala Arg Glu Leu Asp Leu Glu Leu Gln Lys Asp Val Ala
355 360 365
Arg Val Ile Leu Thr Gln Val Gly Ser Gly Pro Gln Glu Thr Asn Glu
370 375 380
Ser Leu Ile Asp Ala Lys Thr Gly Leu Pro Lys Glu
385 390 395
<210> 58
<211> 1188
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 58
atgaccgttt acaccgcctc agtcaccgca ccagttaaca ttgcaaccct taaatactgg 60
ggcaagcgcg atactaagct taaccttcct accaactcat cgatctcggt gaccctgtct 120
caggatgatc tgcgaacgct tacttctgcg gcaacagcac cagaattcga gcgagatacg 180
ctctggctga atggcgaacc gcattccatt gacaacgagc gcacccagaa ctgcttgcgt 240
gatcttcgcc aactccgtaa ggagatggaa agcaaagatg caagcctccc aaccctctct 300
caatggaaac ttcacatcgt ctctgaaaac aacttcccta cggcggctgg cctggcgtcc 360
tcggcagctg gcttcgctgc attggtttcc gccattgcaa agctttatca gcttccgcaa 420
tcgacctccg agattagccg catcgcccgc aagggttcag gttcggcatg tcgctctctc 480
tttggaggct acgtggcctg ggaaatggga aaggccgaag atggtcacga ttccatggcg 540
gtgcaaatcg ccgatagctc agattggcct caaatgaagg cctgtgtgtt ggtggtatcc 600
gacatcaaaa aagacgtttc ctcgactcaa ggaatgcaac ttacggtagc tacgagcgaa 660
ctgtttaagg agcgcattga gcatgtggtg cccaaacgct ttgaagtcat gcgtaaggca 720
atcgtggaga aggacttcgc gaccttcgcg aaggaaacta tgatggatag caacagcttc 780
cacgctacgt gtctggattc gttccctccg attttctata tgaatgacac gtctaaacgc 840
atcatttcct ggtgtcacac gatcaaccaa ttctacggcg aaactatcgt agcatacact 900
tttgacgcag gaccgaacgc cgttctctac tacctcgccg aaaacgagtc aaaattgttc 960
gccttcatct acaaactctt tggatccgtc cctggttggg acaagaagtt cacgactgaa 1020
cagttggaag cattcaacca tcagtttgaa agctctaact tcacagcccg tgagctggac 1080
ctcgaactgc agaaagatgt ggcacgcgtc attcttaccc aagtgggctc cggaccccaa 1140
gagacgaatg agtctctgat tgacgccaaa acgggtctcc ccaaagaa 1188
<210> 59
<211> 650
<212> PRT
<213> 解脂亚罗酵母（Yarrowia lipolytica）CLIB 122 / E 150
<400> 59
Met Ser Ala Asn Glu Asn Ile Ser Arg Phe Asp Ala Pro Val Gly Lys
1 5 10 15
Glu His Pro Ala Tyr Glu Leu Phe His Asn His Thr Arg Ser Phe Val
20 25 30
Tyr Gly Leu Gln Pro Arg Ala Cys Gln Gly Met Leu Asp Phe Asp Phe
35 40 45
Ile Cys Lys Arg Glu Asn Pro Ser Val Ala Gly Val Ile Tyr Pro Phe
50 55 60
Gly Gly Gln Phe Val Thr Lys Met Tyr Trp Gly Thr Lys Glu Thr Leu
65 70 75 80
Leu Pro Val Tyr Gln Gln Val Glu Lys Ala Ala Ala Lys His Pro Glu
85 90 95
Val Asp Val Val Val Asn Phe Ala Ser Ser Arg Ser Val Tyr Ser Ser
100 105 110
Thr Met Glu Leu Leu Glu Tyr Pro Gln Phe Arg Thr Ile Ala Ile Ile
115 120 125
Ala Glu Gly Val Pro Glu Arg Arg Ala Arg Glu Ile Leu His Lys Ala
130 135 140
Gln Lys Lys Gly Val Thr Ile Ile Gly Pro Ala Thr Val Gly Gly Ile
145 150 155 160
Lys Pro Gly Cys Phe Lys Val Gly Asn Thr Gly Gly Met Met Asp Asn
165 170 175
Ile Val Ala Ser Lys Leu Tyr Arg Pro Gly Ser Val Ala Tyr Val Ser
180 185 190
Lys Ser Gly Gly Met Ser Asn Glu Leu Asn Asn Ile Ile Ser His Thr
195 200 205
Thr Asp Gly Val Tyr Glu Gly Ile Ala Ile Gly Gly Asp Arg Tyr Pro
210 215 220
Gly Thr Thr Phe Ile Asp His Ile Leu Arg Tyr Glu Ala Asp Pro Lys
225 230 235 240
Cys Lys Ile Ile Val Leu Leu Gly Glu Val Gly Gly Val Glu Glu Tyr
245 250 255
Arg Val Ile Glu Ala Val Lys Asn Gly Gln Ile Lys Lys Pro Ile Val
260 265 270
Ala Trp Ala Ile Gly Thr Cys Ala Ser Met Phe Lys Thr Glu Val Gln
275 280 285
Phe Gly His Ala Gly Ser Met Ala Asn Ser Asp Leu Glu Thr Ala Lys
290 295 300
Ala Lys Asn Ala Ala Met Lys Ser Ala Gly Phe Tyr Val Pro Asp Thr
305 310 315 320
Phe Glu Asp Met Pro Glu Val Leu Ala Glu Leu Tyr Glu Lys Met Val
325 330 335
Ala Lys Gly Glu Leu Ser Arg Ile Ser Glu Pro Glu Val Pro Lys Ile
340 345 350
Pro Ile Asp Tyr Ser Trp Ala Gln Glu Leu Gly Leu Ile Arg Lys Pro
355 360 365
Ala Ala Phe Ile Ser Thr Ile Ser Asp Asp Arg Gly Gln Glu Leu Leu
370 375 380
Tyr Ala Gly Met Pro Ile Ser Glu Val Phe Lys Glu Asp Ile Gly Ile
385 390 395 400
Gly Gly Val Met Ser Leu Leu Trp Phe Arg Arg Arg Leu Pro Asp Tyr
405 410 415
Ala Ser Lys Phe Leu Glu Met Val Leu Met Leu Thr Ala Asp His Gly
420 425 430
Pro Ala Val Ser Gly Ala Met Asn Thr Ile Ile Thr Thr Arg Ala Gly
435 440 445
Lys Asp Leu Ile Ser Ser Leu Val Ala Gly Leu Leu Thr Ile Gly Thr
450 455 460
Arg Phe Gly Gly Ala Leu Asp Gly Ala Ala Thr Glu Phe Thr Thr Ala
465 470 475 480
Tyr Asp Lys Gly Leu Ser Pro Arg Gln Phe Val Asp Thr Met Arg Lys
485 490 495
Gln Asn Lys Leu Ile Pro Gly Ile Gly His Arg Val Lys Ser Arg Asn
500 505 510
Asn Pro Asp Phe Arg Val Glu Leu Val Lys Asp Phe Val Lys Lys Asn
515 520 525
Phe Pro Ser Thr Gln Leu Leu Asp Tyr Ala Leu Ala Val Glu Glu Val
530 535 540
Thr Thr Ser Lys Lys Asp Asn Leu Ile Leu Asn Val Asp Gly Ala Ile
545 550 555 560
Ala Val Ser Phe Val Asp Leu Met Arg Ser Cys Gly Ala Phe Thr Val
565 570 575
Glu Glu Thr Glu Asp Tyr Leu Lys Asn Gly Val Leu Asn Gly Leu Phe
580 585 590
Val Leu Gly Arg Ser Ile Gly Leu Ile Ala His His Leu Asp Gln Lys
595 600 605
Arg Leu Lys Thr Gly Leu Tyr Arg His Pro Trp Asp Asp Ile Thr Tyr
610 615 620
Leu Val Gly Gln Glu Ala Ile Gln Lys Lys Arg Val Glu Ile Ser Ala
625 630 635 640
Gly Asp Val Ser Lys Ala Lys Thr Arg Ser
645 650
<210> 60
<211> 1950
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 60
atgtctgcaa atgaaaacat ttcccgcttt gatgcacctg tgggtaagga gcatccagcc 60
tatgagctgt tccataacca cacccgctcc ttcgtctacg gtctccagcc tcgcgcgtgc 120
cagggtatgc tcgactttga tttcatttgt aagcgcgaga acccctcggt tgccggtgtt 180
atctaccctt tcggcggtca gttcgtgact aagatgtact ggggtaccaa ggagaccctg 240
ctcccggtgt accagcaagt tgaaaaagca gcagcgaaac atcccgaagt cgacgtcgtc 300
gtgaacttcg cgtcgtcccg cagcgtctac agctccacaa tggaactttt ggaataccca 360
caattccgca ccatcgctat catcgcagag ggcgtcccag aacgccgtgc gcgcgaaatc 420
ctgcacaagg ctcagaaaaa gggcgtgacg attatcggac cggcaaccgt cggcggcatt 480
aagcccggtt gctttaaagt tggtaatacg ggtggcatga tggacaacat cgtggcgtcc 540
aagctgtacc gcccaggttc cgttgcttac gtctccaagt ccggtggtat gtcaaatgag 600
cttaataaca tcatttccca cacgaccgat ggcgtgtacg agggaatcgc aattggtggc 660
gaccgctacc cgggcacaac ttttattgat cacatcttgc gttacgaagc ggatccaaag 720
tgcaagatca ttgtgctctt gggcgaagta ggtggcgtgg aagagtaccg cgttatcgag 780
gcggttaaaa acggtcaaat taagaagcca atcgtagcgt gggcaatcgg cacctgcgcc 840
tccatgttta agacggaagt acagtttggc catgctggct caatggctaa ctcagacctc 900
gagacggcta aagcaaaaaa tgcggcaatg aaatcagcag gcttttacgt gccggatacc 960
ttcgaggata tgcccgaagt cttggccgag ctctacgaga agatggtggc gaaaggcgag 1020
ctcagccgca tctccgaacc ggaggttcct aagatcccga tcgactatag ctgggcgcag 1080
gaacttggtt tgatccgcaa gccagccgca ttcatctcta ccatctcaga cgaccgcggc 1140
caagagctcc tttacgcggg catgcccatc tcagaggtct tcaaggagga catcggcatc 1200
ggtggcgtta tgtccctgct gtggtttcgt cgccgtctgc ctgattatgc atcgaaattc 1260
cttgaaatgg tgcttatgct gacggccgac cacggacctg ccgtttccgg tgcgatgaat 1320
acgatcatca ccactcgagc gggtaaggac ctcatttcct cccttgttgc cggccttctt 1380
acaattggta cccgcttcgg cggagcattg gacggcgcgg ccacagagtt tactaccgcc 1440
tacgataagg gcttgtcgcc ccgccaattt gtcgatacaa tgcgcaagca aaacaagctc 1500
atcccaggca tcggccaccg agtgaagtcc cgtaataacc ccgatttccg tgtggaactg 1560
gttaaggact ttgtgaagaa gaatttcccg tccacccagc tgttggatta cgcgctcgcc 1620
gttgaggagg tgaccacatc taaaaaggat aacttgatcc tcaacgttga tggtgcgatt 1680
gcggtttcgt tcgtcgatct tatgcgctcg tgtggtgctt ttaccgttga agaaacagaa 1740
gattatctga aaaatggcgt tttgaacggt ttgtttgtcc tcggtcgttc cattggcctg 1800
attgcgcacc acctggacca gaaacgtctt aaaaccggtc tttaccgcca tccttgggat 1860
gacattacgt acctcgttgg ccaagaggca attcagaaga agcgtgtgga gatctctgca 1920
ggcgacgttt caaaggcgaa gactcgaagc 1950
<210> 61
<211> 652
<212> PRT
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 61
Met Ser Gln Thr His Lys His Ala Ile Pro Ala Asn Ile Ala Asp Arg
1 5 10 15
Cys Leu Ile Asn Pro Glu Gln Tyr Glu Thr Lys Tyr Lys Gln Ser Ile
20 25 30
Asn Asp Pro Asp Thr Phe Trp Gly Glu Gln Gly Lys Ile Leu Asp Trp
35 40 45
Ile Thr Pro Tyr Gln Lys Val Lys Asn Thr Ser Phe Ala Pro Gly Asn
50 55 60
Val Ser Ile Lys Trp Tyr Glu Asp Gly Thr Leu Asn Leu Ala Ala Asn
65 70 75 80
Cys Leu Asp Arg His Leu Gln Glu Asn Gly Asp Arg Thr Ala Ile Ile
85 90 95
Trp Glu Gly Asp Asp Thr Ser Gln Ser Lys His Ile Ser Tyr Arg Glu
100 105 110
Leu His Arg Asp Val Cys Arg Phe Ala Asn Thr Leu Leu Asp Leu Gly
115 120 125
Ile Lys Lys Gly Asp Val Val Ala Ile Tyr Met Pro Met Val Pro Glu
130 135 140
Ala Ala Val Ala Met Leu Ala Cys Ala Arg Ile Gly Ala Val His Ser
145 150 155 160
Val Ile Phe Gly Gly Phe Ser Pro Glu Ala Val Ala Gly Arg Ile Ile
165 170 175
Asp Ser Ser Ser Arg Leu Val Ile Thr Ala Asp Glu Gly Val Arg Ala
180 185 190
Gly Arg Ser Ile Pro Leu Lys Lys Asn Val Asp Asp Ala Leu Lys Asn
195 200 205
Pro Asn Val Thr Ser Val Glu His Val Ile Val Leu Lys Arg Thr Gly
210 215 220
Ser Asp Ile Asp Trp Gln Glu Gly Arg Asp Leu Trp Trp Arg Asp Leu
225 230 235 240
Ile Glu Lys Ala Ser Pro Glu His Gln Pro Glu Ala Met Asn Ala Glu
245 250 255
Asp Pro Leu Phe Ile Leu Tyr Thr Ser Gly Ser Thr Gly Lys Pro Lys
260 265 270
Gly Val Leu His Thr Thr Gly Gly Tyr Leu Val Tyr Ala Ala Thr Thr
275 280 285
Phe Lys Tyr Val Phe Asp Tyr His Pro Gly Asp Ile Tyr Trp Cys Thr
290 295 300
Ala Asp Val Gly Trp Val Thr Gly His Ser Tyr Leu Leu Tyr Gly Pro
305 310 315 320
Leu Ala Cys Gly Ala Thr Thr Leu Met Phe Glu Gly Val Pro Asn Trp
325 330 335
Pro Thr Pro Ala Arg Met Cys Gln Val Val Asp Lys His Gln Val Asn
340 345 350
Ile Leu Tyr Thr Ala Pro Thr Ala Ile Arg Ala Leu Met Ala Glu Gly
355 360 365
Asp Lys Ala Ile Glu Gly Thr Asp Arg Ser Ser Leu Arg Ile Leu Gly
370 375 380
Ser Val Gly Glu Pro Ile Asn Pro Glu Ala Trp Glu Trp Tyr Trp Lys
385 390 395 400
Lys Ile Gly Lys Glu Lys Cys Pro Val Val Asp Thr Trp Trp Gln Thr
405 410 415
Glu Thr Gly Gly Phe Met Ile Thr Pro Leu Pro Gly Ala Ile Glu Leu
420 425 430
Lys Ala Gly Ser Ala Thr Arg Pro Phe Phe Gly Val Gln Pro Ala Leu
435 440 445
Val Asp Asn Glu Gly His Pro Gln Glu Gly Ala Thr Glu Gly Asn Leu
450 455 460
Val Ile Thr Asp Ser Trp Pro Gly Gln Ala Arg Thr Leu Phe Gly Asp
465 470 475 480
His Glu Arg Phe Glu Gln Thr Tyr Phe Ser Thr Phe Lys Asn Met Tyr
485 490 495
Phe Ser Gly Asp Gly Ala Arg Arg Asp Glu Asp Gly Tyr Tyr Trp Ile
500 505 510
Thr Gly Arg Val Asp Asp Val Leu Asn Val Ser Gly His Arg Leu Gly
515 520 525
Thr Ala Glu Ile Glu Ser Ala Leu Val Ala His Pro Lys Ile Ala Glu
530 535 540
Ala Ala Val Val Gly Ile Pro His Ala Ile Lys Gly Gln Ala Ile Tyr
545 550 555 560
Ala Tyr Val Thr Leu Asn His Gly Glu Glu Pro Ser Pro Glu Leu Tyr
565 570 575
Ala Glu Val Arg Asn Trp Val Arg Lys Glu Ile Gly Pro Leu Ala Thr
580 585 590
Pro Asp Val Leu His Trp Thr Asp Ser Leu Pro Lys Thr Arg Ser Gly
595 600 605
Lys Ile Met Arg Arg Ile Leu Arg Lys Ile Ala Ala Gly Asp Thr Ser
610 615 620
Asn Leu Gly Asp Thr Ser Thr Leu Ala Asp Pro Gly Val Val Glu Lys
625 630 635 640
Pro Leu Glu Glu Lys Gln Ala Ile Ala Met Pro Ser
645 650
<210> 62
<211> 1956
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 62
atgtctcaga ctcacaaaca cgctatccct gctaacattg ccgaccgctg tcttattaac 60
cctgagcaat atgaaactaa atacaagcaa tcgattaacg acccagatac cttttgggga 120
gagcagggca aaatcctcga ctggatcacc ccataccaaa aggtaaaaaa cacctcattt 180
gccccaggta acgtgtccat caagtggtac gaagatggta ctttgaacct ggcggccaac 240
tgccttgacc gacacctgca agagaacggt gaccgcaccg caatcatctg ggagggcgac 300
gacacctcac aatccaaaca catctcatat cgtgaacttc atcgcgacgt ttgccgtttc 360
gcaaacacgt tgctcgatct gggcatcaag aagggtgatg ttgtggcaat ctacatgccg 420
atggttccag aagcagcagt cgcgatgctt gcatgcgcac gcatcggcgc ggttcactca 480
gtgatcttcg gcggattttc ccccgaagcc gttgctggcc gtattatcga tagctcttcc 540
cgcctggtga tcactgctga cgagggtgtg cgcgcaggtc gttccattcc gttgaagaag 600
aacgtcgatg atgccttgaa gaatcccaat gtgacctccg tggaacacgt tatcgtgttg 660
aaacgcactg gctcagacat cgactggcag gaaggccgag atttgtggtg gcgagacctc 720
atcgaaaagg cgtccccgga acatcaacct gaggcaatga acgcggaaga tccgttgttt 780
attttgtaca cctcgggaag cacgggtaag ccaaaaggtg tgctccatac caccggcggt 840
tatctcgtgt acgcagcgac tactttcaaa tatgttttcg actatcaccc cggagatatc 900
tactggtgca cagcagacgt tggttgggtt accggacatt cctacctgct gtacggtcct 960
ctggcatgcg gcgcaactac gttgatgttt gaaggtgtgc ctaactggcc tactccagcg 1020
cgcatgtgtc aagtcgtgga caagcaccag gtaaacatcc tctatacagc acctaccgcc 1080
atccgagccc tcatggctga aggcgataag gccattgagg gtacagatcg ttcctccttg 1140
cgtatcctcg gctcagtggg cgaaccaatc aaccctgaag catgggaatg gtactggaag 1200
aagattggaa aggagaagtg tccagttgtg gatacctggt ggcagacaga gactggtggt 1260
ttcatgatta cccctctccc gggagcgatt gagttgaaag ccggttccgc gacccgccca 1320
ttcttcggag tacagccagc gcttgtggac aacgaaggac acccacagga aggcgccacc 1380
gaaggcaacc tggtcatcac ggacagctgg ccaggtcagg cacgtaccct gttcggcgac 1440
catgagcgct tcgagcaaac gtacttctct accttcaaga acatgtattt ctccggtgac 1500
ggtgcccgtc gcgatgaaga tggctactac tggattacag gccgcgttga tgacgttctc 1560
aacgttagcg gacaccgttt gggtaccgca gaaatcgaat ccgctctggt agcacacccg 1620
aaaatcgctg aagcagccgt ggtgggtatc ccgcacgcga tcaagggtca agctatctac 1680
gcatacgtca cgcttaatca cggagaagaa ccatcgcccg agttgtacgc cgaggtgcgc 1740
aattgggtcc gaaaagaaat tggcccactg gcaacccccg atgtgctcca ttggactgat 1800
agcctgccca agactcgttc tggtaaaatt atgcgccgca tcctgcgcaa gattgcagct 1860
ggagacacct ccaatttggg cgatacctcc actttggccg atccaggagt ggtggagaaa 1920
ccgctcgaag aaaaacaggc aatcgctatg ccttcc 1956
<210> 63
<211> 500
<212> PRT
<213> 酿酒酵母（S. cerevisiae）S288c
<400> 63
Met Thr Lys Leu His Phe Asp Thr Ala Glu Pro Val Lys Ile Thr Leu
1 5 10 15
Pro Asn Gly Leu Thr Tyr Glu Gln Pro Thr Gly Leu Phe Ile Asn Asn
20 25 30
Lys Phe Met Lys Ala Gln Asp Gly Lys Thr Tyr Pro Val Glu Asp Pro
35 40 45
Ser Thr Glu Asn Thr Val Cys Glu Val Ser Ser Ala Thr Thr Glu Asp
50 55 60
Val Glu Tyr Ala Ile Glu Cys Ala Asp Arg Ala Phe His Asp Thr Glu
65 70 75 80
Trp Ala Thr Gln Asp Pro Arg Glu Arg Gly Arg Leu Leu Ser Lys Leu
85 90 95
Ala Asp Glu Leu Glu Ser Gln Ile Asp Leu Val Ser Ser Ile Glu Ala
100 105 110
Leu Asp Asn Gly Lys Thr Leu Ala Leu Ala Arg Gly Asp Val Thr Ile
115 120 125
Ala Ile Asn Cys Leu Arg Asp Ala Ala Ala Tyr Ala Asp Lys Val Asn
130 135 140
Gly Arg Thr Ile Asn Thr Gly Asp Gly Tyr Met Asn Phe Thr Thr Leu
145 150 155 160
Glu Pro Ile Gly Val Cys Gly Gln Ile Ile Pro Trp Asn Phe Pro Ile
165 170 175
Met Met Leu Ala Trp Lys Ile Ala Pro Ala Leu Ala Met Gly Asn Val
180 185 190
Cys Ile Leu Lys Pro Ala Ala Val Thr Pro Leu Asn Ala Leu Tyr Phe
195 200 205
Ala Ser Leu Cys Lys Lys Val Gly Ile Pro Ala Gly Val Val Asn Ile
210 215 220
Val Pro Gly Pro Gly Arg Thr Val Gly Ala Ala Leu Thr Asn Asp Pro
225 230 235 240
Arg Ile Arg Lys Leu Ala Phe Thr Gly Ser Thr Glu Val Gly Lys Ser
245 250 255
Val Ala Val Asp Ser Ser Glu Ser Asn Leu Lys Lys Ile Thr Leu Glu
260 265 270
Leu Gly Gly Lys Ser Ala His Leu Val Phe Asp Asp Ala Asn Ile Lys
275 280 285
Lys Thr Leu Pro Asn Leu Val Asn Gly Ile Phe Lys Asn Ala Gly Gln
290 295 300
Ile Cys Ser Ser Gly Ser Arg Ile Tyr Val Gln Glu Gly Ile Tyr Asp
305 310 315 320
Glu Leu Leu Ala Ala Phe Lys Ala Tyr Leu Glu Thr Glu Ile Lys Val
325 330 335
Gly Asn Pro Phe Asp Lys Ala Asn Phe Gln Gly Ala Ile Thr Asn Arg
340 345 350
Gln Gln Phe Asp Thr Ile Met Asn Tyr Ile Asp Ile Gly Lys Lys Glu
355 360 365
Gly Ala Lys Ile Leu Thr Gly Gly Glu Lys Val Gly Asp Lys Gly Tyr
370 375 380
Phe Ile Arg Pro Thr Val Phe Tyr Asp Val Asn Glu Asp Met Arg Ile
385 390 395 400
Val Lys Glu Glu Ile Phe Gly Pro Val Val Thr Val Ala Lys Phe Lys
405 410 415
Thr Leu Glu Glu Gly Val Glu Met Ala Asn Ser Ser Glu Phe Gly Leu
420 425 430
Gly Ser Gly Ile Glu Thr Glu Ser Leu Ser Thr Gly Leu Lys Val Ala
435 440 445
Lys Met Leu Lys Ala Gly Thr Val Trp Ile Asn Thr Tyr Asn Asp Phe
450 455 460
Asp Ser Arg Val Pro Phe Gly Gly Val Lys Gln Ser Gly Tyr Gly Arg
465 470 475 480
Glu Met Gly Glu Glu Val Tyr His Ala Tyr Thr Glu Val Lys Ala Val
485 490 495
Arg Ile Lys Leu
500
<210> 64
<211> 1500
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 64
atgaccaagc ttcacttcga caccgcagaa ccggttaaga tcaccctgcc aaacggcctc 60
acttatgaac agcccaccgg cctgttcatc aacaacaaat tcatgaaggc gcaggatggc 120
aagacctacc cggtcgagga cccatcgacg gagaatactg tttgtgaagt gtcctcggcg 180
acgactgaag atgtcgagta tgcaatcgag tgcgctgatc gagcattcca tgatactgaa 240
tgggcgacac aagatcctcg tgagcgtggt cgtctgctct ccaagcttgc ggacgagctt 300
gaatcccaaa ttgatctggt gtcttctatt gaagctctcg ataatggaaa aaccctcgct 360
ctggcccgcg gtgacgtgac catcgctatt aactgcctgc gcgacgcggc agcttacgca 420
gataaggtca atggccgcac cattaacaca ggcgatggtt atatgaattt tacaaccttg 480
gagcctattg gagtgtgcgg ccagattatc ccatggaact tccccatcat gatgctcgct 540
tggaagattg ccccagccct tgcaatggga aacgtttgca ttctgaaacc ggccgcagtg 600
acgcccctga atgctctgta tttcgcatct ctgtgtaaga aggtaggtat ccccgctggc 660
gtcgttaaca ttgttcctgg tccaggtcgt actgtaggag cggcgctcac caacgatccg 720
cgcatccgta aactggcgtt caccggatcg acggaagtgg gtaagtccgt cgcagtcgac 780
tcttccgaat ctaatctcaa gaagattacg ctcgaactcg gcggcaaatc tgcacacctc 840
gtcttcgatg atgctaatat caagaaaacc ttgccgaacc tcgtcaacgg tattttcaag 900
aatgcgggcc aaatctgctc ctccggcagc cgtatctacg ttcaggaagg tatttacgac 960
gagctgctcg cagccttcaa ggcgtatctg gagacggaaa ttaaggtggg caacccattt 1020
gataaagcca actttcaggg cgctattact aatcgccagc agtttgacac cattatgaac 1080
tacattgata ttggtaagaa ggaaggcgct aagattctca ctggaggaga aaaggttggt 1140
gacaaaggat actttatccg tcccactgta ttctacgatg tgaacgaaga catgcgcatt 1200
gtgaaggaag aaattttcgg tccagttgtt acagtggcaa agttcaaaac cctcgaggaa 1260
ggtgttgaga tggctaatag cagcgagttc ggcctgggat caggcattga gacagaatcc 1320
ctttcgaccg gcctgaaggt ggccaagatg ttgaaggctg gcacagtttg gatcaatacc 1380
tataatgatt ttgactcccg tgttccgttt ggcggcgtga aacaatccgg ttacggccgc 1440
gaaatgggcg aggaggtgta ccacgcatac accgaagtga aggcagttcg aatcaaattg 1500
<210> 65
<211> 348
<212> PRT
<213> 酿酒酵母（S. cerevisiae）S288c
<400> 65
Met Ser Ile Pro Glu Thr Gln Lys Ala Ile Ile Phe Tyr Glu Ser Asn
1 5 10 15
Gly Lys Leu Glu His Lys Asp Ile Pro Val Pro Lys Pro Lys Pro Asn
20 25 30
Glu Leu Leu Ile Asn Val Lys Tyr Ser Gly Val Cys His Thr Asp Leu
35 40 45
His Ala Trp His Gly Asp Trp Pro Leu Pro Thr Lys Leu Pro Leu Val
50 55 60
Gly Gly His Glu Gly Ala Gly Val Val Val Gly Met Gly Glu Asn Val
65 70 75 80
Lys Gly Trp Lys Ile Gly Asp Tyr Ala Gly Ile Lys Trp Leu Asn Gly
85 90 95
Ser Cys Met Ala Cys Glu Tyr Cys Glu Leu Gly Asn Glu Ser Asn Cys
100 105 110
Pro His Ala Asp Leu Ser Gly Tyr Thr His Asp Gly Ser Phe Gln Glu
115 120 125
Tyr Ala Thr Ala Asp Ala Val Gln Ala Ala His Ile Pro Gln Gly Thr
130 135 140
Asp Leu Ala Glu Val Ala Pro Ile Leu Cys Ala Gly Ile Thr Val Tyr
145 150 155 160
Lys Ala Leu Lys Ser Ala Asn Leu Arg Ala Gly His Trp Ala Ala Ile
165 170 175
Ser Gly Ala Ala Gly Gly Leu Gly Ser Leu Ala Val Gln Tyr Ala Lys
180 185 190
Ala Met Gly Tyr Arg Val Leu Gly Ile Asp Gly Gly Pro Gly Lys Glu
195 200 205
Glu Leu Phe Thr Ser Leu Gly Gly Glu Val Phe Ile Asp Phe Thr Lys
210 215 220
Glu Lys Asp Ile Val Ser Ala Val Val Lys Ala Thr Asn Gly Gly Ala
225 230 235 240
His Gly Ile Ile Asn Val Ser Val Ser Glu Ala Ala Ile Glu Ala Ser
245 250 255
Thr Arg Tyr Cys Arg Ala Asn Gly Thr Val Val Leu Val Gly Leu Pro
260 265 270
Ala Gly Ala Lys Cys Ser Ser Asp Val Phe Asn His Val Val Lys Ser
275 280 285
Ile Ser Ile Val Gly Ser Tyr Val Gly Asn Arg Ala Asp Thr Arg Glu
290 295 300
Ala Leu Asp Phe Phe Ala Arg Gly Leu Val Lys Ser Pro Ile Lys Val
305 310 315 320
Val Gly Leu Ser Ser Leu Pro Glu Ile Tyr Glu Lys Met Glu Lys Gly
325 330 335
Gln Ile Ala Gly Arg Tyr Val Val Asp Thr Ser Lys
340 345
<210> 66
<211> 1044
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 66
atgtccatcc cggaaaccca gaaagccatt atcttttacg agagcaacgg caagctcgaa 60
cataaggata ttccagtgcc aaaaccgaag ccaaacgagc tcctgatcaa cgttaagtac 120
tccggagtgt gccacacgga tctgcacgct tggcatggcg attggcccct ccctactaag 180
ctgccactgg tcggcggaca cgaaggcgct ggtgtggtgg tgggaatggg tgagaacgtt 240
aaaggctgga agattggcga ctacgctggc atcaaatggc tgaatggttc ctgcatggct 300
tgtgaatact gtgagcttgg taatgaaagc aactgcccgc acgccgattt gagcggatac 360
acacatgatg gctccttcca ggagtacgct accgccgacg ctgtgcaagc agcgcacatt 420
ccacaaggca ccgacctcgc tgaagttgct ccaatcctgt gtgctggcat caccgtatac 480
aaggctctca aatccgcaaa cctgcgcgcg ggccactggg cagcgatttc aggcgctgcc 540
ggcggtctcg gttccttggc tgtgcaatac gcaaaggcta tgggataccg agtactggga 600
attgacggcg gaccaggaaa agaggaactt ttcacgtccc tcggcggcga ggtcttcatc 660
gattttacca aagagaagga catcgtctcc gctgtcgtga aggcaactaa cggcggcgct 720
cacggtatca tcaacgtgtc tgtttcggag gcggctatcg aggctagcac ccgatactgc 780
cgtgcaaatg gcacggtcgt gttggttggc ctgccagcgg gcgcaaagtg ctcttctgat 840
gtcttcaacc atgttgttaa gtccatttcg attgtgggct catacgttgg caatcgcgct 900
gacactcgtg aggcgttgga cttcttcgcg cgcggcctcg taaagtcgcc tatcaaggtt 960
gtcggtttga gcagcctgcc cgaaatctac gaaaaaatgg aaaagggtca aattgccggc 1020
cgatatgtcg tcgatacctc taaa 1044
<210> 67
<211> 497
<212> PRT
<213> 解脂亚罗酵母（Yarrowia lipolytica）CLIB 122 / E 150
<400> 67
Met Ser Ala Lys Ser Ile His Glu Ala Asp Gly Lys Ala Leu Leu Ala
1 5 10 15
His Phe Leu Ser Lys Ala Pro Val Trp Ala Glu Gln Gln Pro Ile Asn
20 25 30
Thr Phe Glu Met Gly Thr Pro Lys Leu Ala Ser Leu Thr Phe Glu Asp
35 40 45
Gly Val Ala Pro Glu Gln Ile Phe Ala Ala Ala Glu Lys Thr Tyr Pro
50 55 60
Trp Leu Leu Glu Ser Gly Ala Lys Phe Val Ala Lys Pro Asp Gln Leu
65 70 75 80
Ile Lys Arg Arg Gly Lys Ala Gly Leu Leu Val Leu Asn Lys Ser Trp
85 90 95
Glu Glu Cys Lys Pro Trp Ile Ala Glu Arg Ala Ala Lys Pro Ile Asn
100 105 110
Val Glu Gly Ile Asp Gly Val Leu Arg Thr Phe Leu Val Glu Pro Phe
115 120 125
Val Pro His Asp Gln Lys His Glu Tyr Tyr Ile Asn Ile His Ser Val
130 135 140
Arg Glu Gly Asp Trp Ile Leu Phe Tyr His Glu Gly Gly Val Asp Val
145 150 155 160
Gly Asp Val Asp Ala Lys Ala Ala Lys Ile Leu Ile Pro Val Asp Ile
165 170 175
Glu Asn Glu Tyr Pro Ser Asn Ala Thr Leu Thr Lys Glu Leu Leu Ala
180 185 190
His Val Pro Glu Asp Gln His Gln Thr Leu Leu Asp Phe Ile Asn Arg
195 200 205
Leu Tyr Ala Val Tyr Val Asp Leu Gln Phe Thr Tyr Leu Glu Ile Asn
210 215 220
Pro Leu Val Val Ile Pro Thr Ala Gln Gly Val Glu Val His Tyr Leu
225 230 235 240
Asp Leu Ala Gly Lys Leu Asp Gln Thr Ala Glu Phe Glu Cys Gly Pro
245 250 255
Lys Trp Ala Ala Ala Arg Ser Pro Ala Ala Leu Gly Gln Val Val Thr
260 265 270
Ile Asp Ala Gly Ser Thr Lys Val Ser Ile Asp Ala Gly Pro Ala Met
275 280 285
Val Phe Pro Ala Pro Phe Gly Arg Glu Leu Ser Lys Glu Glu Ala Tyr
290 295 300
Ile Ala Glu Leu Asp Ser Lys Thr Gly Ala Ser Leu Lys Leu Thr Val
305 310 315 320
Leu Asn Ala Lys Gly Arg Ile Trp Thr Leu Val Ala Gly Gly Gly Ala
325 330 335
Ser Val Val Tyr Ala Asp Ala Ile Ala Ser Ala Gly Phe Ala Asp Glu
340 345 350
Leu Ala Asn Tyr Gly Glu Tyr Ser Gly Ala Pro Asn Glu Thr Gln Thr
355 360 365
Tyr Glu Tyr Ala Lys Thr Val Leu Asp Leu Met Thr Arg Gly Asp Ala
370 375 380
His Pro Glu Gly Lys Val Leu Phe Ile Gly Gly Gly Ile Ala Asn Phe
385 390 395 400
Thr Gln Val Gly Ser Thr Phe Lys Gly Ile Ile Arg Ala Phe Arg Asp
405 410 415
Tyr Gln Ser Ser Leu His Asn His Lys Val Lys Ile Tyr Val Arg Arg
420 425 430
Gly Gly Pro Asn Trp Gln Glu Gly Leu Arg Leu Ile Lys Ser Ala Gly
435 440 445
Asp Glu Leu Asn Leu Pro Met Glu Ile Tyr Gly Pro Asp Met His Val
450 455 460
Ser Gly Ile Val Pro Leu Ala Leu Leu Gly Lys Arg Pro Lys Asn Val
465 470 475 480
Lys Pro Phe Gly Thr Gly Pro Ser Thr Glu Ala Ser Thr Pro Leu Gly
485 490 495
Val
<210> 68
<211> 1491
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 68
atgagcgcca agagcatcca cgaggctgat ggcaaagcac tccttgccca cttcttgtcc 60
aaagcaccag tctgggcaga gcaacagccc atcaatacct ttgagatggg tacccccaaa 120
ctggcaagcc tgactttcga ggatggtgta gcgcctgagc aaatcttcgc cgctgcagaa 180
aaaacctacc cgtggctgct tgaatcaggt gctaagttcg tcgccaagcc tgatcagctg 240
attaagcgac gtggtaaggc gggcttgctt gtgcttaaca aatcctggga agaatgtaag 300
ccctggattg ccgagcgcgc agcgaagcct attaatgtag agggaatcga cggcgtgttg 360
cgtacctttc tcgttgaacc atttgtcccc catgatcaga agcatgaata ttacatcaac 420
atccatagcg tccgcgaagg tgattggatc ctgttttacc acgaaggtgg tgtcgacgtg 480
ggcgacgttg atgctaaggc agcaaaaatc ctgatcccgg ttgatattga aaacgagtac 540
ccatcaaatg caaccctcac caaggagctt ttggctcatg tgcctgaaga tcagcaccaa 600
accttgctcg atttcatcaa ccgcctctac gcggtatacg tagatctgca attcacctac 660
ttggaaatta acccgctcgt ggtgattcct acggctcagg gagtcgaggt gcactatctc 720
gatttggcag gtaagttgga ccagaccgca gaattcgaat gtggacctaa atgggcggcc 780
gctcgttccc ccgcggcctt gggacaggtc gttacgatcg acgcgggtag caccaaggtc 840
tcgattgacg ctggtccagc catggtgttc ccagccccgt ttggccgcga actgagcaaa 900
gaagaagcat acatcgcgga actggattct aagacgggtg cctctctcaa gcttactgtc 960
ttgaacgcaa agggccgcat ctggaccctt gtcgcaggtg gaggagcaag cgtggtctac 1020
gcagacgcga tcgcctccgc tggatttgct gatgaactgg ctaattacgg agagtactcg 1080
ggtgcaccga acgagaccca gacttatgaa tatgccaaga ccgtcctcga tctcatgacc 1140
cgtggagacg cgcacccaga gggaaaggtg ctgttcatcg gcggaggcat cgccaacttc 1200
acccaggtag gctccacttt caaaggtatt atccgcgcct tccgtgacta ccaatcatcc 1260
cttcacaatc acaaggttaa gatctacgtc cgccgcggtg gtccaaactg gcaggaaggt 1320
ctgcgactta tcaaatctgc aggcgacgag ctgaatctcc caatggagat ttacggccca 1380
gatatgcacg tcagcggcat cgtgcccctc gctctgttgg gtaaacgccc taagaatgtg 1440
aagccattcg gaaccggccc ttccactgag gcatcgactc cactgggtgt g 1491
<210> 69
<211> 887
<212> PRT
<213> 大肠杆菌（E. coli）（菌株 K12）
<400> 69
Met Ser Glu Arg Phe Pro Asn Asp Val Asp Pro Ile Glu Thr Arg Asp
1 5 10 15
Trp Leu Gln Ala Ile Glu Ser Val Ile Arg Glu Glu Gly Val Glu Arg
20 25 30
Ala Gln Tyr Leu Ile Asp Gln Leu Leu Ala Glu Ala Arg Lys Gly Gly
35 40 45
Val Asn Val Ala Ala Gly Thr Gly Ile Ser Asn Tyr Ile Asn Thr Ile
50 55 60
Pro Val Glu Glu Gln Pro Glu Tyr Pro Gly Asn Leu Glu Leu Glu Arg
65 70 75 80
Arg Ile Arg Ser Ala Ile Arg Trp Asn Ala Ile Met Thr Val Leu Arg
85 90 95
Ala Ser Lys Lys Asp Leu Glu Leu Gly Gly His Met Ala Ser Phe Gln
100 105 110
Ser Ser Ala Thr Ile Tyr Asp Val Cys Phe Asn His Phe Phe Arg Ala
115 120 125
Arg Asn Glu Gln Asp Gly Gly Asp Leu Val Tyr Phe Gln Gly His Ile
130 135 140
Ser Pro Gly Val Tyr Ala Arg Ala Phe Leu Glu Gly Arg Leu Thr Gln
145 150 155 160
Glu Gln Leu Asp Asn Phe Arg Gln Glu Val His Gly Asn Gly Leu Ser
165 170 175
Ser Tyr Pro His Pro Lys Leu Met Pro Glu Phe Trp Gln Phe Pro Thr
180 185 190
Val Ser Met Gly Leu Gly Pro Ile Gly Ala Ile Tyr Gln Ala Lys Phe
195 200 205
Leu Lys Tyr Leu Glu His Arg Gly Leu Lys Asp Thr Ser Lys Gln Thr
210 215 220
Val Tyr Ala Phe Leu Gly Asp Gly Glu Met Asp Glu Pro Glu Ser Lys
225 230 235 240
Gly Ala Ile Thr Ile Ala Thr Arg Glu Lys Leu Asp Asn Leu Val Phe
245 250 255
Val Ile Asn Cys Asn Leu Gln Arg Leu Asp Gly Pro Val Thr Gly Asn
260 265 270
Gly Lys Ile Ile Asn Glu Leu Glu Gly Ile Phe Glu Gly Ala Gly Trp
275 280 285
Asn Val Ile Lys Val Met Trp Gly Ser Arg Trp Asp Glu Leu Leu Arg
290 295 300
Lys Asp Thr Ser Gly Lys Leu Ile Gln Leu Met Asn Glu Thr Val Asp
305 310 315 320
Gly Asp Tyr Gln Thr Phe Lys Ser Lys Asp Gly Ala Tyr Val Arg Glu
325 330 335
His Phe Phe Gly Lys Tyr Pro Glu Thr Ala Ala Leu Val Ala Asp Trp
340 345 350
Thr Asp Glu Gln Ile Trp Ala Leu Asn Arg Gly Gly His Asp Pro Lys
355 360 365
Lys Ile Tyr Ala Ala Phe Lys Lys Ala Gln Glu Thr Lys Gly Lys Ala
370 375 380
Thr Val Ile Leu Ala His Thr Ile Lys Gly Tyr Gly Met Gly Asp Ala
385 390 395 400
Ala Glu Gly Lys Asn Ile Ala His Gln Val Lys Lys Met Asn Met Asp
405 410 415
Gly Val Arg His Ile Arg Asp Arg Phe Asn Val Pro Val Ser Asp Ala
420 425 430
Asp Ile Glu Lys Leu Pro Tyr Ile Thr Phe Pro Glu Gly Ser Glu Glu
435 440 445
His Thr Tyr Leu His Ala Gln Arg Gln Lys Leu His Gly Tyr Leu Pro
450 455 460
Ser Arg Gln Pro Asn Phe Thr Glu Lys Leu Glu Leu Pro Ser Leu Gln
465 470 475 480
Asp Phe Gly Ala Leu Leu Glu Glu Gln Ser Lys Glu Ile Ser Thr Thr
485 490 495
Ile Ala Phe Val Arg Ala Leu Asn Val Met Leu Lys Asn Lys Ser Ile
500 505 510
Lys Asp Arg Leu Val Pro Ile Ile Ala Asp Glu Ala Arg Thr Phe Gly
515 520 525
Met Glu Gly Leu Phe Arg Gln Ile Gly Ile Tyr Ser Pro Asn Gly Gln
530 535 540
Gln Tyr Thr Pro Gln Asp Arg Glu Gln Val Ala Tyr Tyr Lys Glu Asp
545 550 555 560
Glu Lys Gly Gln Ile Leu Gln Glu Gly Ile Asn Glu Leu Gly Ala Gly
565 570 575
Cys Ser Trp Leu Ala Ala Ala Thr Ser Tyr Ser Thr Asn Asn Leu Pro
580 585 590
Met Ile Pro Phe Tyr Ile Tyr Tyr Ser Met Phe Gly Phe Gln Arg Ile
595 600 605
Gly Asp Leu Cys Trp Ala Ala Gly Asp Gln Gln Ala Arg Gly Phe Leu
610 615 620
Ile Gly Gly Thr Ser Gly Arg Thr Thr Leu Asn Gly Glu Gly Leu Gln
625 630 635 640
His Glu Asp Gly His Ser His Ile Gln Ser Leu Thr Ile Pro Asn Cys
645 650 655
Ile Ser Tyr Asp Pro Ala Tyr Ala Tyr Glu Val Ala Val Ile Met His
660 665 670
Asp Gly Leu Glu Arg Met Tyr Gly Glu Lys Gln Glu Asn Val Tyr Tyr
675 680 685
Tyr Ile Thr Thr Leu Asn Glu Asn Tyr His Met Pro Ala Met Pro Glu
690 695 700
Gly Ala Glu Glu Gly Ile Arg Lys Gly Ile Tyr Lys Leu Glu Thr Ile
705 710 715 720
Glu Gly Ser Lys Gly Lys Val Gln Leu Leu Gly Ser Gly Ser Ile Leu
725 730 735
Arg His Val Arg Glu Ala Ala Glu Ile Leu Ala Lys Asp Tyr Gly Val
740 745 750
Gly Ser Asp Val Tyr Ser Val Thr Ser Phe Thr Glu Leu Ala Arg Asp
755 760 765
Gly Gln Asp Cys Glu Arg Trp Asn Met Leu His Pro Leu Glu Thr Pro
770 775 780
Arg Val Pro Tyr Ile Ala Gln Val Met Asn Asp Ala Pro Ala Val Ala
785 790 795 800
Ser Thr Asp Tyr Met Lys Leu Phe Ala Glu Gln Val Arg Thr Tyr Val
805 810 815
Pro Ala Asp Asp Tyr Arg Val Leu Gly Thr Asp Gly Phe Gly Arg Ser
820 825 830
Asp Ser Arg Glu Asn Leu Arg His His Phe Glu Val Asp Ala Ser Tyr
835 840 845
Val Val Val Ala Ala Leu Gly Glu Leu Ala Lys Arg Gly Glu Ile Asp
850 855 860
Lys Lys Val Val Ala Asp Ala Ile Ala Lys Phe Asn Ile Asp Ala Asp
865 870 875 880
Lys Val Asn Pro Arg Leu Ala
885
<210> 70
<211> 2661
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 70
atgagcgagc gcttccccaa tgacgttgat cccattgaga cccgcgattg gttgcaggcg 60
atcgagtcgg tcatccgtga ggagggagtc gaacgagccc agtatttgat cgatcaactg 120
cttgcagaag cccgcaaggg aggcgtgaat gtcgcggctg gtaccggcat tagcaattac 180
atcaacacca tcccagtcga ggagcaacca gagtaccctg gtaatcttga actcgaacgc 240
cgtattcgca gcgccattcg ctggaacgca attatgaccg ttctgcgcgc gtcaaagaaa 300
gacctcgaac tgggaggtca tatggcatcc tttcaatcct cagccaccat ctacgacgtt 360
tgctttaacc acttcttccg cgcgcgtaat gaacaagatg gcggagacct ggtttatttt 420
cagggtcata tctcgccggg cgtctacgca cgcgccttcc tggaaggacg cctgacgcaa 480
gaacagctcg acaatttccg acaagaagtg cacggcaacg gactgtcctc ttacccgcat 540
ccaaaactga tgcctgagtt ctggcaattt ccaacggtgt ccatgggtct cggcccaatc 600
ggtgcaatct accaggcaaa gtttctgaag tacctggagc accgcggcct caaagatacg 660
tccaagcaaa ccgtctacgc gtttctgggt gacggagaaa tggatgagcc tgaatccaag 720
ggcgctatta cgattgccac tcgcgaaaag ctcgacaacc tggtcttcgt catcaattgc 780
aacctgcaac gattggacgg tccggttacg ggcaatggca agatcattaa tgagctcgaa 840
ggcatcttcg agggcgctgg ctggaacgtg attaaggtga tgtggggttc tcgctgggac 900
gaactgctgc gcaaagacac cagcggcaag ctgattcagc tgatgaacga gaccgtggat 960
ggtgattacc agaccttcaa gtccaaggat ggagcctacg tccgtgagca cttttttgga 1020
aaatatcctg aaacggcagc attggtggca gattggaccg acgagcagat ctgggcgctg 1080
aatcgcggtg gccatgaccc taagaaaatc tacgcggcct tcaagaaggc acaagaaacc 1140
aagggcaaag ccactgtcat cttggcgcat accatcaaag gctacggtat gggcgatgcc 1200
gctgagggca aaaacatcgc tcatcaagtt aagaagatga acatggatgg tgtccgccac 1260
atccgcgatc gctttaacgt ccccgtctct gacgcagata ttgagaagct gccatacatc 1320
accttcccag aaggctccga ggaacatacc tatctccacg cacagcgcca gaagctccat 1380
ggttaccttc cgtcgcgtca gccaaatttc accgaaaaat tggaattgcc ctcattgcaa 1440
gattttggcg cgctcttgga ggaacagtct aaagagatct ctacaaccat cgcgtttgtg 1500
cgtgccctca acgtcatgtt gaagaacaag tctatcaagg atcgattggt gcccattatc 1560
gctgatgaag cccgcacctt cggtatggag ggtttgtttc gccagattgg catctactct 1620
cccaatggcc agcaatacac cccacaggat cgcgaacaag ttgcctacta taaggaagac 1680
gagaaaggcc agatccttca agaaggcatc aacgaacttg gcgcgggatg cagctggctc 1740
gccgctgcca caagctactc aactaacaac ctcccgatga tccctttcta tatctactat 1800
tcgatgttcg gcttccaacg cattggcgac ttgtgctggg cggcaggcga tcaacaagcc 1860
cgcggctttc tgattggcgg tacttcgggt cgcactacgc tcaacggcga aggtctccag 1920
catgaggatg gtcattcgca catccaatca ctcaccattc caaactgtat ttcctatgac 1980
cccgcatacg cttacgaggt tgcggtcatc atgcacgatg gtctggaacg tatgtacggc 2040
gagaagcagg aaaatgtgta ttactacatc accaccctga atgaaaatta ccatatgcct 2100
gccatgcctg agggtgcgga agagggaatt cgtaagggta tctacaagct ggaaacaatt 2160
gaaggatcca agggtaaggt acagcttctg ggatctggct ccattcttcg tcatgtgcgc 2220
gaagcagcgg aaatcctcgc aaaagattac ggcgtaggca gcgacgttta ctccgttacg 2280
agcttcactg aactggcccg cgacggccag gactgtgagc gttggaacat gctgcatccc 2340
cttgaaaccc cacgcgtgcc gtatatcgca caagtaatga atgacgcccc tgccgtcgcc 2400
tcgactgact atatgaagct cttcgccgaa caggtccgca cgtacgttcc agcagatgat 2460
taccgcgttc tgggcaccga cggctttggc cgctccgatt cacgcgagaa cctgcgacac 2520
cacttcgagg tcgacgcttc ctacgtggta gtagctgcgc tcggagaact ggcaaagcgt 2580
ggcgagatcg acaaaaaggt agttgctgat gctatcgcta agttcaacat tgacgccgac 2640
aaggtcaatc cccgcttggc a 2661
<210> 71
<211> 120
<212> PRT
<213> 酿酒酵母（S. cerevisiae）S288c
<400> 71
Met Pro Lys Val Tyr Ser Tyr Gln Glu Val Ala Glu His Asn Gly Pro
1 5 10 15
Glu Asn Phe Trp Ile Ile Ile Asp Asp Lys Val Tyr Asp Val Ser Gln
20 25 30
Phe Lys Asp Glu His Pro Gly Gly Asp Glu Ile Ile Met Asp Leu Gly
35 40 45
Gly Gln Asp Ala Thr Glu Ser Phe Val Asp Ile Gly His Ser Asp Glu
50 55 60
Ala Leu Arg Leu Leu Lys Gly Leu Tyr Ile Gly Asp Val Asp Lys Thr
65 70 75 80
Ser Glu Arg Val Ser Val Glu Lys Val Ser Thr Ser Glu Asn Gln Ser
85 90 95
Lys Gly Ser Gly Thr Leu Val Val Ile Leu Ala Ile Leu Met Leu Gly
100 105 110
Val Ala Tyr Tyr Leu Leu Asn Glu
115 120
<210> 72
<211> 360
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 72
atgccaaagg tctattctta tcaggaagtc gcagaacaca acggcccaga aaacttctgg 60
atcattatcg atgataaggt ttacgatgtg tcgcagttca aagacgaaca cccaggtgga 120
gatgaaatta ttatggatct tggcggacaa gatgcgacgg aaagctttgt tgatatcggt 180
cactcagacg aagccttgcg cctcctcaag ggcctttaca tcggtgacgt agataaaacc 240
tcggagcgcg tttccgtcga gaaagtttcc acctcagaga accaatccaa aggctccggt 300
accctcgtgg tgatcctcgc tattctgatg ttgggtgtcg cctattatct tctgaatgag 360
<210> 73
<211> 485
<212> PRT
<213> 构巢裸胞壳（Emericella nidulans）ATCC 38163
<400> 73
Met Ser Ala Lys Ser Ile Phe Glu Ala Asp Gly Lys Ala Ile Leu Asn
1 5 10 15
Tyr His Leu Thr Arg Ala Pro Val Ile Lys Pro Thr Pro Leu Pro Pro
20 25 30
Ser Asn Thr His Asn Pro Pro Pro Lys Leu Ala Ser Leu Tyr Phe Pro
35 40 45
Asp Asp Leu Ser Val Lys Asp Val Leu Asp Gln Ala Glu Val Thr Tyr
50 55 60
Pro Trp Leu Leu Thr Pro Gly Ser Lys Phe Val Ala Lys Pro Asp Gln
65 70 75 80
Leu Ile Lys Arg Arg Gly Lys Ser Gly Leu Leu Ala Leu Asn Lys Thr
85 90 95
Trp Ala Glu Ala Arg Glu Trp Ile Glu Ala Arg Ala Thr Lys Glu Gln
100 105 110
Gln Val Glu Thr Val Val Gly Val Leu Arg His Phe Leu Val Glu Pro
115 120 125
Phe Val Pro His Pro Gln Glu Thr Glu Tyr Tyr Ile Asn Ile His Ser
130 135 140
Val Arg Glu Gly Asp Trp Ile Leu Phe Thr His Glu Gly Gly Val Asp
145 150 155 160
Val Gly Asp Val Asp Ala Lys Ala Glu Lys Leu Leu Ile Pro Val Asn
165 170 175
Leu Lys Asn Tyr Pro Ser Asn Glu Glu Ile Ala Ser Ala Leu Leu Ser
180 185 190
Lys Val Pro Lys Gly Ile His Asn Val Leu Val Asp Phe Ile Ser Arg
195 200 205
Leu Tyr Ala Val Tyr Val Asp Cys Gln Phe Thr Tyr Leu Glu Ile Asn
210 215 220
Pro Leu Val Val Ile Pro Asn Ala Asp Ala Thr Ser Ala Asp Val His
225 230 235 240
Phe Leu Asp Leu Ala Ala Lys Leu Asp Gln Thr Ala Glu Phe Glu Cys
245 250 255
Gly Thr Lys Trp Ala Val Ala Arg Ser Pro Ala Asn Leu Gly Leu Ala
260 265 270
Ala Leu Pro Thr Ser Asp Lys Val Asn Ile Asp Ala Gly Pro Pro Met
275 280 285
Glu Phe Pro Ala Pro Phe Gly Arg Glu Leu Ser Lys Glu Glu Lys Phe
290 295 300
Ile Ser Asp Met Asp Ala Lys Thr Gly Ala Ser Leu Lys Leu Thr Val
305 310 315 320
Leu Asn Pro Asn Gly Arg Val Trp Thr Leu Val Ala Gly Gly Gly Ala
325 330 335
Ser Val Val Tyr Ala Asp Ala Ile Ala Ser Ala Gly Phe Val Ser Glu
340 345 350
Leu Ala Asn Tyr Gly Glu Tyr Ser Gly Ala Pro Thr Glu Thr Gln Thr
355 360 365
Phe Asn Tyr Ala Arg Thr Ile Leu Asp Leu Met Leu Arg Ser Pro Ile
370 375 380
His Pro Asp Gly Lys Val Leu Phe Ile Gly Gly Gly Ile Ala Asn Phe
385 390 395 400
Thr Asn Val Ala Ser Thr Phe Lys Gly Val Ile Arg Ala Leu Arg Glu
405 410 415
Val Ala Pro Val Leu Asn Glu His Lys Val Gln Ile Trp Val Arg Arg
420 425 430
Ala Gly Pro Asn Tyr Gln Glu Gly Leu Lys Asn Ile Lys Ala Val Gly
435 440 445
Glu Glu Leu Gly Leu Asn Met His Val Tyr Gly Pro Glu Met His Val
450 455 460
Ser Gly Ile Val Pro Leu Ala Leu Gln Gly Lys Gln Thr Asp Ile Lys
465 470 475 480
Glu Phe Gly Thr Ala
485
<210> 74
<211> 1455
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 74
atgtcggcca agtcgatctt cgaagcagac ggaaaagcaa ttttgaatta ccacctgacc 60
cgcgcaccag tgatcaaacc aacccctctg ccaccttcca acactcataa cccccctccg 120
aaattggctt ctctctactt cccggacgac ttgtccgtga aggacgttct cgatcaagca 180
gaagttacat atccgtggct tcttacccca ggctccaaat ttgtggccaa acctgatcag 240
ttgatcaaac gtcgcggcaa atccggcctt ctggcgttga acaagacatg ggccgaagca 300
cgtgagtgga ttgaagctcg agcaactaag gaacaacagg tggaaactgt agtgggagtt 360
ctccgtcact tcctggttga accttttgtg cctcaccctc aggagacgga gtactacatt 420
aacatccact ccgtgcgcga gggtgactgg atcctcttta cccacgaggg cggtgtcgat 480
gtcggcgatg tcgacgccaa agcagagaag ctgttgatcc cggtgaatct caagaactac 540
ccctccaacg aggagatcgc atctgcattg ttgtccaagg tccctaaggg aatccataac 600
gtccttgtgg actttatctc ccgtctgtac gcggtctatg ttgattgtca atttacgtac 660
ctcgaaatca acccattggt cgtgattccc aacgcggacg caacttccgc tgatgttcat 720
tttcttgatc tcgctgcgaa gctggatcag acagccgaat tcgaatgcgg taccaagtgg 780
gccgtcgctc gttctcccgc caacttgggc ctcgcagcct tgcctacgag cgataaggtg 840
aacattgacg cgggacctcc gatggaattt cctgccccct ttggtcgcga gctgtcaaaa 900
gaagaaaaat ttatttccga catggatgct aagaccggag cgagccttaa gctcaccgtc 960
cttaatccga acggccgcgt gtggaccctc gtcgccggcg gaggtgcctc tgtggtatac 1020
gcagacgcga ttgcttctgc aggcttcgtg tccgaattgg ctaactacgg tgagtactcg 1080
ggcgcaccga ccgaaaccca aaccttcaac tacgcccgca ccatcttgga tctgatgctg 1140
cgctccccaa ttcacccgga cggtaaggtg cttttcattg gcggtggaat tgccaacttc 1200
acaaacgttg cgagcacctt caaaggagtg atccgtgctc tgcgtgaagt ggcaccagtt 1260
ttgaacgaac acaaagtgca gatctgggtg cgccgcgcag gtccaaatta ccaagaaggt 1320
ctgaaaaata tcaaagcggt tggtgaggaa ctcggactta acatgcatgt ctacggccca 1380
gaaatgcacg tcagcggcat tgtaccgctg gcactgcaag gcaagcaaac cgatattaaa 1440
gaattcggca ccgcg 1455
<210> 75
<211> 630
<212> PRT
<213> 大肠杆菌（E. coli）（菌株K12）
<400> 75
Met Ala Ile Glu Ile Lys Val Pro Asp Ile Gly Ala Asp Glu Val Glu
1 5 10 15
Ile Thr Glu Ile Leu Val Lys Val Gly Asp Lys Val Glu Ala Glu Gln
20 25 30
Ser Leu Ile Thr Val Glu Gly Asp Lys Ala Ser Met Glu Val Pro Ser
35 40 45
Pro Gln Ala Gly Ile Val Lys Glu Ile Lys Val Ser Val Gly Asp Lys
50 55 60
Thr Gln Thr Gly Ala Leu Ile Met Ile Phe Asp Ser Ala Asp Gly Ala
65 70 75 80
Ala Asp Ala Ala Pro Ala Gln Ala Glu Glu Lys Lys Glu Ala Ala Pro
85 90 95
Ala Ala Ala Pro Ala Ala Ala Ala Ala Lys Asp Val Asn Val Pro Asp
100 105 110
Ile Gly Ser Asp Glu Val Glu Val Thr Glu Ile Leu Val Lys Val Gly
115 120 125
Asp Lys Val Glu Ala Glu Gln Ser Leu Ile Thr Val Glu Gly Asp Lys
130 135 140
Ala Ser Met Glu Val Pro Ala Pro Phe Ala Gly Thr Val Lys Glu Ile
145 150 155 160
Lys Val Asn Val Gly Asp Lys Val Ser Thr Gly Ser Leu Ile Met Val
165 170 175
Phe Glu Val Ala Gly Glu Ala Gly Ala Ala Ala Pro Ala Ala Lys Gln
180 185 190
Glu Ala Ala Pro Ala Ala Ala Pro Ala Pro Ala Ala Gly Val Lys Glu
195 200 205
Val Asn Val Pro Asp Ile Gly Gly Asp Glu Val Glu Val Thr Glu Val
210 215 220
Met Val Lys Val Gly Asp Lys Val Ala Ala Glu Gln Ser Leu Ile Thr
225 230 235 240
Val Glu Gly Asp Lys Ala Ser Met Glu Val Pro Ala Pro Phe Ala Gly
245 250 255
Val Val Lys Glu Leu Lys Val Asn Val Gly Asp Lys Val Lys Thr Gly
260 265 270
Ser Leu Ile Met Ile Phe Glu Val Glu Gly Ala Ala Pro Ala Ala Ala
275 280 285
Pro Ala Lys Gln Glu Ala Ala Ala Pro Ala Pro Ala Ala Lys Ala Glu
290 295 300
Ala Pro Ala Ala Ala Pro Ala Ala Lys Ala Glu Gly Lys Ser Glu Phe
305 310 315 320
Ala Glu Asn Asp Ala Tyr Val His Ala Thr Pro Leu Ile Arg Arg Leu
325 330 335
Ala Arg Glu Phe Gly Val Asn Leu Ala Lys Val Lys Gly Thr Gly Arg
340 345 350
Lys Gly Arg Ile Leu Arg Glu Asp Val Gln Ala Tyr Val Lys Glu Ala
355 360 365
Ile Lys Arg Ala Glu Ala Ala Pro Ala Ala Thr Gly Gly Gly Ile Pro
370 375 380
Gly Met Leu Pro Trp Pro Lys Val Asp Phe Ser Lys Phe Gly Glu Ile
385 390 395 400
Glu Glu Val Glu Leu Gly Arg Ile Gln Lys Ile Ser Gly Ala Asn Leu
405 410 415
Ser Arg Asn Trp Val Met Ile Pro His Val Thr His Phe Asp Lys Thr
420 425 430
Asp Ile Thr Glu Leu Glu Ala Phe Arg Lys Gln Gln Asn Glu Glu Ala
435 440 445
Ala Lys Arg Lys Leu Asp Val Lys Ile Thr Pro Val Val Phe Ile Met
450 455 460
Lys Ala Val Ala Ala Ala Leu Glu Gln Met Pro Arg Phe Asn Ser Ser
465 470 475 480
Leu Ser Glu Asp Gly Gln Arg Leu Thr Leu Lys Lys Tyr Ile Asn Ile
485 490 495
Gly Val Ala Val Asp Thr Pro Asn Gly Leu Val Val Pro Val Phe Lys
500 505 510
Asp Val Asn Lys Lys Gly Ile Ile Glu Leu Ser Arg Glu Leu Met Thr
515 520 525
Ile Ser Lys Lys Ala Arg Asp Gly Lys Leu Thr Ala Gly Glu Met Gln
530 535 540
Gly Gly Cys Phe Thr Ile Ser Ser Ile Gly Gly Leu Gly Thr Thr His
545 550 555 560
Phe Ala Pro Ile Val Asn Ala Pro Glu Val Ala Ile Leu Gly Val Ser
565 570 575
Lys Ser Ala Met Glu Pro Val Trp Asn Gly Lys Glu Phe Val Pro Arg
580 585 590
Leu Met Leu Pro Ile Ser Leu Ser Phe Asp His Arg Val Ile Asp Gly
595 600 605
Ala Asp Gly Ala Arg Phe Ile Thr Ile Ile Asn Asn Thr Leu Ser Asp
610 615 620
Ile Arg Arg Leu Val Met
625 630
<210> 76
<211> 1890
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 76
atggccatcg aaatcaaggt gccagacatc ggtgccgatg aagtcgaaat caccgagatt 60
ctcgtcaaag tcggcgataa agtggaagca gaacagtcct tgatcaccgt agagggtgat 120
aaggctagca tggaagttcc tagcccgcag gctggcattg tgaaggaaat taaggtttca 180
gttggtgaca aaactcaaac gggagccctt attatgatct tcgactcggc agacggagct 240
gccgatgcgg cgccggccca agccgaagaa aagaaagaag cggctccagc agcagcaccc 300
gccgctgccg cagctaagga tgttaacgtg cctgatatcg gctctgacga agtggaagtc 360
accgaaatct tggtcaaggt gggtgataag gtggaagcag aacagtcttt gattacggtt 420
gaaggagata aggctagcat ggaagtgcca gcacccttcg cgggcaccgt taaggaaatc 480
aaagtgaatg tcggcgataa ggtatctacc ggctctctca ttatggtttt cgaagtcgcg 540
ggtgaagcag gagccgccgc accagccgca aagcaagagg cagcaccagc ggcagcacct 600
gctccggctg caggcgtgaa ggaggtaaac gtccctgata ttggtggcga tgaagttgag 660
gtgaccgagg tgatggtgaa agttggagat aaggttgcag ctgaacaatc cttgattact 720
gtggagggtg ataaggcgtc tatggaagtt cccgctccat ttgccggcgt cgtcaaggag 780
cttaaggtga acgttggaga caaggtgaag accggaagcc tcatcatgat tttcgaagtt 840
gagggtgcag cccctgccgc agctccagcc aagcaagagg cagccgctcc cgcgccagcg 900
gcgaaagctg aggctcctgc agcggccccc gccgctaaag ccgaaggcaa atccgaattc 960
gctgagaatg acgcttatgt acacgcaaca cctctgatcc gtcgcctggc acgcgaattt 1020
ggagtcaacc tcgcaaaggt caagggcaca ggccgcaagg gtcgaatcct tcgcgaagac 1080
gttcaagcat atgttaagga agccatcaag cgcgcggaag cggcaccggc agccaccggt 1140
ggtggaatcc ccggcatgtt gccctggccc aaagtggact tctcgaagtt tggcgagatt 1200
gaggaagtgg agctgggtcg catccagaaa atttccggcg caaacctgtc ccgcaactgg 1260
gtcatgattc ctcacgtgac ccatttcgat aaaaccgata ttaccgagtt ggaagcgttt 1320
cgtaaacaac aaaacgaaga agcggctaag cgtaagctcg acgttaaaat tacccccgtt 1380
gtctttatca tgaaggcggt agctgctgct ctcgaacaaa tgccgcgctt caactcttcc 1440
cttagcgagg acggacaacg ccttacattg aagaagtaca tcaatatcgg cgttgctgtc 1500
gatactccca acggcctcgt ggtcccagtg tttaaggatg ttaacaagaa gggtatcatt 1560
gagctcagcc gtgagctgat gacgatcagc aagaaggcac gcgatggtaa gctgaccgcg 1620
ggagaaatgc aaggtggctg cttcactatc tcgtctatcg gaggccttgg aacaacgcac 1680
ttcgcgccga tcgtcaacgc accagaggtg gcgatcctgg gcgtttccaa atctgcgatg 1740
gagcctgtct ggaatggcaa agagttcgta cctcgactta tgcttccaat ttccctgagc 1800
ttcgaccatc gcgtgatcga tggagcagat ggtgcacgct tcattaccat tatcaataac 1860
acgctgtcgg acatccgacg attggttatg 1890
<210> 77
<211> 352
<212> PRT
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 77
Met Ala Ser Glu Lys Glu Ile Arg Arg Glu Arg Phe Leu Asn Val Phe
1 5 10 15
Pro Lys Leu Val Glu Glu Leu Asn Ala Ser Leu Leu Ala Tyr Gly Met
20 25 30
Pro Lys Glu Ala Cys Asp Trp Tyr Ala His Ser Leu Asn Tyr Asn Thr
35 40 45
Pro Gly Gly Lys Leu Asn Arg Gly Leu Ser Val Val Asp Thr Tyr Ala
50 55 60
Ile Leu Ser Asn Lys Thr Val Glu Gln Leu Gly Gln Glu Glu Tyr Glu
65 70 75 80
Lys Val Ala Ile Leu Gly Trp Cys Ile Glu Leu Leu Gln Ala Tyr Trp
85 90 95
Leu Val Ala Asp Asp Met Met Asp Lys Ser Ile Thr Arg Arg Gly Gln
100 105 110
Pro Cys Trp Tyr Lys Val Pro Glu Val Gly Glu Ile Ala Ile Trp Asp
115 120 125
Ala Phe Met Leu Glu Ala Ala Ile Tyr Lys Leu Leu Lys Ser His Phe
130 135 140
Arg Asn Glu Lys Tyr Tyr Ile Asp Ile Thr Glu Leu Phe His Glu Val
145 150 155 160
Thr Phe Gln Thr Glu Leu Gly Gln Leu Met Asp Leu Ile Thr Ala Pro
165 170 175
Glu Asp Lys Val Asp Leu Ser Lys Phe Ser Leu Lys Lys His Ser Phe
180 185 190
Ile Val Thr Phe Lys Thr Ala Tyr Tyr Ser Phe Tyr Leu Pro Val Ala
195 200 205
Leu Ala Met Tyr Val Ala Gly Ile Thr Asp Glu Lys Asp Leu Lys Gln
210 215 220
Ala Arg Asp Val Leu Ile Pro Leu Gly Glu Tyr Phe Gln Ile Gln Asp
225 230 235 240
Asp Tyr Leu Asp Cys Phe Gly Thr Pro Glu Gln Ile Gly Lys Ile Gly
245 250 255
Thr Asp Ile Gln Asp Asn Lys Cys Ser Trp Val Ile Asn Lys Ala Leu
260 265 270
Glu Leu Ala Ser Ala Glu Gln Arg Lys Thr Leu Asp Glu Asn Tyr Gly
275 280 285
Lys Lys Asp Ser Val Ala Glu Ala Lys Cys Lys Lys Ile Phe Asn Asp
290 295 300
Leu Lys Ile Glu Gln Leu Tyr His Glu Tyr Glu Glu Ser Ile Ala Lys
305 310 315 320
Asp Leu Lys Ala Lys Ile Ser Gln Val Asp Glu Ser Arg Gly Phe Lys
325 330 335
Ala Asp Val Leu Thr Ala Phe Leu Asn Lys Val Tyr Lys Arg Ser Lys
340 345 350
<210> 78
<211> 1056
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 78
atggcatccg agaaggaaat ccgtcgtgaa cgattcctga acgtattccc aaagctcgtc 60
gaagagctta atgcttcctt gcttgcgtac ggaatgccga aggaagcgtg cgactggtat 120
gcgcattctc tcaactataa cacacctggc ggtaagctga atcgcggtct cagcgtggtg 180
gacacgtacg caatcctcag caataagacc gtggaacagc tgggacagga ggagtacgag 240
aaagttgcta tccttggatg gtgtatcgaa ctgctgcaag cctattggtt ggtagctgat 300
gacatgatgg ataagagcat cactcgacgt ggtcaaccat gctggtacaa ggtgccggag 360
gttggcgaaa tcgcgatctg ggatgcgttc atgttggagg ccgcaattta taagcttctc 420
aagtcccact ttcgtaacga aaagtactat attgacatca cagaactctt tcatgaagtg 480
accttccaaa ccgagctcgg ccaactcatg gatttgatca ccgcccccga agataaggtg 540
gacctttcga agttctccct caagaagcac tcgttcattg ttaccttcaa gacagcatac 600
tactcatttt acctgccagt cgccctcgca atgtacgtgg cgggcatcac tgatgaaaag 660
gatctgaagc aggcccgcga cgtccttatc cctcttggag agtacttcca gatccaggac 720
gattacttgg attgttttgg caccccagag cagattggta aaatcggtac tgatatccaa 780
gacaacaaat gttcctgggt aattaacaag gctctcgagt tggcttccgc ggaacaacgc 840
aagactcttg acgaaaatta cggtaagaaa gattcagtcg ctgaagccaa gtgcaagaaa 900
atctttaacg atcttaaaat cgaacagttg taccacgagt atgaagagtc tatcgctaaa 960
gacctgaaag ctaaaatttc ccaggtcgat gagtcacgcg gcttcaaagc ggatgtgctc 1020
accgccttcc ttaacaaggt gtataagcgc tccaaa 1056
<210> 79
<211> 299
<212> PRT
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 79
Met Asp Phe Pro Gln Gln Leu Glu Ala Cys Val Lys Gln Ala Asn Gln
1 5 10 15
Ala Leu Ser Arg Phe Ile Ala Pro Leu Pro Phe Gln Asn Thr Pro Val
20 25 30
Val Glu Thr Met Gln Tyr Gly Ala Leu Leu Gly Gly Lys Arg Leu Arg
35 40 45
Pro Phe Leu Val Tyr Ala Thr Gly His Met Phe Gly Val Ser Thr Asn
50 55 60
Thr Leu Asp Ala Pro Ala Ala Ala Val Glu Cys Ile His Ala Tyr Phe
65 70 75 80
Leu Ile His Asp Asp Leu Pro Ala Met Asp Asp Asp Asp Leu Arg Arg
85 90 95
Gly Leu Pro Thr Cys His Val Lys Phe Gly Glu Ala Asn Ala Ile Leu
100 105 110
Ala Gly Asp Ala Leu Gln Thr Leu Ala Phe Ser Ile Leu Ser Asp Ala
115 120 125
Asp Met Pro Glu Val Ser Asp Arg Asp Arg Ile Ser Met Ile Ser Glu
130 135 140
Leu Ala Ser Ala Ser Gly Ile Ala Gly Met Cys Gly Gly Gln Ala Leu
145 150 155 160
Asp Leu Asp Ala Glu Gly Lys His Val Pro Leu Asp Ala Leu Glu Arg
165 170 175
Ile His Arg His Lys Thr Gly Ala Leu Ile Arg Ala Ala Val Arg Leu
180 185 190
Gly Ala Leu Ser Ala Gly Asp Lys Gly Arg Arg Ala Leu Pro Val Leu
195 200 205
Asp Lys Tyr Ala Glu Ser Ile Gly Leu Ala Phe Gln Val Gln Asp Asp
210 215 220
Ile Leu Asp Val Val Gly Asp Thr Ala Thr Leu Gly Lys Arg Gln Gly
225 230 235 240
Ala Asp Gln Gln Leu Gly Lys Ser Thr Tyr Pro Ala Leu Leu Gly Leu
245 250 255
Glu Gln Ala Arg Lys Lys Ala Arg Asp Leu Ile Asp Asp Ala Arg Gln
260 265 270
Ser Leu Lys Gln Leu Ala Glu Gln Ser Leu Asp Thr Ser Ala Leu Glu
275 280 285
Ala Leu Ala Asp Tyr Ile Ile Gln Arg Asn Lys
290 295
<210> 80
<211> 897
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 80
atggacttcc cgcagcagct cgaggcctgc gttaagcagg ccaatcaggc gttgtcacgt 60
ttcatcgctc ccctgccttt ccagaacaca ccagtagtgg aaaccatgca atacggcgcc 120
ctgctgggtg gaaaacgctt gcgcccattc ttggtgtacg caaccggcca catgttcggc 180
gtctctacta acacccttga tgcacctgca gcggccgtgg agtgtatcca cgcctacttc 240
ttgatccacg acgatttgcc ggctatggat gatgatgatc tccgacgagg cctccctacc 300
tgccatgtaa aattcggcga agcgaatgcg atcttggcag gtgatgcgct gcagacactt 360
gccttttcga ttctgagcga cgcggatatg ccggaagtct ccgaccgaga ccgcatctcc 420
atgatcagcg agttggcttc tgcctccggt atcgcgggca tgtgcggagg tcaggctctt 480
gaccttgacg cagagggcaa gcacgttccc ttggacgctt tggagcgtat ccaccgccat 540
aagaccggcg ccttgatccg cgccgcggtc cgcttgggcg cgctgtctgc gggcgataag 600
ggacgccgcg cactcccagt gctcgataag tacgccgaat ccattggcct tgcttttcag 660
gtgcaggatg acatccttga tgtagtcggc gataccgcaa ctctcggcaa acgccagggc 720
gcggaccaac aattgggtaa gtcaacctac ccagctctgc tgggcttgga gcaagcacga 780
aagaaagccc gcgacctgat cgacgatgcc cgccaatctc ttaaacaact ggccgaacaa 840
tctctcgata ccagcgcact cgaggcactc gcagattaca tcatccaacg taataag 897
<210> 81
<211> 474
<212> PRT
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 81
Met Ser Thr Glu Ile Lys Thr Gln Val Val Val Leu Gly Ala Gly Pro
1 5 10 15
Ala Gly Tyr Ser Ala Ala Phe Arg Cys Ala Asp Leu Gly Leu Glu Thr
20 25 30
Val Ile Val Glu Arg Tyr Asn Thr Leu Gly Gly Val Cys Leu Asn Val
35 40 45
Gly Cys Ile Pro Ser Lys Ala Leu Leu His Val Ala Lys Val Ile Glu
50 55 60
Glu Ala Lys Ala Leu Ala Glu His Gly Ile Val Phe Gly Glu Pro Lys
65 70 75 80
Thr Asp Ile Asp Lys Ile Arg Thr Trp Lys Glu Lys Val Ile Asn Gln
85 90 95
Leu Thr Gly Gly Leu Ala Gly Met Ala Lys Gly Arg Lys Val Lys Val
100 105 110
Val Asn Gly Leu Gly Lys Phe Thr Gly Ala Asn Thr Leu Glu Val Glu
115 120 125
Gly Glu Asn Gly Lys Thr Val Ile Asn Phe Asp Asn Ala Ile Ile Ala
130 135 140
Ala Gly Ser Arg Pro Ile Gln Leu Pro Phe Ile Pro His Glu Asp Pro
145 150 155 160
Arg Ile Trp Asp Ser Thr Asp Ala Leu Glu Leu Lys Glu Val Pro Glu
165 170 175
Arg Leu Leu Val Met Gly Gly Gly Ile Ile Ala Leu Glu Met Ala Thr
180 185 190
Val Tyr His Ala Leu Gly Ser Gln Ile Asp Val Val Val Arg Lys His
195 200 205
Gln Val Ile Arg Ala Ala Asp Lys Asp Ile Val Lys Val Phe Thr Lys
210 215 220
Arg Ile Ser Lys Lys Phe Asn Leu Met Leu Glu Thr Lys Val Thr Ala
225 230 235 240
Val Glu Ala Lys Glu Asp Gly Ile Tyr Val Thr Met Glu Gly Lys Lys
245 250 255
Ala Pro Ala Glu Pro Gln Arg Tyr Asp Ala Val Leu Val Ala Ile Gly
260 265 270
Arg Val Pro Asn Gly Lys Asn Leu Asp Ala Gly Lys Ala Gly Val Glu
275 280 285
Val Asp Asp Arg Gly Phe Ile Arg Val Asp Lys Gln Leu Arg Thr Asn
290 295 300
Val Pro His Ile Phe Ala Ile Gly Asp Ile Val Gly Gln Pro Met Leu
305 310 315 320
Ala His Lys Gly Val His Glu Gly His Val Ala Ala Glu Val Ile Ala
325 330 335
Gly Lys Lys His Tyr Phe Asp Pro Lys Val Ile Pro Ser Ile Ala Tyr
340 345 350
Thr Glu Pro Glu Val Ala Trp Val Gly Leu Thr Glu Lys Glu Ala Lys
355 360 365
Glu Lys Gly Ile Ser Tyr Glu Thr Ala Thr Phe Pro Trp Ala Ala Ser
370 375 380
Gly Arg Ala Ile Ala Ser Asp Cys Ala Asp Gly Met Thr Lys Leu Ile
385 390 395 400
Phe Asp Lys Glu Ser His Arg Val Ile Gly Gly Ala Ile Val Gly Thr
405 410 415
Asn Gly Gly Glu Leu Leu Gly Glu Ile Gly Leu Ala Ile Glu Met Gly
420 425 430
Cys Asp Ala Glu Asp Ile Ala Leu Thr Ile His Ala His Pro Thr Leu
435 440 445
His Glu Ser Val Gly Leu Ala Ala Glu Val Phe Glu Gly Ser Ile Thr
450 455 460
Asp Leu Pro Asn Pro Lys Ala Lys Lys Lys
465 470
<210> 82
<211> 1422
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 82
atgtcgactg agatcaagac ccaagtcgta gtcctgggcg caggccctgc cggatactcc 60
gccgcgttcc gctgcgcgga cctcggtctt gaaaccgtga ttgtcgaacg ttacaacaca 120
ctgggcggtg tatgccttaa cgtgggctgc atcccctcca aagcactgtt gcacgttgcg 180
aaggtgattg aagaggcaaa ggcgctcgcg gaacacggca tcgtcttcgg cgaacctaag 240
actgacatcg ataaaattcg aacttggaag gaaaaagtga ttaaccagct taccggagga 300
ctggcaggta tggcaaaggg ccgaaaagtc aaagtggtga atggtttggg taagttcacc 360
ggcgccaaca ccctggaagt ggagggcgaa aacggcaaaa cggtcattaa tttcgacaac 420
gcaattattg cagcaggttc gcgccctatc cagctgccct tcattccaca tgaggacccc 480
cgtatctggg atagcacaga cgcactggaa ctcaaggaag tcccagaacg tctgctggtg 540
atgggtggcg gtattatcgc actcgaaatg gccacagtct accacgcgtt gggttctcag 600
atcgatgtgg ttgtacgcaa gcaccaagtg atccgcgccg cggacaaaga tatcgttaag 660
gtatttacca agcgcatttc aaagaagttc aacttgatgc tcgaaaccaa ggttaccgct 720
gttgaagcta aggaagacgg aatctacgtg accatggaag gaaagaaggc tcccgcagag 780
ccacagcgct acgatgcagt gcttgttgca attggacgcg tgcctaacgg caaaaatctc 840
gatgccggta aagctggcgt agaggtagat gaccgtggtt ttatccgcgt tgataagcaa 900
ttgcgaacga atgtgccaca catttttgct atcggtgaca ttgtgggtca accgatgctt 960
gcccacaagg gcgtccacga aggccacgtg gccgccgagg ttatcgccgg taagaaacac 1020
tactttgatc caaaagttat cccgtccatt gcctacactg aaccggaagt ggcatgggtg 1080
ggcttgaccg aaaaagaagc aaaagaaaag ggcattagct atgaaacagc aacctttccg 1140
tgggccgcat ccggccgcgc cattgcgagc gactgcgcgg acggcatgac taaattgatt 1200
tttgacaagg aaagccatcg cgtgatcgga ggtgcaatcg tgggtaccaa cggtggcgag 1260
ttgttgggtg agatcggcct cgctatcgaa atgggctgcg acgccgaaga tatcgctctg 1320
accattcacg cacacccaac cctgcatgaa tctgttggcc tggctgcgga ggtcttcgag 1380
ggctccatca ccgatctccc aaatccaaaa gcaaagaaaa aa 1422
<210> 83
<211> 474
<212> PRT
<213> 大肠杆菌（E. coli）ATCC 9637
<400> 83
Met Ser Thr Glu Ile Lys Thr Gln Val Val Val Leu Gly Ala Gly Pro
1 5 10 15
Ala Gly Tyr Ser Ala Ala Phe Arg Cys Ala Asp Leu Gly Leu Glu Thr
20 25 30
Val Ile Val Glu Arg Tyr Asn Thr Leu Gly Gly Val Cys Leu Asn Val
35 40 45
Gly Cys Ile Pro Ser Lys Ala Leu Leu His Val Ala Lys Val Ile Glu
50 55 60
Glu Ala Lys Ala Leu Ala Glu His Gly Ile Val Phe Gly Glu Pro Lys
65 70 75 80
Thr Asp Ile Asp Lys Ile Arg Thr Trp Lys Glu Lys Val Ile Asn Gln
85 90 95
Leu Thr Gly Gly Leu Ala Gly Met Ala Lys Gly Arg Lys Val Lys Val
100 105 110
Val Asn Gly Leu Gly Lys Phe Thr Gly Ala Asn Thr Leu Glu Val Glu
115 120 125
Gly Glu Asn Gly Lys Thr Val Ile Asn Phe Asp Asn Ala Ile Ile Ala
130 135 140
Ala Gly Ser Arg Pro Ile Gln Leu Pro Phe Ile Pro His Glu Asp Pro
145 150 155 160
Arg Ile Trp Asp Ser Thr Asp Ala Leu Glu Leu Lys Glu Val Pro Glu
165 170 175
Arg Leu Leu Val Met Gly Gly Gly Ile Ile Gly Leu Glu Met Gly Thr
180 185 190
Val Tyr His Ala Leu Gly Ser Gln Ile Asp Val Val Glu Met Phe Asp
195 200 205
Gln Val Ile Pro Ala Ala Asp Lys Asp Ile Val Lys Val Phe Thr Lys
210 215 220
Arg Ile Ser Lys Lys Phe Asn Leu Met Leu Glu Thr Lys Val Thr Ala
225 230 235 240
Val Glu Ala Lys Glu Asp Gly Ile Tyr Val Thr Met Glu Gly Lys Lys
245 250 255
Ala Pro Ala Glu Pro Gln Arg Tyr Asp Ala Val Leu Val Ala Ile Gly
260 265 270
Arg Val Pro Asn Gly Lys Asn Leu Asp Ala Gly Lys Ala Gly Val Glu
275 280 285
Val Asp Asp Arg Gly Phe Ile Arg Val Asp Lys Gln Leu Arg Thr Asn
290 295 300
Val Pro His Ile Phe Ala Ile Gly Asp Ile Val Gly Gln Pro Met Leu
305 310 315 320
Ala His Lys Gly Val His Glu Gly His Val Ala Ala Glu Val Ile Ala
325 330 335
Gly Lys Lys His Tyr Phe Asp Pro Lys Val Ile Pro Ser Ile Ala Tyr
340 345 350
Thr Glu Pro Glu Val Ala Trp Val Gly Leu Thr Glu Lys Glu Ala Lys
355 360 365
Glu Lys Gly Ile Ser Tyr Glu Thr Ala Thr Phe Pro Trp Ala Ala Ser
370 375 380
Gly Arg Ala Ile Ala Ser Asp Cys Ala Asp Gly Met Thr Lys Leu Ile
385 390 395 400
Phe Asp Lys Glu Ser His Arg Val Ile Gly Gly Ala Ile Val Gly Thr
405 410 415
Asn Gly Gly Glu Leu Leu Gly Glu Ile Gly Leu Ala Ile Glu Met Gly
420 425 430
Cys Asp Ala Glu Asp Ile Ala Leu Thr Ile His Ala His Pro Thr Leu
435 440 445
His Glu Ser Val Gly Leu Ala Ala Glu Val Phe Glu Gly Ser Ile Thr
450 455 460
Asp Leu Pro Asn Pro Lys Ala Lys Lys Lys
465 470
<210> 84
<211> 1422
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 84
atgtcaactg agattaaaac tcaggtggtc gttctgggcg ccggaccagc tggctattca 60
gcagcgtttc gctgcgcaga tctgggcctg gagacagtaa tcgttgagcg ttataacacc 120
cttggcggtg tctgcctgaa cgtcggatgc atcccttcta aggccctgct gcacgtggca 180
aaagtcatcg aagaagcgaa ggctcttgcc gaacacggca tcgtgtttgg agaaccaaag 240
actgacatcg acaagatccg cacatggaaa gagaaggtga tcaaccaact caccggtggt 300
ttggccggta tggcaaaggg tcgcaaagtt aaggtagtca atggccttgg caaatttacc 360
ggcgcaaaca ctcttgaagt ggaaggtgag aacggaaaga ctgtgatcaa cttcgacaac 420
gcaatcatcg cagccggctc gcgtcctatt caactgccgt tcatcccaca cgaagatccg 480
cgtatctggg atagcaccga tgcgctggag ctgaaagaag taccggaacg attgctcgtc 540
atgggtggag gtatcatcgg cttggagatg ggaaccgtat accacgcact tggctcccag 600
atcgacgttg tcgaaatgtt cgatcaggtg attccagcag cagataagga tatcgtgaag 660
gtgttcacca agcgcatttc aaagaagttc aatctgatgt tggaaaccaa ggttacggca 720
gtggaagcaa aggaggatgg tatctacgtg actatggagg gtaaaaaagc accagcggaa 780
ccccagcgat acgacgcagt tctcgtcgct atcggacgcg tcccaaatgg taagaatctg 840
gacgctggca aagcaggcgt cgaagtggat gaccgtggct tcatccgcgt agataaacag 900
ttgcgcacaa atgttccaca catctttgca attggagaca tcgtaggcca gcccatgctg 960
gcccacaaag gtgtccatga aggtcacgtg gcggctgagg ttatcgcagg caagaagcac 1020
tacttcgatc caaaggtgat ccccagcatt gcctatacgg agccagaggt agcttgggtt 1080
ggtttgaccg aaaaagaggc aaaggagaag ggtatctcgt atgaaacagc aacctttcca 1140
tgggccgcgt ccggacgtgc tatcgcaagc gactgcgcag atggcatgac caagctgatc 1200
ttcgacaaag aatcccaccg cgtgatcggt ggtgctattg ttggcactaa cggcggcgaa 1260
ctcttgggcg aaatcggctt ggcaattgaa atgggctgcg atgctgagga catcgctttg 1320
accatccacg cacacccgac cctgcatgag tctgtgggac tggcagccga agtgttcgaa 1380
ggctcaatca ccgacctccc taatcccaag gctaagaaga aa 1422
<210> 85
<211> 299
<212> PRT
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 85
Met Asp Phe Pro Gln Gln Leu Glu Ala Cys Val Lys Gln Ala Asn Gln
1 5 10 15
Ala Leu Ser Arg Phe Ile Ala Pro Leu Pro Phe Gln Asn Thr Pro Val
20 25 30
Val Glu Thr Met Gln Tyr Gly Ala Leu Leu Gly Gly Lys Arg Leu Arg
35 40 45
Pro Phe Leu Val Tyr Ala Thr Gly His Met Phe Gly Val Ser Thr Asn
50 55 60
Thr Leu Asp Ala Pro Ala Ala Ala Val Glu Cys Ile His Ala Tyr Phe
65 70 75 80
Leu Ile His Asp Asp Leu Pro Ala Met Asp Asp Asp Asp Leu Arg Arg
85 90 95
Gly Leu Pro Thr Cys His Val Lys Phe Gly Glu Ala Asn Ala Ile Leu
100 105 110
Ala Gly Asp Ala Leu Gln Thr Leu Ala Phe Ser Ile Leu Ser Asp Ala
115 120 125
Asp Met Pro Glu Val Ser Asp Arg Asp Arg Ile Ser Met Ile Ser Glu
130 135 140
Leu Ala Ser Ala Ser Gly Ile Ala Gly Met Cys Gly Gly Gln Ala Leu
145 150 155 160
Asp Leu Asp Ala Glu Gly Lys His Val Pro Leu Asp Ala Leu Glu Arg
165 170 175
Ile His Arg His Lys Thr Gly Ala Leu Ile Arg Ala Ala Val Arg Leu
180 185 190
Gly Ala Leu Ser Ala Gly Asp Lys Gly Arg Arg Ala Leu Pro Val Leu
195 200 205
Asp Lys Tyr Ala Glu Ser Ile Gly Leu Ala Phe Gln Val Gln Asp Asp
210 215 220
Ile Leu Asp Val Val Gly Asp Thr Ala Thr Leu Gly Lys Arg Gln Gly
225 230 235 240
Ala Asp Gln Gln Leu Gly Lys Ser Thr Tyr Pro Ala Leu Leu Gly Leu
245 250 255
Glu Gln Ala Arg Lys Lys Ala Arg Asp Leu Ile Asp Asp Ala Arg Gln
260 265 270
Ser Leu Lys Gln Leu Ala Glu Gln Ser Leu Asp Thr Ser Ala Leu Glu
275 280 285
Ala Leu Ala Asp Tyr Ile Ile Gln Arg Asn Lys
290 295
<210> 86
<211> 897
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 86
atggacttcc cgcaacagct tgaagcgtgt gtgaaacagg ccaaccaagc cctttcacgg 60
ttcattgccc cgcttccgtt ccaaaatacg ccggttgtag agacgatgca atacggcgcc 120
ctgcttggtg gtaaacggtt acgccctttc ctcgtgtacg caacaggaca tatgtttggt 180
gtcagcacaa atacactgga cgctccagcg gcggctgtgg aatgtatcca tgcatacttt 240
ttaattcacg atgacttgcc tgcaatggac gatgacgact taagacgtgg gctcccgacg 300
tgccacgtaa aatttggcga agctaacgcc attctggcgg gagacgcgct tcaaacgctc 360
gcctttagca tcctgtccga tgctgatatg ccggaggtat cggatcgcga tcgcattagt 420
atgatctcag aattggccag cgcttcaggg attgccggca tgtgtggcgg gcaggcgtta 480
gacttggacg cggagggcaa acacgtccct ttagatgcgc ttgagagaat acatagacat 540
aagacagggg cgcttatcag agcagctgta agacttggcg cgctgtctgc gggcgataaa 600
ggaagacggg ccctgccggt tttagataag tatgcagaat cgatcggact ggcgttccaa 660
gtgcaggacg acatccttga tgtcgttggt gataccgcga cacttgggaa aagacagggc 720
gcggatcaac agcttggcaa atccacctat ccggctttgc tcgggctgga gcaagcgcga 780
aaaaaagcta gagacttaat tgatgatgcc cgccagagct taaagcaact cgcggagcaa 840
tcgctggata catcagcact ggaagctctg gctgactata ttatccaacg taacaag 897
<210> 87
<211> 897
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 87
atggatttcc cacaacaatt ggaggcctgc gttaagcagg cgaaccaagc tctttcgcga 60
ttcatcgcgc cactcccgtt tcaaaacacg ccagtggtcg aaactatgca atacggcgct 120
ttgcttggtg gaaagcgttt gcgccccttc cttgtttacg caaccggtca catgtttggt 180
gtctctacta acacacttga tgcccccgcg gcggccgtgg agtgtatcca cgcttacttc 240
ctcattcacg atgacctccc tgcgatggac gatgatgacc tgcgtcgtgg cctgcccacc 300
tgtcacgtga aatttggtga agctaacgcc atcctggctg gcgacgcact gcagaccctt 360
gccttctcca ttctgagcga cgcagacatg cctgaagtct ccgaccgtga tcgcatctcg 420
atgatttccg agctcgcgtc cgccagcggc attgccggta tgtgcggtgg tcaggccctt 480
gacctggatg ctgaaggcaa acacgtgccg cttgatgctc tcgaacgcat ccaccgacac 540
aagaccggcg cgctgattcg tgctgccgtg cgcctcggcg ctttgtccgc aggtgacaaa 600
ggacgccgcg ctctcccagt attggataaa tatgccgaat ccattggtct cgccttccaa 660
gtgcaagacg atatcctcga cgtggtgggc gataccgcga cccttggaaa gcgtcaaggt 720
gcggaccagc agctgggaaa gtccacttat cccgcactcc tcggtttgga gcaggcacgc 780
aagaaggccc gcgacctgat tgatgacgcg cgtcaatcac tgaaacaact cgcagagcaa 840
tccctggata catctgccct ggaggccctg gcagactaca tcatccaacg taacaag 897
<210> 88
<211> 897
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 88
atggactttc cacaacagct ggaagcatgc gtgaagcagg ctaaccaggc tctatcaaga 60
ttcatcgcgc ccttaccttt ccaaaatacg cctgttgtgg aaactatgca gtatggggca 120
ttattgggtg gtaaaaggtt aaggccgttt ttagtgtatg ctacggggca catgttcggt 180
gtcagtacaa atacacttga cgcacctgca gcggctgttg aatgtattca tgcttatttt 240
ttaatccatg atgatttacc tgcaatggat gatgatgatt tgcgtagagg cctaccaaca 300
tgtcatgtga aatttggtga agcaaatgct attctagctg gggacgcttt gcaaacttta 360
gcgttcagca tcttgtccga tgcagatatg cctgaagtta gcgatagaga tagaattagt 420
atgatctcag aattagcatc cgcttctgga atcgctggca tgtgtggtgg ccaagcgctg 480
gatttggatg ctgaaggaaa acacgtaccg ttggatgcct tggaaagaat tcataggcat 540
aaaacgggtg cattgattcg tgctgctgtg cgtcttggtg ctctgtctgc gggagacaag 600
ggtagaaggg cgttgccagt tttggataaa tatgctgaaa gcattgggtt agcgtttcaa 660
gttcaagatg acatcttgga tgtggttgga gacacagcca ctttaggtaa aagacaagga 720
gcagaccagc aattaggcaa atctacttac cctgcattgt tgggtttaga gcaggctcgt 780
aagaaggcta gagacttgat agacgatgct agacagtcat tgaagcaatt ggccgaacag 840
agccttgata cgagcgccct tgaagcctta gctgattaca ttatccaaag aaataaa 897
<210> 89
<211> 897
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 89
atggactttc ctcagcaact ggaggcctgc gtgaagcagg ctaaccaggc actgagccgg 60
tttatcgccc cccttccctt tcaaaacact cctgtcgtcg agacgatgca atacggtgct 120
cttctcggtg gtaagcgact ccgacccttc ctcgtttacg ctacaggcca tatgtttgga 180
gtctccacca ataccctcga cgccccagct gccgccgtcg aatgcatcca tgcttacttc 240
ctgatccacg acgacctgcc cgctatggac gacgatgacc tgcgacgagg actccccact 300
tgtcatgtca aattcggcga ggctaacgcc attctggctg gagacgccct ccagactctc 360
gctttctcta tcctctccga cgctgacatg cctgaagtct ctgatcgaga ccggatttcc 420
atgatttccg agctggcctc ggcctccggc attgccggaa tgtgtggtgg tcaggctctc 480
gacctggacg ctgagggcaa gcatgtgccc ctcgacgccc ttgagcggat tcatcggcac 540
aagacgggcg ctctcatccg agctgccgtt cggctcggag ctctgtctgc cggagacaag 600
ggtcgacggg ctctccccgt ccttgataag tacgctgagt ccattggcct ggcctttcaa 660
gttcaggatg atattctcga cgtcgtcggt gatacggcca cccttggcaa gcgacaaggt 720
gctgaccagc agctgggcaa gtcgacctac cctgccctgc tcggactcga acaggcccga 780
aagaaggcac gtgatctgat cgacgacgca cgacagagcc tgaaacaact ggccgagcag 840
tctcttgaca cctccgccct ggaggctctc gccgactaca ttattcagcg aaacaag 897
<210> 90
<211> 1056
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 90
atggcatctg agaaggagat cagaagagaa cgctttttaa acgtgtttcc gaaactggta 60
gaagagttaa atgcgagcct gcttgcgtac ggcatgccaa aggaagcatg tgattggtat 120
gcacacagct taaattataa tacaccggga ggtaagctta acagagggct ttctgtagtg 180
gacacgtatg ctatattaag taataaaaca gtggaacaac ttggccaaga agaatatgaa 240
aaggtggcca tcctggggtg gtgcattgag cttttacaag catattggct tgtggcggat 300
gatatgatgg acaagtcaat tacacgtcgt ggtcagccgt gctggtataa ggtaccggaa 360
gttggggaaa tagctatctg ggacgcattt atgcttgaag ctgctattta taaacttctt 420
aaaagccatt tccgtaatga aaagtattac attgacatta ccgaactttt tcatgaggta 480
acatttcaaa ccgaactggg gcagcttatg gacttgatta cagcaccgga ggacaaagtt 540
gatctgtcga aattctcact taaaaaacat tcttttattg taacgttcaa gactgcgtat 600
tattcatttt atctgccggt cgcgcttgca atgtatgtag ccggcattac agatgaaaag 660
gacttaaaac aggctcggga tgttctgatt ccattggggg aatatttcca aattcaggat 720
gactatctgg actgttttgg taccccggaa cagattggta aaatcggcac agatatccaa 780
gataataaat gctcctgggt tatcaataaa gcactggaac tggctagcgc tgaacaaaga 840
aagactttgg atgaaaacta cggcaagaaa gatagcgtcg ctgaagctaa gtgcaaaaaa 900
atttttaacg acttaaaaat cgagcaactg taccacgaat atgaagaaag tattgctaaa 960
gatctcaaag ctaaaataag ccaggtggat gaatctcgcg ggttcaaagc agatgtattg 1020
acagcgtttc ttaataaagt gtataaacga tccaaa 1056
<210> 91
<211> 1056
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 91
atggctagcg aaaaggagat ccgaagagag agattcctca acgtgttccc taaacttgtg 60
gaggagctca acgcttcgct cctcgcttac ggtatgccca aggaggcctg tgattggtat 120
gcacactccc ttaactacaa cacgcctggt ggtaagctca accgaggtct ttctgttgtc 180
gatacctatg caattctttc caacaagacc gtcgagcaac tgggtcagga ggagtacgag 240
aaagtggcaa ttcttggctg gtgcattgaa ctcctccaag cttactggct ggtggctgat 300
gacatgatgg acaagagcat cacccgacga ggtcagccct gctggtacaa ggtgccagag 360
gttggagaga tcgccatttg ggacgcattt atgctggagg ctgccattta caagctcctt 420
aagtctcact ttcgaaacga gaagtactac atcgacatta ctgagctctt tcacgaagtg 480
acttttcaga ctgagctggg tcagcttatg gatctcatta ccgctcccga ggataaagtg 540
gatctgtcga agttttccct caagaagcat tcgtttattg tcaccttcaa gacagcatac 600
tactcttttt acctgcccgt cgccctcgca atgtacgtcg caggaattac cgatgagaaa 660
gacctcaagc aggcacgtga cgtcctcatc cccctcggcg agtactttca gattcaggat 720
gactatctcg actgcttcgg taccccagag caaatcggaa agattggaac tgacatccag 780
gacaataagt gctcttgggt tattaacaag gcccttgagc tggcttccgc cgagcaacga 840
aagacccttg acgagaatta cggcaagaag gatagcgtgg ccgaggccaa gtgcaagaag 900
atcttcaacg acctcaagat cgagcagctc taccacgagt acgaggagtc gattgctaag 960
gaccttaagg ctaagattag ccaggtggac gagtcccggg gattcaaggc tgacgtcctg 1020
accgcctttc tcaacaaggt ctacaagcga tccaag 1056
<210> 92
<211> 603
<212> PRT
<213> Perilla setoyensis
<400> 92
Met Cys Ser Ile Ser Gln Lys Val Val Ile Gly Leu Asn Lys Ala Ala
1 5 10 15
Ala Asn Asn Cys Leu Gln Asn Leu Asp Arg Arg Gly Phe Lys Thr Arg
20 25 30
Arg Val Ser Ser Ser Glu Ala Ala Ser Cys Leu Arg Ala Ser Ser Ser
35 40 45
Leu Gln Leu Asp Val Lys Pro Val Glu Glu Gly Arg Arg Ser Gly Asn
50 55 60
Tyr Gln Pro Ser Ile Trp Asp Phe Asn Tyr Val Gln Ser Leu Asn Thr
65 70 75 80
Pro Tyr Lys Glu Glu Arg Tyr Leu Thr Arg His Ala Glu Leu Ile Val
85 90 95
Gln Val Lys Pro Leu Leu Glu Lys Lys Met Glu Ala Thr Gln Gln Leu
100 105 110
Glu Leu Ile Asp Asp Leu Asn Asn Leu Gly Leu Ser Tyr Phe Phe Gln
115 120 125
Asp Arg Ile Lys Gln Ile Leu Ser Phe Ile Tyr Asp Glu Asn Gln Cys
130 135 140
Phe His Ser Asn Ile Asn Asp Gln Ala Glu Lys Arg Asp Leu Tyr Phe
145 150 155 160
Thr Ala Leu Gly Phe Arg Leu Leu Arg Gln His Gly Phe Asn Val Ser
165 170 175
Gln Glu Val Phe Asp Cys Phe Lys Asn Asp Lys Gly Ser Asp Phe Lys
180 185 190
Ala Ser Leu Ser Gly Asn Thr Lys Gly Leu Leu Gln Leu Tyr Glu Ala
195 200 205
Ser Phe Leu Val Arg Glu Gly Glu Asp Thr Leu Glu Leu Ala Arg Gln
210 215 220
Phe Ala Thr Lys Phe Leu Arg Arg Lys Leu Asp Glu Ile Asp Asp Asn
225 230 235 240
His Leu Leu Ser Arg Ile His His Ser Leu Glu Ile Pro Leu His Trp
245 250 255
Arg Ile Gln Arg Leu Glu Ala Arg Trp Phe Leu Asp Ala Tyr Ala Thr
260 265 270
Arg His Asp Met Asn Pro Ile Ile Leu Glu Leu Ala Lys Leu Asp Phe
275 280 285
Asn Ile Ile Gln Ala Thr His Gln Glu Glu Leu Lys Asp Val Ser Arg
290 295 300
Trp Trp Gln Asn Thr Arg Leu Ala Glu Lys Leu Pro Phe Val Arg Asp
305 310 315 320
Arg Leu Val Glu Ser Tyr Phe Trp Ala Ile Ala Leu Phe Glu Pro His
325 330 335
Gln Tyr Gly Tyr Gln Arg Arg Val Ala Ala Lys Ile Ile Thr Leu Ala
340 345 350
Thr Ser Ile Asp Asp Val Tyr Asp Ile Tyr Gly Thr Leu Asp Glu Leu
355 360 365
Gln Leu Phe Thr Asp Asn Phe Arg Arg Trp Asp Thr Glu Ser Leu Gly
370 375 380
Gly Leu Pro Tyr Ser Met Gln Leu Phe Tyr Met Val Ile His Asn Phe
385 390 395 400
Val Ser Glu Leu Ala Tyr Glu Ile Leu Lys Glu Lys Gly Phe Ile Ala
405 410 415
Ile Pro Tyr Leu Gln Arg Ser Trp Val Asp Leu Ala Glu Ser Phe Leu
420 425 430
Lys Glu Ala Asn Trp Tyr Tyr Ser Gly Tyr Thr Pro Ser Leu Glu Glu
435 440 445
Tyr Ile Asp Asn Gly Ser Ile Ser Ile Gly Ala Val Ala Val Leu Ser
450 455 460
Gln Val Tyr Phe Thr Leu Ala Asn Ser Ile Glu Lys Pro Lys Ile Glu
465 470 475 480
Ser Met Tyr Lys Tyr His His Ile Leu Arg Leu Ser Gly Leu Leu Val
485 490 495
Arg Leu His Asp Asp Leu Gly Thr Ser Leu Phe Glu Lys Lys Arg Gly
500 505 510
Asp Val Pro Lys Ala Val Glu Ile Cys Met Lys Glu Arg Asn Asp Thr
515 520 525
Glu Glu Glu Ala Glu Glu His Val Lys Tyr Leu Ile Arg Glu Ala Trp
530 535 540
Lys Glu Met Asn Thr Ala Thr Ala Ala Ala Gly Cys Pro Phe Met Asp
545 550 555 560
Glu Leu Asn Val Ala Ala Ala Asn Leu Gly Arg Ala Ala Gln Phe Val
565 570 575
Tyr Leu Asp Gly Asp Gly His Gly Val Gln His Ser Lys Ile His Gln
580 585 590
Gln Met Gly Gly Leu Met Phe Lys Pro Tyr Val
595 600
<210> 93
<211> 1809
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 93
atgtgcagca tctctcaaaa ggtcgttatt ggattgaata aggcagcggc caataactgc 60
ctccaaaacc ttgatcgccg aggttttaaa acacgccgtg tttcatcctc cgaagcggct 120
tcttgcctcc gagcctcaag ctcgctgcag cttgatgtca agccggtgga ggaaggccga 180
cgttccggta actatcagcc atcaatctgg gattttaatt atgtccagtc cctcaacacc 240
ccctacaaag aggaacgtta ccttacccga cacgcggaac tcatcgttca ggtcaagccg 300
cttctcgaaa aaaagatgga ggctactcag cagttggagc tcatcgatga ccttaacaac 360
ctcggtctga gctacttttt ccaagaccgc atcaagcaga ttctctcatt catctatgat 420
gaaaatcagt gtttccactc caatatcaat gatcaagctg aaaagcgaga cctgtacttt 480
accgcccttg gatttcgtct ccttcgccag cacggcttta acgtgtctca ggaggtgttc 540
gattgcttca aaaacgataa aggctccgat tttaaagcgt ccctgtctgg aaacaccaag 600
ggcctcctcc agctctacga agcctccttc ttggtccgtg agggtgaaga caccctcgaa 660
ctggcccgtc agttcgcgac caagttcctg cgccgcaagc ttgacgaaat cgacgacaac 720
catctgctct cacgaattca ccactcactc gaaatcccgt tgcactggcg catccagcgt 780
ctggaggcgc gctggtttct ggacgcgtac gctacacgcc acgatatgaa tcccattatc 840
ttggagctcg caaaactgga ctttaatatt attcaagcaa cccaccaaga agagctcaag 900
gacgtgagcc gttggtggca gaatacgcgc ctggccgaaa agctcccgtt tgtgcgcgat 960
cgacttgtcg agtcatactt ttgggccatc gcactctttg agccacatca atatggatac 1020
cagcgccgtg ttgctgccaa gattatcacc ctcgcgacat ccatcgatga cgtgtacgac 1080
atctacggca ctctcgacga gctccaactt tttaccgata acttccgtcg ctgggacacg 1140
gaatccttgg gcggtcttcc ttactcgatg cagttgttct acatggtgat tcacaacttt 1200
gtctctgagt tggcctatga aatcctgaaa gaaaaaggtt ttatcgccat cccctacctc 1260
caacgctcct gggttgattt ggcagaatcc tttttgaaag aggcaaactg gtactactct 1320
ggttatacac cctccctcga agaatacatc gacaacggct ccatctcaat tggcgcagta 1380
gctgtgttgt cccaagttta tttcactttg gcaaattcta tcgaaaagcc aaagattgaa 1440
tctatgtaca agtaccacca tattttgcgt ctgtcgggtt tgttggtgcg cctgcacgat 1500
gacttgggaa cttccttgtt cgagaagaag cgcggcgatg ttcctaaggc ggtcgaaatt 1560
tgcatgaaag aacgcaacga tacggaggag gaagcagaag aacacgttaa gtacttgatt 1620
cgcgaagcct ggaaggagat gaacactgct acggcagctg caggatgtcc ttttatggat 1680
gagctcaacg tggctgctgc caatctgggt cgcgcagccc agttcgtgta cctcgacgga 1740
gacggtcatg gcgtgcagca ctccaaaatc catcaacaga tgggtggcct tatgtttaaa 1800
ccgtatgtg 1809
<210> 94
<211> 1809
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 94
atgtgctcta tttcccaaaa agtagttatt ggccttaaca aggcggctgc taacaactgc 60
ttacaaaatc ttgacagacg tggtttcaaa actcgtagag tttcatcttc cgaggccgct 120
agctgcctgc gggcctctag ctctttgcag ctggacgtta aaccagtgga agaggggcgc 180
agatcgggga actatcagcc atcgatctgg gactttaatt acgtacaatc actgaatact 240
ccttacaaag aagaacggta tctgacacgg cacgcagaac tgattgtcca agtgaaacca 300
ctgctggaaa agaagatgga agccacccaa caactggaac tcatcgacga cctgaacaac 360
ttgggactgt cctacttttt tcaggacaga attaaacaaa ttctttcatt tatatacgac 420
gaaaatcaat gctttcattc taatatcaac gatcaagccg aaaaacggga cctttacttc 480
acggctttag gttttcgtct gcttagacag catggcttca atgtgagtca agaagtattc 540
gactgcttta aaaacgataa gggatcagac tttaaagctt cattatcagg taacaccaaa 600
ggcttgcttc aattgtatga agcgagtttt ttagtccgag aaggggaaga tactcttgaa 660
ttagcccgac agttcgcgac taaattcctg cgtagaaaac tggatgaaat cgacgataac 720
catcttttaa gccgcattca tcattccttg gaaattccgt tacattggcg cattcagcgg 780
ttggaagcac gctggttcct tgacgcttat gctacgcgtc atgacatgaa tccaatcatc 840
ctcgaactcg cgaaacttga ctttaatatt attcaagcta cacatcaaga agaacttaaa 900
gacgtatcta gatggtggca gaacacacgt ctggccgaga aacttccgtt tgtccgggac 960
agattagtgg aatcttattt ctgggctatt gctctttttg aacctcatca atatgggtac 1020
caacgtcgag ttgctgctaa gattattaca ttggcaacaa gcattgatga tgtttacgat 1080
atttacggga ctttggatga attacaactt tttacggata attttcggag atgggataca 1140
gagtccctgg gtgggcttcc gtattctatg cagctgtttt atatggttat ccataatttc 1200
gtgagcgaac ttgcgtatga aattctgaaa gaaaaaggct ttatagccat tccttatctt 1260
cagagaagtt gggttgacct ggccgagtcc tttctcaaag aggcgaactg gtactattcc 1320
ggctataccc ctagcctgga agaatacatc gataacgggt ctatcagcat aggcgcagtc 1380
gcagtgcttt cacaggttta ttttacgttg gctaacagta ttgaaaaacc taagatagag 1440
agcatgtata aatatcacca tatcttgcgc ttatctggtt tgttagtccg cttacatgat 1500
gaccttggaa caagtctgtt cgagaagaag cggggcgatg taccgaaggc ggttgagatt 1560
tgcatgaaag aacggaatga cacagaagaa gaagcagaag aacatgttaa atacctgatc 1620
cgcgaagctt ggaaagaaat gaacacagct acagcagctg ccgggtgtcc gtttatggat 1680
gaattgaatg tcgcggcggc aaatctgggt cgggcagccc agtttgtata tttggatggg 1740
gacggccatg gcgtccagca tagtaaaatt catcaacaga tgggaggctt gatgttcaaa 1800
ccttacgtc 1809
<210> 95
<211> 1809
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 95
atgtgttcaa tatcacaaaa agtcgtaatt gggttgaaca aggccgctgc caataactgt 60
ttgcagaatt tggataggag gggtttcaaa acaagaaggg tcagctcttc cgaggcggca 120
tcatgcttaa gagcttccag tagcttgcaa ctagatgtta aaccagtcga ggagggtaga 180
aggtcaggga actaccaacc aagtatctgg gatttcaatt atgtccaatc attgaatact 240
ccctataagg aggagcgtta cttaactaga catgcggaac tgattgtcca agtgaaacct 300
ttgttagaaa agaagatgga agctactcaa cagctggaat taattgacga tctaaataat 360
ttaggtttgt cttatttttt ccaagatagg atcaagcaaa tattgtcatt tatctatgac 420
gagaatcaat gtttccactc taacattaac gaccaggcag aaaagaggga cctgtatttt 480
accgcattgg gttttaggct tctacgtcag catggtttta atgtttctca agaagttttt 540
gattgcttta aaaatgacaa aggtagcgat tttaaggcta gcttatctgg caacaccaaa 600
ggtttgttgc aactatacga agcttctttt ctggtgagag aaggggaaga tactctagaa 660
ctagctagac aattcgctac caaatttctg aggagaaaat tggatgaaat tgatgataac 720
catctattaa gcagaataca ccattctcta gagattccat tgcattggag aatccaacgt 780
ttggaagcca ggtggttctt ggatgcttat gcaacacgtc atgatatgaa cccaattatt 840
ttggagcttg ccaaacttga ttttaacata attcaggcta cccatcaaga agaacttaag 900
gatgtttcca gatggtggca gaacactcgt ttggctgaaa agcttccatt tgtaagagat 960
agactagtcg aatcatactt ttgggcaatc gccttatttg aaccacatca gtacggttac 1020
caacgtagag tcgccgctaa aataataact ttggccactt ctatagatga tgtttatgat 1080
atttacggga cattagatga attgcaactt ttcaccgaca atttcagaag atgggatacc 1140
gaaagcctag gcggtctacc ttatagtatg caattgttct atatggttat tcataatttt 1200
gtttcagaat tggcttacga aatactgaaa gaaaaaggtt tcattgccat tccttattta 1260
caaaggagtt gggtggactt agcagaaagc tttctgaaag aggcgaattg gtattattca 1320
ggttatactc cgtctctgga ggagtatatt gataacggca gcatctctat tggtgcagtg 1380
gcagtattga gccaagtcta ctttacctta gctaacagca ttgaaaaacc aaaaattgag 1440
tccatgtata aataccatca catcttaaga ctttctggat tgttagtacg tttacacgac 1500
gatctgggca ccagtttgtt cgaaaaaaaa aggggcgacg tccctaaggc agtagaaatt 1560
tgtatgaaag aaaggaacga tacagaagaa gaagctgaag agcatgtgaa atatctgatt 1620
agagaggcgt ggaaggaaat gaacacagct acagctgctg cagggtgtcc attcatggat 1680
gaattaaacg tagctgcagc caatctgggt cgtgccgctc agttcgttta tttagacggt 1740
gatggtcatg gggttcagca ttccaagata caccaacaaa tggggggttt aatgtttaag 1800
ccttatgta 1809
<210> 96
<211> 1809
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 96
atgtgttcca tctcgcagaa agttgtgatc ggcctgaaca aggctgccgc taacaactgt 60
ctccagaacc tggatcgacg aggtttcaag acaagacggg tgtcttcttc cgaggctgct 120
tcctgcctgc gagcatctag ctcccttcag cttgacgtca agcctgtcga ggagggtcga 180
cggtctggca actaccaacc ctccatttgg gacttcaatt acgtgcaatc gctgaacact 240
ccttacaagg aggagcgata cctcacccga cacgccgagc tcattgttca ggtcaagccc 300
ctccttgaga agaagatgga ggcaacccag cagctggagc tgattgatga cctgaacaac 360
ctgggcctct cctacttctt tcaggaccgg attaagcaaa ttctgtcctt tatttacgac 420
gagaaccaat gttttcattc taacatcaac gaccaggctg aaaagcgaga tctttacttc 480
accgccctcg gattccgtct cctccgacag cacggcttta acgtttcgca agaggtcttt 540
gattgcttta agaatgataa aggctccgac ttcaaggctt ctctttctgg taacaccaag 600
ggtctgctgc aactgtacga agcttccttt cttgttagag agggcgagga taccctggag 660
ctggcccggc agtttgctac caagttcctg cgacgaaagc tggatgagat tgacgacaac 720
caccttctgt cccgaatcca ccactcgctt gagatccccc tccactggcg aatccagcga 780
ctggaagcta gatggtttct ggatgcctac gctactcgac acgacatgaa tcctattatc 840
ctggagctgg ccaagcttga cttcaacatc atccaggcca cccaccagga ggaactgaaa 900
gacgtctctc gatggtggca gaacacccga cttgccgaaa agctcccctt tgtgcgagac 960
cgactggtcg agagctactt ttgggctatt gccctctttg agccccacca gtacggttac 1020
cagcgaagag tcgctgctaa aatcatcacc ctggccacta gcatcgacga cgtttacgac 1080
atttacggaa ccctcgatga gctgcaactg tttaccgata actttcggcg ttgggacaca 1140
gaaagccttg gaggtctgcc ctactctatg cagctcttct atatggtgat ccacaacttt 1200
gtgtctgagc tggcttatga gatccttaag gaaaagggct tcatcgccat cccttacctt 1260
cagcgatcct gggtcgatct tgccgagtct ttcctcaagg aggccaactg gtattactct 1320
ggctacaccc catctctgga ggagtatatt gataacggtt ctatttctat cggcgccgtg 1380
gctgtccttt cccaagtcta tttcacactg gctaattcca tcgagaagcc caagattgaa 1440
tctatgtaca aataccatca catcctgcga ctgagcggtc ttctggtgcg gcttcatgac 1500
gatcttggca cttctctgtt cgagaagaaa cgaggagatg tgcctaaggc tgtcgagatt 1560
tgtatgaaag agcgtaacga caccgaggag gaggccgagg agcacgtgaa gtacctgatt 1620
cgagaggcat ggaaagaaat gaacaccgct actgccgccg ctggttgccc ctttatggat 1680
gagctgaacg ttgcagccgc caaccttggc cgagcagccc agttcgtcta cctggacggc 1740
gacggccacg gcgtccagca ttctaaaatt caccagcaga tgggtggcct gatgttcaag 1800
ccctatgtc 1809
<210> 97
<211> 591
<212> PRT
<213> 葡萄（Vitis vinifera）（葡萄）
<400> 97
Met Ser Arg Phe Val Thr Met Pro Ser His Val Leu Pro Ser Ser Phe
1 5 10 15
Val Ala Pro Ser Leu Gln Val Ser Ser Ser Pro Cys Ser Trp Arg Thr
20 25 30
Arg Pro Ser Pro Cys Thr Ser Cys His Leu Ser Pro Ser Ser Ser Ser
35 40 45
Lys Pro Leu Leu Gly Ser His Asp Tyr Ser Leu Leu Lys Ser Leu Thr
50 55 60
Leu Ser Pro His Ala Val Asn Ser Glu Ala Asp Ser Ser Thr Arg Arg
65 70 75 80
Met Lys Glu Val Lys Glu Arg Thr Trp Glu Ala Phe Tyr Arg Ala Trp
85 90 95
Asp Ser Arg Ala Ala Met Glu Met Val Asp Thr Val Glu Arg Leu Gly
100 105 110
Leu Ser Tyr His Phe Glu Asp Glu Ile Asn Ala Leu Leu Gln Arg Phe
115 120 125
Cys Asp Trp Asn Ala Ser Glu Asp Leu Phe Thr Thr Ala Leu Arg Phe
130 135 140
Arg Leu Leu Arg Gln Asn Gly Phe Pro Thr His Ser Asp Val Phe Gly
145 150 155 160
Lys Phe Met Asp Lys Asn Gly Lys Phe Lys Glu Ser Leu Thr Glu Asp
165 170 175
Ile Trp Gly Met Leu Ser Leu His Glu Ala Ser His Leu Gly Ala Lys
180 185 190
Asn Glu Glu Val Leu Ala Glu Ala Lys Glu Phe Thr Arg Ile His Leu
195 200 205
Ile Gln Ser Met Pro His Met Glu Pro His Phe Ser Ser His Val Gly
210 215 220
Arg Ala Leu Glu Leu Pro Arg His Leu Arg Met Val Arg Leu Glu Ala
225 230 235 240
Arg Asn Tyr Ile Gly Glu Tyr Ser Arg Glu Ser Asn Pro Asn Leu Ala
245 250 255
Phe Leu Glu Leu Ala Lys Leu Asp Phe Asp Met Val Gln Ser Leu His
260 265 270
Gln Lys Glu Leu Ala Glu Ile Val Arg Trp Trp Lys Gln Leu Gly Leu
275 280 285
Val Asp Lys Leu Asp Phe Ala Arg Asp Arg Pro Met Glu Cys Phe Leu
290 295 300
Trp Thr Val Gly Ile Phe Pro Asp Pro Arg His Ser Ser Cys Arg Ile
305 310 315 320
Glu Leu Thr Lys Ala Ile Ala Ile Leu Leu Val Ile Asp Asp Ile Tyr
325 330 335
Asp Ser Tyr Gly Ser Leu Asp Glu Leu Ala Leu Phe Thr Asp Ala Val
340 345 350
Lys Arg Trp Asp Leu Gly Ala Met Asp Gln Leu Pro Glu Tyr Met Lys
355 360 365
Ile Cys Tyr Met Ala Leu Tyr Asn Thr Thr Asn Asp Ile Ala Tyr Arg
370 375 380
Ile Leu Lys Glu His Gly Trp Ser Val Ile Glu Asp Leu Lys Arg Thr
385 390 395 400
Trp Met Asp Ile Phe Gly Ala Phe Leu Ala Glu Ala Tyr Cys Phe Lys
405 410 415
Gly Gly His Val Pro Ser Leu Glu Glu Tyr Leu Thr Asn Ala Val Thr
420 425 430
Thr Gly Gly Thr Tyr Met Ala Leu Val His Ala Phe Phe Leu Met Gly
435 440 445
Gln Gly Val Thr Arg Glu Asn Met Ala Met Leu Lys Pro Tyr Pro Asn
450 455 460
Ile Phe Ser Cys Ser Gly Lys Ile Leu Arg Leu Trp Asp Asp Leu Gly
465 470 475 480
Thr Ala Arg Glu Glu Gln Glu Arg Gly Asp Asn Ala Ser Ser Ile Glu
485 490 495
Cys Tyr Lys Arg Glu Arg Glu Met Asp Thr Val Leu Glu Asp Glu Ala
500 505 510
Cys Arg Lys His Ile Arg Gln Met Ile Gln Ser Leu Trp Val Glu Leu
515 520 525
Asn Gly Glu Leu Val Ala Ser Ser Ala Leu Pro Leu Ser Ile Ile Lys
530 535 540
Ala Ala Phe Asn Leu Ser Arg Thr Ala Gln Val Ile Tyr Gln His Gly
545 550 555 560
Asp Asp Asn Lys Thr Ser Ser Val Glu Asp His Val Gln Ala Leu Leu
565 570 575
Phe Arg Pro Val Ser Ser Asn Gly His Ala Gln Ile Thr Met His
580 585 590
<210> 98
<211> 1773
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 98
atgtctcgct tcgttacaat gccatctcat gtattaccta gttcattcgt tgcgcctagc 60
ctgcaggtgt cgtctagccc gtgttcttgg agaactcggc cgtccccttg cacttcatgc 120
catttatcac cgagctcttc atcaaaacct ctcttaggat ctcacgatta ttcacttttg 180
aaatccttaa cgttatctcc acacgccgta aactcggaag cagattcatc cacacggcgc 240
atgaaagagg tgaaagaaag aacgtgggaa gccttttatc gagcatggga tagtagagct 300
gccatggaga tggtggacac agtcgagcgc cttggacttt cctatcactt tgaagatgaa 360
attaatgcgc tgcttcaaag attttgcgac tggaacgcgt cagaggattt gtttacaacc 420
gccttaagat ttagactgct tcggcagaat gggtttccta ctcactccga tgtgtttgga 480
aagtttatgg acaagaacgg aaaatttaag gagagtctga cggaagatat ttggggtatg 540
ctttcgcttc atgaagcttc acatcttggg gcaaaaaacg aggaagtcct tgctgaagca 600
aaagagttta ctcgaataca tcttattcag tctatgccgc atatggagcc tcatttctcg 660
tcacacgtag gtcgggcact tgaactccct cgtcatctcc ggatggtgcg tctcgaagct 720
cgtaattata tcggggaata ttcacgtgaa agcaacccga acctcgcatt ccttgaattg 780
gccaaattgg atttcgatat ggtacaatca ttgcatcaaa aagagctggc ggaaattgtt 840
cgatggtgga aacaattagg attagtcgac aaattagatt ttgctcgcga tcggccgatg 900
gagtgttttc tctggaccgt tggtatcttc ccggatcctc ggcattccag ctgtcgtatt 960
gaattgacga aagctattgc gattctgttg gtcattgacg atatttatga ttcatatgga 1020
agcttggatg aactggcttt atttactgat gctgttaaaa gatgggatct gggtgcgatg 1080
gatcaacttc cggagtatat gaaaatctgc tacatggctc tgtacaacac aacaaacgat 1140
atagcataca gaatattaaa agagcacggc tggagcgtaa ttgaagattt aaaacgcaca 1200
tggatggaca tctttggggc gtttttggca gaagcatact gtttcaaagg tggacatgtt 1260
ccgagccttg aggaatactt aacaaacgca gtcacgactg gcggcaccta tatggccctt 1320
gttcacgcgt tctttcttat gggacaaggc gtgactcgtg aaaatatggc catgcttaaa 1380
ccgtatccta atatcttctc atgtagcggt aaaattcttc gactttggga tgacttgggt 1440
acagctcgag aagaacaaga gcgcggggat aatgcgagct ccattgaatg ctacaaaaga 1500
gaaagagaaa tggatacggt actggaagat gaagcatgta gaaaacatat cagacagatg 1560
atccaatcac tgtgggtgga acttaacggc gagctggtgg ccagcagcgc tctgccgttg 1620
tcaattataa aagccgcgtt taacctttcc cgtacagctc aagttatcta tcagcacggt 1680
gatgataaca aaacatcatc agttgaagac catgtccaag cccttttgtt tcgtccggta 1740
tcttcaaatg gtcatgctca aatcacgatg cat 1773
<210> 99
<211> 1773
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 99
atgtcccgtt ttgtcactat gccatcgcac gtcttgccta gcagcttcgt agcgccatct 60
ctgcaagtgt cttcatcccc atgctcgtgg cgcacccgcc cttccccatg cacctcttgt 120
cacttgtcac catcctcttc ttcaaaacca cttttgggca gccacgatta ctccctgctt 180
aagagcctga ccctttctcc acacgccgtg aattccgaag cggattcttc cacccgccga 240
atgaaggaag tcaaggagcg cacatgggaa gctttctatc gagcctggga ttcacgtgct 300
gctatggaga tggttgacac cgtggaacgt ctgggtcttt cataccactt tgaggacgaa 360
attaatgcgc tcttgcagcg cttttgtgat tggaacgctt ccgaagatct gttcacaacc 420
gccttgcgtt tccgccttct gcgccagaac ggttttccta cccattctga cgtctttggc 480
aaattcatgg ataagaacgg aaaattcaag gaaagcttga ctgaggatat ctggggaatg 540
ctgtcattgc atgaggcatc gcacttgggc gcaaaaaacg aggaggtcct cgcggaagca 600
aaggagttca cccgcatcca tctgatccaa tcaatgccgc atatggaacc tcacttctcc 660
tcccacgtcg gacgcgctct ggaactccct cgccatcttc gcatggtgcg actcgaagct 720
cgcaattata tcggcgagta ctcccgcgag agcaacccga atcttgcatt cttggaactc 780
gcgaagttgg attttgacat ggtgcagtca ctccaccaaa aggaactcgc ggagatcgtt 840
cgctggtgga aacagctggg tctcgtggac aagttggatt tcgcgcgcga ccgacctatg 900
gaatgcttcc tctggaccgt gggcatcttt cctgatcctc gacattcctc ttgtcgcatc 960
gaattgacta aagcaatcgc catcctcctg gttatcgacg atatttatga ctcatatggc 1020
agcctcgacg aattggcact gttcacagac gcagtcaaac gctgggattt gggagcgatg 1080
gatcagttgc ccgaatatat gaagatctgc tatatggcac tttacaacac cacaaacgat 1140
atcgcgtacc gcatccttaa agaacatggt tggtccgtta tcgaagactt gaagcgcacc 1200
tggatggaca tcttcggtgc gttcctcgcg gaggcatact gtttcaaagg cggtcacgta 1260
ccctctctcg aggaatacct caccaatgca gtcaccactg gaggcactta tatggccctg 1320
gtccacgctt ttttcctcat gggtcagggc gtgacccgcg aaaatatggc catgcttaag 1380
ccatacccaa acattttctc atgttccgga aaaattctgc gcctttggga cgatcttggt 1440
accgctcgcg aggagcaaga acgcggagac aacgcttctt ccatcgagtg ctataaacga 1500
gagcgcgaaa tggacaccgt gctcgaagac gaagcatgtc gcaagcatat ccgccaaatg 1560
atccagtcac tttgggtgga gcttaacggt gagctcgtgg catcttcggc tcttccactg 1620
tccattatta aggcggcgtt caacctgtct cgcacggcac aagtcatcta ccaacacgga 1680
gacgataaca agacttcgtc tgtggaggac catgtacagg ccttgctgtt tcgtcccgtt 1740
tcttcaaacg gccacgctca aatcaccatg cac 1773
<210> 100
<211> 1773
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 100
atgagtagat ttgttactat gccctcacat gtcctaccat cttcttttgt agccccttcc 60
ttgcaagtca gctcaagtcc gtgtagttgg agaactaggc cgagcccatg tacttcatgt 120
catttaagtc cgtcctcaag cagcaaacca ttgctgggat cccatgacta ttctttgcta 180
aaatccttga cgttgtctcc tcacgcagtt aactcagagg cagattcatc tacaaggaga 240
atgaaagaag tgaaagaaag aacttgggaa gcgttctaca gagcatggga ttcaagggca 300
gcaatggaaa tggtcgatac ggtggaaaga ttaggcttgt cataccattt tgaagacgaa 360
ataaatgctt tgcttcaaag gttctgtgac tggaatgcat ctgaggatct ttttacgacc 420
gcactaagat ttagactgtt gagacagaac ggcttcccaa cgcatagcga tgtgtttggt 480
aagttcatgg ataagaatgg gaaatttaag gagtccttaa ccgaagacat ctggggaatg 540
ttatccttgc atgaagcttc ccaccttggt gccaaaaacg aggaagtctt ggccgaagct 600
aaagaattca ccagaattca cctaatacaa tccatgcccc atatggaacc acatttctcc 660
tctcatgtgg ggagagcctt agagttgccg agacacttaa ggatggttag actagaagct 720
aggaactata tcggggaata ttcaagagaa agtaatccga atttggcgtt tttggagctt 780
gctaagttgg attttgatat ggttcaatcc ctgcaccaaa aagaactagc cgaaatagtt 840
aggtggtgga agcaattagg tctggttgat aaactggact ttgctagaga tagaccaatg 900
gagtgctttc tttggacggt aggtatcttc cctgacccaa ggcacagctc ttgtagaatt 960
gaacttacta aggcaatcgc catcctattg gtaattgatg atatctacga tagttatggt 1020
agtctggacg aattggcctt atttactgac gcagtaaaga gatgggatct aggtgctatg 1080
gatcagctgc ctgagtatat gaaaatctgt tacatggccc tttataacac cactaatgat 1140
atcgcatata gaatcttgaa ggagcacggt tggtcagtga ttgaagattt aaaaaggact 1200
tggatggata tttttggagc ttttcttgct gaagcttatt gctttaaagg tggtcatgtc 1260
ccctcattag aagaatacct gaccaatgcc gtgactactg gtggtaccta catggcttta 1320
gtgcatgcct tttttcttat gggtcaaggc gttactcgtg aaaacatggc aatgctgaaa 1380
ccatatccca atatattcag ctgctcaggt aagattttac gtttatggga cgatcttggt 1440
actgcaagag aagaacaaga aaggggggac aatgcatcca gcattgaatg ctataaaaga 1500
gaacgtgaaa tggacacagt gttagaagat gaagcttgca gaaaacatat aagacaaatg 1560
attcaatctt tatgggttga actaaacggg gagctagtgg cgtcaagtgc gcttccctta 1620
agtatcatca aggcagcttt taacctaagt agaaccgcac aagttattta ccagcacggc 1680
gatgataaca aaacctcctc agttgaagac cacgttcaag ctctattatt ccgtccagtt 1740
tcaagtaacg gacatgccca aattacaatg cat 1773
<210> 101
<211> 1773
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 101
atgtcgcgat tcgttaccat gccctcccac gtgcttccat cgtccttcgt tgctccatct 60
ctccaggttt cgtcgtctcc ctgttcttgg agaactcgac cttccccctg tacttcttgt 120
catctgtccc ccagctcgtc gtccaaacct ctgctgggta gccacgatta cagcctcctg 180
aagtctctca ccctttctcc tcacgctgtt aactccgagg ccgattcctc cacccgacgg 240
atgaaagagg tgaaggagcg aacttgggaa gccttctacc gagcttggga ctcccgggcc 300
gcaatggaga tggtcgacac tgtcgaaaga ctgggtctgt cttaccactt tgaggatgag 360
atcaacgctc tcctccagag attctgtgat tggaacgcct cggaggacct gttcacgacc 420
gctctccgat tccgactcct gagacagaac ggcttcccca cacattcgga cgtcttcggc 480
aagttcatgg acaagaacgg aaagttcaag gagtcgctca cggaagacat ctggggaatg 540
ctttctctcc atgaggcctc gcacctgggc gctaagaatg aggaggtcct cgccgaggcc 600
aaggagttta cccgaatcca ccttatccag agcatgcctc acatggagcc tcacttttcc 660
tcccacgtcg gtcgagccct tgagctgccc cgacatcttc gaatggtccg actggaggct 720
cgtaactaca ttggcgagta ctctagagag tccaacccca acctggcctt cctcgaactc 780
gccaagctgg atttcgacat ggtccagtcg ctgcaccaga aggagctcgc cgagattgtc 840
cgatggtgga agcaactcgg ccttgtcgac aaactggatt tcgctcgaga tcgacccatg 900
gagtgcttcc tgtggaccgt tggtattttc cccgaccccc gtcactcttc ctgccgaatc 960
gagctgacca aggccatcgc cattctgctc gttattgatg acatttacga ctcctacgga 1020
agccttgacg agctcgcact cttcaccgac gccgtgaagc gatgggatct gggtgccatg 1080
gatcaactgc ccgagtacat gaagatctgc tacatggccc tttacaacac aaccaacgac 1140
atcgcttacc gaatccttaa ggagcacgga tggtctgtta ttgaggacct caagcgaacg 1200
tggatggata tcttcggagc ctttctggct gaggcctact gtttcaaggg aggccatgtc 1260
ccttcgctcg aagagtatct gaccaacgct gtcaccaccg gtggtactta catggctctg 1320
gtccacgcat tcttcctgat gggtcaagga gtcacaagag agaatatggc tatgcttaag 1380
ccctatccca acattttttc ttgctctgga aagattctcc ggctctggga tgacctcggc 1440
actgctcgag aggagcagga gcgaggcgat aacgcttcct cgattgagtg ctacaagcga 1500
gagcgagaga tggataccgt ccttgaggac gaggcttgcc gtaagcatat tcgtcaaatg 1560
attcagtcgc tctgggttga gctgaacgga gaactggttg catcgtccgc cctccctctg 1620
tcgatcatca aagccgcttt caacctctcg agaactgctc aggtgattta ccagcacgga 1680
gatgacaaca agaccagctc tgtcgaagac cacgtccagg ctctgctttt cagacctgtg 1740
tcctcgaacg gtcacgctca aattaccatg cac 1773
<210> 102
<211> 501
<212> PRT
<213> 赤豆（Phaseolus angularis）（红豆（Azuki bean））（赤豆（Vigna angularis））
<400> 102
Met Glu Thr Asn Ser Thr Leu Phe Phe Thr Phe Thr Val Leu Leu Ile
1 5 10 15
Thr Ile Ile Thr Phe Leu Lys Ala Leu Lys Ser Leu Phe Thr Pro Asn
20 25 30
Lys Ser Lys Ser Asn Leu Pro Pro Gly Pro Lys Gly Leu Pro Leu Val
35 40 45
Gly Asn Leu Leu Gln Leu Gly Ala Lys Pro His Gln Thr Leu Ala Thr
50 55 60
Leu Ala Asn Ile His Gly Ser Ile Met Ser Leu Lys Leu Gly Gln Glu
65 70 75 80
Thr Thr Ile Val Met Ser Ser Ala Glu Ala Ala Lys Gly Val Leu Gln
85 90 95
Ile His Asp His Phe Leu Ser Asn Arg Lys Ile Pro Asp Ala Met Arg
100 105 110
Gly Ser Ser His Asp His Phe Ser Leu Pro Phe Met Pro Val Ser Gln
115 120 125
Gln Trp Arg Glu Leu Arg Lys Leu Cys Asn Glu Leu Leu Phe Ser Asn
130 135 140
Lys Asn Leu Asp Ala Thr Gln Gly Leu Arg Ser Lys Lys Val Arg Glu
145 150 155 160
Leu Tyr Ser Asp Ile His Arg Ser Ser Leu Lys Gly Glu Pro Val Asn
165 170 175
Ile Gly Arg Leu Ala Phe Lys Thr Thr Ile Asn Gln Leu Ser Asn Thr
180 185 190
Ile Tyr Ser Glu Asp Phe Leu Gln Ser Ala Glu Lys Ala Gly Glu Met
195 200 205
Lys Glu Leu Val Thr Asn Ile Met Lys Glu Val Gly Arg Pro Asn Leu
210 215 220
Ala Asp Cys Phe Pro Val Leu Lys Met Ile Asp Pro His Gly Ile Arg
225 230 235 240
Arg Arg Thr Gly Ser Tyr Phe Ser Lys Leu Leu Asn Ile Phe Lys Ser
245 250 255
Leu Ile His Lys Arg Leu Glu Leu Arg Lys Asp Ala Ala Gly Tyr Cys
260 265 270
Thr Lys Lys Asp Met Leu Asp Ala Met Leu Asn Asp Ala Gln His Lys
275 280 285
Met Asp Ile Val Lys Ile Gln Arg Leu Ser Leu Asp Leu Phe Val Ala
290 295 300
Gly Thr Asp Thr Val Thr Ser Thr Val Glu Trp Ala Met Ala Glu Leu
305 310 315 320
Leu His Asn Pro His Val Met Ser Lys Ala Lys Glu Glu Leu Glu Arg
325 330 335
Ile Ile Gly Lys Asp Asn Leu Val Glu Glu Ser Asp Ile Ala Lys Leu
340 345 350
Pro Tyr Leu Gln Ala Ile Val Lys Glu Thr Phe Arg Leu His Pro Ala
355 360 365
Val Pro Leu Leu Leu Pro Arg Lys Ala Glu Val Glu Phe Glu Met His
370 375 380
Gly Tyr Thr Ile Pro Lys Gly Ala Gln Val Leu Ile Asn Val Trp Ala
385 390 395 400
Ile Gly Arg Asp Pro Asn Leu Trp Glu Lys Pro Arg Leu Phe Trp Pro
405 410 415
Glu Arg Phe Leu Glu Ser Glu Ile Asp Phe Lys Gly Arg Ser Phe Glu
420 425 430
Leu Thr Pro Phe Gly Gly Gly Arg Arg Ile Cys Pro Gly Leu Pro Leu
435 440 445
Ala Ile Arg Leu Val Phe Leu Met Leu Gly Leu Phe Ile Asn Ser Phe
450 455 460
Asp Trp Glu Leu Gln Asp Ile Gln Pro Glu Asp Met Asn Met Asp Glu
465 470 475 480
Asn Phe Gly Leu Thr Leu Glu Lys Ala Gln Pro Val Leu Ala Ile Pro
485 490 495
Ile Ile Pro Lys His
500
<210> 103
<211> 1503
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 103
atggagacta actctaccct tttctttaca ttcaccgtcc tcctgatcac aatcattacg 60
ttcctgaagg cgttgaaatc tttgtttacc cctaacaagt caaaatctaa cctcccaccc 120
ggtcccaagg gccttccttt ggttggcaac ctgcttcagt tgggcgcgaa gccgcaccag 180
accctcgcaa ccctcgcaaa catccacgga tccatcatgt ccctcaaatt gggtcaagag 240
acaaccatcg tgatgtcttc cgctgaggcc gcaaagggag ttctgcagat tcatgatcat 300
ttcctctcaa accgcaaaat ccccgacgca atgcgcggct cctcccacga ccacttctcc 360
cttcctttca tgcctgtttc ccaacagtgg cgagaattgc gaaagctttg caacgaactc 420
ctgttctcca acaagaacct ggacgctaca caaggcctgc gctccaaaaa ggttcgagaa 480
ctgtacagcg atattcaccg cagctcgctg aagggcgagc ccgtgaacat cggtcgtctg 540
gcttttaaaa cgaccatcaa ccaactttct aacaccatct actcggaaga cttcttgcaa 600
tccgccgaga aagcgggcga aatgaaggaa ctcgttacca atattatgaa ggaggttggc 660
cgcccgaacc ttgccgactg cttccccgtt ctgaaaatga tcgaccccca cggcatccgc 720
cgtcgaaccg gctcttactt cagcaagctg cttaatatct tcaaatccct gatccacaag 780
cgtctggaac ttcgtaaaga tgctgcgggc tactgtacta aaaaggatat gcttgacgcg 840
atgctgaacg acgcccaaca caaaatggac atcgttaaga tccagcgtct ctctcttgat 900
ttgttcgttg caggaacgga taccgtcacc tccactgtcg agtgggctat ggctgaactg 960
cttcacaacc cccacgttat gtcaaaggct aaagaggaac tcgaacgcat tatcggtaag 1020
gataatctgg tggaggagtc agacatcgcc aagttgcctt acttgcaggc tatcgtgaag 1080
gagaccttcc gtctccaccc agcagtcccc cttttgctgc cacgcaaagc cgaggttgaa 1140
tttgaaatgc acggttacac aatccccaaa ggcgcccagg tcctgatcaa tgtctgggcc 1200
attggtcgcg acccgaactt gtgggaaaag ccacgcctgt tttggcccga gcgatttctt 1260
gaatccgaga tcgacttcaa gggccgatcg ttcgagctca ccccgttcgg cggcggccgt 1320
cgcatctgcc ctggactccc attggcgatc cgacttgttt ttctgatgct cggcctgttt 1380
attaactctt tcgattggga acttcaggac attcagccgg aagacatgaa tatggacgaa 1440
aacttcggtc tgactcttga aaaggcccag ccagttctcg caattccaat cattccaaaa 1500
cac 1503
<210> 104
<211> 1503
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 104
atggaaacca acagtacttt attctttaca tttacagtcc ttcttatcac tatcataaca 60
tttctcaagg cgctcaaaag tttattcacg ccgaataaat caaaatcaaa tcttccgcct 120
ggaccaaaag gactcccgct ggttggaaat cttttgcaac tcggggcaaa acctcatcag 180
acattagcta cgcttgctaa cattcacgga agcattatgt ctctgaagtt aggtcaagaa 240
acaacaattg tcatgtcttc ggcggaggcg gctaaaggcg ttttgcagat ccacgatcat 300
tttctgagca accggaaaat tcctgacgcc atgcgtggta gctcacatga tcatttctca 360
ttaccgttca tgccagtaag ccaacaatgg cgcgagctga gaaaactgtg taatgagtta 420
ttattttcaa ataaaaactt ggatgcaacg caggggctga gaagcaaaaa agttcgtgaa 480
ctgtattcag atatacatcg ctcgtctctt aaaggcgaac cggtcaacat cgggagactt 540
gcttttaaaa caacaattaa ccaattgagc aatacgatct actcggagga cttccttcaa 600
agcgcggaaa aagcaggtga aatgaaggaa ctggtcacaa acatcatgaa ggaagtggga 660
cgtccgaatc tggctgactg ttttccggtg ctgaaaatga ttgatcctca cggaattcgc 720
cgccggacag gttcatattt ctcaaaactt ctgaatatat tcaaaagctt aattcataaa 780
cgcttagagt tgcgcaaaga tgctgcaggc tattgcacga agaaagatat gctggatgcg 840
atgctgaacg atgctcaaca taaaatggat atcgtgaaaa tccagcggct ctcactcgac 900
ttatttgttg ctggcacaga taccgtcacc tcgaccgtcg agtgggccat ggcagagttg 960
ctgcataatc cacacgtaat gtccaaagct aaagaagaac tggaaagaat cattggcaag 1020
gataacctcg ttgaagaaag cgacatcgcg aaacttcctt acttacaggc gattgtaaag 1080
gagacgttta gactgcatcc agcagtacct ctgcttcttc cgcgcaaagc tgaagtagaa 1140
tttgaaatgc atggatatac gatcccgaaa ggggcacagg tactcatcaa tgtgtgggcg 1200
attggcagag atccaaattt gtgggaaaaa cctcgtttgt tttggcctga aagattcctg 1260
gagtcagaaa ttgacttcaa gggcagatcc ttcgaactca cgccgtttgg cgggggccgg 1320
cgtatttgcc ctggcctgcc gttagccatt agattggttt tcttgatgtt aggcttattc 1380
atcaatagct tcgattggga gctccaggat attcaaccgg aagacatgaa tatggacgaa 1440
aactttggct taacactgga gaaagcacag ccggtactgg ccattccgat catcccgaaa 1500
cat 1503
<210> 105
<211> 1503
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 105
atggaaacta acagcacttt attttttaca ttcacagtct tacttattac gataattacg 60
ttcctgaaag ctctgaaatc actatttact ccaaataaaa gtaagtctaa cttgccccct 120
ggcccaaagg gtttgccact tgttggaaat ctattgcaat taggtgcaaa acctcatcaa 180
acactggcta ctttggctaa tatacatggg tccattatga gcctaaagtt gggccaagag 240
acaactatag tgatgtcttc tgctgaagca gccaagggtg tcttgcaaat acatgatcat 300
ttcctgagca atagaaagat tcctgacgct atgaggggat caagccacga tcatttctca 360
ttgccattca tgcctgtatc ccagcaatgg agagaattaa gaaaattatg taatgaactg 420
ttattttcta acaagaatct ggatgcaaca cagggcttaa gatcaaagaa agttagggaa 480
ttatattcag atatccatcg tagttcttta aaaggcgaac cagtcaacat cggcagactg 540
gcatttaaaa ccaccattaa ccagttgtct aataccattt atagcgaaga tttcttacaa 600
tctgccgaaa aagctggcga gatgaaagaa ttggttacta atatcatgaa ggaagttggt 660
aggccgaacc ttgcagactg ctttcctgtc ttgaagatga tagatccaca tggcattaga 720
agaagaacgg gttcttattt ttctaagttg cttaatattt tcaaatcatt gatccacaag 780
aggttggagt tgagaaaaga tgcggcagga tattgtacga aaaaagacat gctggatgca 840
atgcttaacg atgctcagca taagatggat atagttaaga tccaaagatt atctttggat 900
ttgttcgtcg ccggtactga tactgttacc tccactgtag aatgggccat ggcagaattg 960
cttcataacc cgcatgtcat gtcaaaagct aaagaagaat tggaaagaat tattggcaag 1020
gataatctgg tagaggaaag tgacatagct aaacttccat acttgcaagc cattgttaag 1080
gagactttta gattgcatcc agcagtgcct cttcttttgc caagaaaggc tgaagtggaa 1140
tttgaaatgc acggctacac aattccaaaa ggagctcaag tgttgatcaa tgtatgggcc 1200
attggtcgtg acccaaatct atgggaaaaa ccacgtttat tctggccaga gagatttttg 1260
gaatcagaaa ttgacttcaa gggcaggtca ttcgagttga ccccattcgg cggtggtaga 1320
aggatatgtc ctggcttacc acttgccatc cgtttagtat ttttgatgct tgggctattc 1380
attaactctt ttgactggga gcttcaagat attcagcccg aggatatgaa catggatgag 1440
aatttcgggc tgacacttga aaaagcgcaa cctgtattag ctattccgat aattccaaaa 1500
cat 1503
<210> 106
<211> 1503
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 106
atggagacca actcgactct gttctttact tttactgttc tcctgatcac cattatcacc 60
ttcctcaagg cactgaagtc gctgttcacc cccaataagt cgaagagcaa cctccccccc 120
ggtcctaagg gcctccctct ggtgggaaac ctcctccagc tcggcgccaa acctcatcag 180
actctggcta ccctggccaa cattcacggc agcatcatgt cgctcaagct tggtcaggag 240
actactatcg tcatgtcgtc ggccgaggct gctaagggcg tccttcagat ccacgatcac 300
ttcctgtcta accgaaagat ccccgacgct atgagaggat cttctcacga ccatttctct 360
ctccctttca tgccagtctc gcagcagtgg cgagagcttc gaaagctctg taacgagctt 420
ctcttctcta ataagaacct tgatgctact cagggtctca gatctaagaa ggttcgagag 480
ctgtattcgg acattcatcg atcttctctg aagggtgagc ccgtcaacat tggtagactg 540
gctttcaaga caacaattaa tcagctctct aacacaattt acagcgagga cttcctccag 600
tcggctgaga aggccggtga aatgaaggaa ctcgtcacca acatcatgaa ggaagtgggc 660
cggcccaacc ttgctgattg cttccccgtt ctcaagatga ttgaccccca tggaatccgg 720
cgacggaccg gctcctattt ttccaagctc ctgaacattt tcaagtccct catccacaag 780
cgactcgaac tccgaaaaga cgccgccggt tactgtacca aaaaagacat gcttgacgct 840
atgctcaacg acgcacagca caagatggat attgtcaaga ttcagcgact gtccctcgac 900
ctgttcgtgg ctggaaccga caccgttaca tctacagtcg aatgggctat ggccgaactg 960
ctgcataacc ctcacgtgat gagcaaggct aaggaggagc ttgagcgaat catcggaaag 1020
gacaacctcg tggaggagtc ggacatcgcc aagctgcctt accttcaagc cattgttaag 1080
gagacctttc gactgcaccc agctgttcca ctgctgctcc caagaaaagc tgaggtcgag 1140
ttcgaaatgc acggatatac aatccccaag ggagctcagg ttcttatcaa cgtctgggct 1200
attggacgag accccaacct ctgggagaag cctcgactct tttggcctga gcgattcctc 1260
gagagcgaga ttgactttaa gggtagatct ttcgaactga ccccctttgg tggcggacga 1320
cggatctgcc ccggtctgcc cctcgctatt cgactggttt tcctcatgct tggcctcttc 1380
atcaatagct ttgactggga gcttcaagac attcagcctg aagacatgaa catggacgag 1440
aacttcggcc tgaccctgga gaaggcccag cccgtcctgg caatccccat catccccaag 1500
cac 1503
<210> 107
<211> 495
<212> PRT
<213> 川西獐牙菜（Swertia mussotii）（龙胆科植物（Felwort））
<400> 107
Met Asp Phe Asp Phe Leu Thr Ile Ala Ile Gly Phe Leu Phe Thr Ile
1 5 10 15
Thr Leu Tyr Gln Ala Leu Asn Phe Phe Ser Arg Lys Ser Lys Asn Leu
20 25 30
Pro Pro Gly Pro Ser Pro Leu Pro Leu Ile Gly Asn Leu His Leu Leu
35 40 45
Gly Asp Gln Pro His Lys Ser Leu Ala Lys Leu Ala Lys Lys His Gly
50 55 60
Pro Ile Met Gly Leu Gln Leu Gly Gln Val Thr Thr Ile Val Val Thr
65 70 75 80
Ser Ser Gly Met Ala Lys Glu Val Leu Gln Lys Gln Asp Leu Ala Phe
85 90 95
Ser Ser Arg Ser Ile Pro Asn Ala Ile His Ala His Asp Gln Tyr Lys
100 105 110
Tyr Ser Val Ile Trp Leu Pro Val Ala Ser Arg Trp Arg Gly Leu Arg
115 120 125
Lys Ala Leu Asn Ser Asn Met Phe Ser Gly Asn Arg Leu Asp Ala Asn
130 135 140
Gln His Leu Arg Ser Arg Lys Val Gln Glu Leu Ile Ala Tyr Cys Arg
145 150 155 160
Lys Ser Ser Gln Thr Gly Asp Ala Ile Asp Val Gly Arg Ala Ala Phe
165 170 175
Arg Thr Ser Leu Asn Leu Leu Ser Asn Thr Met Phe Ser Lys Asp Leu
180 185 190
Thr Asp Pro Tyr Ser Asp Ser Ala Lys Glu Phe Lys Asp Leu Val Trp
195 200 205
Asn Val Met Val Glu Ala Gly Lys Pro Asn Leu Val Asp Tyr Phe Pro
210 215 220
Leu Leu Asp Lys Val Asp Pro Gln Gly Ile Arg Lys Arg Met Thr Ile
225 230 235 240
His Phe Gly Lys Ile Leu Glu Leu Phe Gly Gly Leu Ile Asp Glu Arg
245 250 255
Leu Gln Gln Lys Lys Ala Lys Gly Val Asn Asp Asp Val Leu Asp Val
260 265 270
Leu Leu Thr Thr Ser Glu Glu Ser Pro Glu Glu Ile Asp Arg Thr His
275 280 285
Ile Gln Arg Met Cys Leu Asp Leu Phe Val Ala Gly Thr Asp Thr Thr
290 295 300
Ser Ser Thr Leu Glu Trp Ala Met Ser Glu Met Leu Lys Asn Pro Glu
305 310 315 320
Lys Met Lys Ala Ala Gln Ala Glu Leu Ala Gln Val Ile Gly Lys Gly
325 330 335
Lys Ala Val Glu Glu Ala Asp Leu Ala Arg Leu Pro Tyr Leu Arg Cys
340 345 350
Ala Ile Lys Glu Thr Leu Arg Ile His Pro Pro Val Pro Leu Leu Ile
355 360 365
Pro Arg Arg Thr Glu Gln Glu Val Glu Val Cys Gly Tyr Thr Val Pro
370 375 380
Lys Asn Ser Gln Val Leu Val Asn Val Trp Ala Ile Ser Arg Asp Asp
385 390 395 400
Ala Ile Trp Lys Asp Pro Leu Ser Phe Lys Pro Glu Arg Phe Leu Glu
405 410 415
Ser Glu Leu Glu Met Arg Gly Lys Asp Phe Glu Leu Ile Pro Phe Gly
420 425 430
Ala Gly Arg Arg Ile Cys Pro Gly Leu Pro Leu Ala Val Arg Met Val
435 440 445
Pro Val Met Leu Gly Ser Leu Leu Asn Ser Phe Asp Trp Lys Leu Glu
450 455 460
Gly Gly Ile Ala Pro Lys Asp Leu Asp Met Glu Glu Lys Phe Gly Ile
465 470 475 480
Thr Leu Gln Lys Ala His Pro Leu Arg Ala Val Ala Thr Pro Leu
485 490 495
<210> 108
<211> 1485
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 108
atggatttcg atttccttac gattgccatt ggatttcttt ttacaattac cttgtatcag 60
gctctgaatt ttttcagtcg taaatcgaaa aaccttccgc caggaccatc accgctgcct 120
ttgataggga atcttcattt gttaggtgac cagcctcata aatcgttggc caagcttgcg 180
aagaaacatg gtcctattat gggcttacaa ttgggccaag tgacgactat tgttgttacg 240
tctagcggca tggccaagga agttcttcaa aagcaggatt tagctttttc atcccgttct 300
attccaaatg cgatacatgc acatgatcag tacaagtact cagtgatttg gcttcctgtg 360
gcatcacggt ggcgtgggct gcgcaaagct ttgaattcca acatgttttc cggaaacaga 420
ctggatgcga accaacatct gcggtctcgc aaagtgcagg aattaattgc ctattgcaga 480
aaatcaagcc aaactggtga cgcaattgac gtagggcgag ccgcctttcg aacatctctt 540
aacctcctgt ctaacacgat gttttctaaa gacctcactg atccttactc cgattccgcg 600
aaagaattca aggatttggt ctggaacgtg atggttgagg cgggaaaacc taacttggtg 660
gattattttc cgctccttga taaagtggac ccgcaaggaa ttcggaagcg catgacgatc 720
cattttggta aaattctcga gctttttggc ggattaattg atgaacgtct tcagcaaaaa 780
aaggcgaagg gagtgaacga cgacgttctt gacgtgcttc tgacgacctc tgaggaatcc 840
ccagaagaga ttgatcggac tcatattcag cgtatgtgcc tggatttgtt tgtggccggt 900
acggacacta cctcctccac gctcgaatgg gcaatgtcgg aaatgttaaa aaatcctgag 960
aaaatgaaag cggctcaagc agaactggcc caagtcatcg gcaaaggcaa agccgtagaa 1020
gaagccgatc ttgctcgctt gccgtattta cggtgcgcaa ttaaagaaac cctgagaatc 1080
catccgcctg tcccgctttt aatccctcga agaacagaac aagaggtcga agtctgtgga 1140
tataccgtgc ctaaaaattc acaagtgctc gtaaacgtct gggcgattag tcgtgacgac 1200
gcgatatgga aagacccgct tagttttaaa ccggaacggt tcctcgagag cgagttagaa 1260
atgcggggta aggatttcga gctgattcct ttcggggcgg gtagaagaat ctgccctggc 1320
ctgccgctgg cggtgcggat ggttccagtt atgcttggtt ctctgcttaa ttcattcgat 1380
tggaaactgg agggtggaat cgcgccgaag gacctggata tggaggagaa atttggaatt 1440
actttacaga aggcgcatcc tcttcgcgca gttgccacgc ctctc 1485
<210> 109
<211> 1485
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 109
atggattttg attttttaac cattgcaatt ggttttttat tcaccattac gttgtatcaa 60
gcgttaaatt ttttttctag gaagtccaaa aaccttccgc caggtccttc tccactgcca 120
ctaatcggca acctacatct tttgggtgat cagcctcaca aatcattggc aaaattggcg 180
aagaaacacg gcccgattat gggtctacaa ctgggtcaag taacgaccat agttgtaaca 240
tctagtggca tggctaaaga agtacttcaa aagcaagatt tagctttttc ctcaagatct 300
ataccaaacg caatacatgc ccatgatcaa tacaagtaca gtgttatatg gctgccagtt 360
gcttccagat ggcgtgggct acgtaaagcg ctaaattcta acatgttctc tggcaataga 420
ttggacgcca accagcattt aagatctaga aaagtgcaag aattaattgc atactgtaga 480
aaatctagtc aaacagggga tgccatagat gtagggcgtg ctgcgtttag aaccagcttg 540
aatttattgt ctaacactat gttttcaaaa gacttgaccg atccctacag cgactccgca 600
aaggaattta aagatttagt ttggaacgtt atggtagagg ctggaaaacc gaatttggtg 660
gattattttc ccttgttgga taaggtcgac ccccagggta ttagaaagag aatgacaatt 720
cacttcggaa agatattgga actgtttggt ggattgatag acgaacgttt acaacaaaag 780
aaagccaaag gtgtgaatga tgatgtgctt gatgttttgc taactacaag cgaggaaagt 840
ccggaagaaa tagataggac ccatatccaa aggatgtgtt tggacttgtt tgttgccgga 900
actgatacca ctagttcaac attagaatgg gctatgtctg agatgttgaa aaatcctgag 960
aaaatgaaag ctgcacaggc agagttggcg caagtgatag gtaaaggtaa agctgtagag 1020
gaagcggatt tggctagatt accctatttg aggtgtgcta tcaaggaaac tcttaggatt 1080
catcctccag taccactatt aattccacgt agaaccgaac aggaagtgga agtgtgtgga 1140
tataccgtac cgaagaactc tcaagtgctg gtaaatgtgt gggccatttc tagagatgat 1200
gctatatgga aagacccatt atcttttaaa cccgagaggt ttctggaaag cgagttggaa 1260
atgagaggca aagactttga attgatccca tttggggcgg ggagaagaat ctgtcctgga 1320
ttgccgttgg cagtgagaat ggttccagtt atgctgggtt ctctgttgaa cagtttcgat 1380
tggaagttag agggagggat agcccctaaa gatctggaca tggaagaaaa gtttggcatt 1440
actcttcaaa aagcccatcc actaagggcc gtagctacac cgctg 1485
<210> 110
<211> 1485
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 110
atggatttcg atttcctgac cattgcaatc ggctttcttt ttaccattac tctttaccag 60
gctcttaact tcttctctcg aaagtctaag aacctccctc ctggaccttc ccccctgccc 120
ctgatcggaa accttcacct cctcggcgac cagccccaca agtccctggc taagcttgct 180
aaaaagcacg gccccatcat gggtcttcag ctgggacagg ttacaactat tgttgttacc 240
agctccggaa tggctaagga ggtgctccag aaacaggacc ttgccttctc ctcgagatcc 300
atccccaacg ccatccatgc ccacgatcag tacaagtact ctgttatttg gctccctgtg 360
gcctctcgtt ggcgaggtct ccgaaaagct ctgaattcca acatgttcag cggtaaccgt 420
ctggacgcaa atcagcatct ccggtctaga aaggttcagg agctgattgc ttactgtcga 480
aagtcttcgc agacaggtga cgctatcgat gtgggtagag cagccttccg aacttcgctg 540
aacctcctct cgaataccat gttctctaag gatcttaccg atccatacag cgattctgcc 600
aaggagttta aggacctggt ttggaacgtc atggtcgagg caggtaagcc taacctggtt 660
gattactttc ccctgcttga taaagtggac cctcagggaa ttcgaaagag aatgaccatt 720
cattttggta agatcctcga gctgttcggt ggccttatcg acgagcgact ccagcagaaa 780
aaggccaagg gtgttaacga cgacgttctc gacgtgctgc tgacgacatc tgaggagtcc 840
cccgaggaaa ttgaccgaac ccatatccaa agaatgtgtc tggacctctt tgtcgctggt 900
actgacacca cttcctccac actggagtgg gcaatgtcgg aaatgctcaa gaaccctgag 960
aagatgaagg ctgcccaggc cgagcttgcc caggtcatcg gcaagggtaa ggccgtggaa 1020
gaggccgacc tcgctcgact gccctacctt cgatgtgcta tcaaggagac gctgagaatc 1080
cacccccctg tgcccctcct catcccccgt cgaaccgagc aggaagttga ggtctgtggc 1140
tacaccgtcc caaagaactc ccaggttctc gttaacgtgt gggctatttc cagagatgac 1200
gccatctgga aggaccctct gtcttttaag cctgagcgat tcctggagtc cgagctggag 1260
atgcggggaa aggactttga actgatcccc tttggagctg gacgacgaat ctgccccggt 1320
ctccccctgg ccgtccgaat ggtccccgtc atgcttggat cgctcctcaa ctcgtttgac 1380
tggaagcttg aaggcggaat tgctcccaag gaccttgata tggaagagaa gttcggtatc 1440
actctgcaga aagcacaccc cctccgggca gtcgccactc ctctg 1485
<210> 111
<211> 864
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 111
atgacagcag ataacaattc catgcctcac ggtgctgtgt cttcatacgc gaaactggtg 60
cagaaccaaa cacctgagga tattctcgag gaattccccg aaatcatccc tctccagcag 120
cgcccgaaca cccgctcctc tgaaacctct aatgacgaat ccggcgagac atgtttctcc 180
ggccacgacg aagagcaaat taagctgatg aacgaaaatt gcatcgtgtt ggactgggat 240
gacaacgcca ttggagccgg taccaaaaag gtgtgccatc tgatggagaa cattgaaaag 300
ggactgctcc atcgagcttt ctcagtgttc attttcaatg aacagggtga gctcctgctg 360
caacagcgcg caacggaaaa gattaccttc cccgacctct ggactaacac ctgctgctca 420
cacccattgt gtatcgatga cgaacttggc ttgaagggca agcttgatga taagattaag 480
ggcgcaatca cggccgctgt tcgtaaactc gatcacgaac ttggaattcc tgaagacgaa 540
accaagactc gcggcaaatt tcacttcttg aatcgtatcc actatatggc gccttccaac 600
gagccttggg gcgagcacga gatcgactat attttgttct acaagatcaa tgcaaaggag 660
aacctgactg tgaacccgaa tgtgaatgaa gtgcgtgact tcaagtgggt ctcaccgaac 720
gatctgaaga ccatgtttgc cgatccatcc tacaaattca ccccgtggtt caaaattatc 780
tgtgaaaact atctgttcaa ttggtgggaa caactcgacg atttgtccga ggtcgagaac 840
gatcgtcaaa tccaccgtat gctt 864
<210> 112
<211> 1485
<212> DNA
<213> 人工
<220>
<223> 突变和/或密码子优化序列
<400> 112
atggattttg atttcttgac aatcgcaatt ggtttcctgt tcaccattac cttgtaccaa 60
gcgctcaact tctttagccg caaatccaag aacctcccac caggtccgag ccccctcccg 120
cttattggca acctgcactt gctgggagat cagccgcaca aatcattggc gaagctcgct 180
aagaagcacg gtccaattat gggtctgcag ttgggccaag ttacaactat cgtcgtgacc 240
tcgtccggaa tggcaaagga ggttttgcaa aaacaagatt tggctttctc gtcccgctcc 300
atccccaacg ctatccacgc acacgaccag tacaagtaca gcgtgatttg gctgcccgtt 360
gctagccgct ggcgcggatt gcgcaaggca ctgaattcca acatgttctc tggcaaccgc 420
ttggacgcca accaacacct tcgatcccgc aaagtgcaag aactgatcgc ttactgtcgc 480
aaaagctctc agaccggcga tgccattgac gtaggccgcg cggcatttcg aacctcgctc 540
aaccttctct cgaacacgat gttttcaaaa gaccttaccg acccatattc agattccgcc 600
aaggagttca aggaccttgt ttggaacgtg atggtcgaag ccggcaagcc gaatcttgtc 660
gattactttc cgttgctcga taaggtggat ccccagggaa tccgcaagcg tatgaccatc 720
cacttcggaa aaatcctgga gctgttcggt ggcctgatcg acgagcgcct tcaacagaag 780
aaggccaagg gagttaatga cgacgtgctg gatgtgcttc tcaccacctc tgaagagtcc 840
cctgaggaga tcgatcgtac acacatccag cgcatgtgtc ttgatttgtt cgtggctgga 900
acagatacca caagctcaac cttggagtgg gcaatgtcgg agatgcttaa gaacccggaa 960
aagatgaagg ctgcacaggc ggaactggcc caggtaatcg gcaagggtaa ggcagtagaa 1020
gaggctgacc tcgcacgact cccatatctc cgttgtgcta tcaaggagac cctgcgtatt 1080
cacccgccgg tccctcttct tattcctcgc cgcaccgagc aggaggttga agtgtgtggt 1140
tacactgtgc ccaaaaactc ccaagtgctg gtcaacgttt gggcgatctc gcgcgatgac 1200
gccatctgga aagacccttt gtccttcaag ccagagcgtt ttttggaatc cgagctggag 1260
atgcgtggaa aggattttga attgatcccg ttcggtgccg gtcgtcgcat ctgcccgggt 1320
cttccccttg cagtacgcat ggtcccggta atgctgggct cgctcctgaa ctcttttgat 1380
tggaaacttg agggcggtat cgctccaaag gacttggata tggaagaaaa gttcggaatt 1440
actttgcaaa aagcacaccc gttgcgcgca gtcgcaaccc ctctg 1485

Claims (44)

1.一种经改造的微生物细胞，其中所述经改造的微生物细胞表达：
(a)非天然的香叶基二磷酸二磷酸酶(香叶醇合酶)；和
(b)非天然的香叶醇-8羟化酶；
其中所述经改造的微生物细胞产生(6E)-8-羟基香叶醇。
2.权利要求1的经改造的微生物细胞，其中所述经改造的微生物细胞包含一种或多种上游(6E)-8-羟基香叶醇途径酶或上游途径活性调节因子的提高的活性，所述提高的活性为相对于对照细胞的提高。
3.权利要求2的经改造的微生物细胞，其中所述一种或多种上游(6E)-8-羟基香叶醇途径酶选自ATP-柠檬酸合酶、乙酰CoA合酶、硫解酶、羟甲基戊二酰辅酶A合酶(HMG-CoA合酶)、羟甲基戊二酰辅酶A还原酶(HMG-CoA还原酶)、甲羟戊酸激酶、磷酸甲羟戊酸激酶、二磷酸甲羟戊酸脱羧酶、异戊烯基二磷酸δ-异构酶和香叶基二磷酸合酶。
4.权利要求3的经改造的微生物细胞，其中所述一种或多种上游(6E)-8-羟基香叶醇途径酶包括异戊烯基二磷酸δ-异构酶。
5.权利要求1-4中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包含消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶的降低的活性，所述降低的活性为相对于对照细胞的降低。
6.权利要求5的经改造的微生物细胞，其中所述消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶包括双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶和/或香叶基焦磷酸合酶。
7.权利要求1-6中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞还表达反馈去调控的HMG-CoA还原酶。
8.权利要求1-7中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括相对于对照细胞，提高的乙酰CoA可用性，这是由于乙酰CoA的合成速率较高和/或乙酰CoA的降解速率较低。
9.一种经改造的微生物细胞，其中所述经改造的微生物细胞包含：
(a)表达非原生的天然香叶基二磷酸二磷酸酶(香叶醇合成酶)的方式；和
(b)表达非天然的香叶醇-8-羟化酶的方式；
其中所述经改造的微生物细胞产生(6E)-8-羟基香叶醇。
10.权利要求9的经改造的微生物细胞，其中所述经改造的微生物细胞包含提高一种或多种上游(6E)-8-羟基香叶醇途径酶或上游途径活性调节因子的活性的方式。
11.权利要求10的经改造的微生物细胞，其中所述一种或多种上游(6E)-8-羟基香叶醇途径酶选自ATP-柠檬酸合酶、乙酰CoA合酶、硫解酶、羟甲基戊二酰辅酶A合酶(HMG-CoA合酶)、羟甲基戊二酰辅酶A还原酶(HMG-CoA还原酶)、甲羟戊酸激酶、磷酸甲羟戊酸激酶、二磷酸甲羟戊酸脱羧酶、异戊烯基二磷酸δ-异构酶和香叶基二磷酸合酶。
12.权利要求11的经改造的微生物细胞，其中所述一种或多种上游(6E)-8-羟基香叶醇途径酶包括异戊烯基二磷酸δ-异构酶。
13.权利要求9-12中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括降低消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶的活性的方式，所述降低的活性为相对于对照细胞的降低。
14.权利要求13的经改造的微生物细胞，其中所述消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶包括双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶和/或香叶基焦磷酸合酶。
15.权利要求9-14中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞还包含表达反馈去调控的HMG-CoA还原酶的方式。
16.权利要求9-15中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括相对于对照细胞，提高乙酰CoA的可用性的方式，这是由于乙酰CoA的合成速率较高和/或乙酰CoA的降解速率较低。
17.权利要求1-16中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括真菌细胞。
18.权利要求17的经改造的微生物细胞，其中所述经改造的微生物细胞包括酵母细胞。
19.权利要求18的经改造的微生物细胞，其中所述酵母细胞为酵母属(Saccharomyces)的细胞。
20.权利要求19的经改造的微生物细胞，其中所述酵母细胞为酿酒酵母(Saccharomyces cerevisiae)种的细胞。
21.权利要求18的经改造的微生物细胞，其中所述酵母细胞为亚罗酵母属(Yarrowia)的细胞。
22.权利要求21的经改造的微生物细胞，其中所述酵母细胞为解脂亚罗酵母(Yarrowialipolytica)种的细胞。
23.权利要求1-22中任一项的经改造的微生物细胞，其中所述非天然的香叶醇合酶包括与来自紫苏(Perilla setoyensis)的香叶醇合酶具有至少70％的氨基酸序列同一性的香叶醇合酶。
24.权利要求1-22中任一项的经改造的微生物细胞，其中所述非天然的香叶醇合酶包括与来自欧洲葡萄(Vitis vinifera)的香叶醇合酶具有至少70％的氨基酸序列同一性的香叶醇合酶。
25.权利要求1-23中任一项的经改造的微生物细胞，其中所述非天然的香叶醇-8-羟化酶包括与来自赤豆(Phaseolus angularis)的香叶醇-8-羟化酶具有至少70％的氨基酸序列同一性的香叶醇-8-羟化酶。
26.权利要求4或12-25中任一项的经改造的微生物细胞，其中所述异戊烯基二磷酸δ-异构酶的提高的活性是通过异源表达异戊烯基二磷酸δ-异构酶实现的。
27.权利要求26的经改造的微生物细胞，其中所述异源的异戊烯基二磷酸δ-异构酶包括与来自酿酒酵母(Saccharomyces cerevisiae)的异戊烯基二磷酸δ-异构酶具有至少70％的氨基酸序列同一性的异戊烯基二磷酸δ-异构酶。
28.权利要求6和14-27中任一项的经改造的微生物细胞，其中所述消耗一种或多种(6E)-8-羟基香叶醇途径前体的一种或多种酶包括双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶。
29.权利要求28的经改造的微生物细胞，其中双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶与来自大肠杆菌(Escherichia coli)的双功能的(2E，6E)-法呢基二磷酸合酶/二甲基烯丙基反式转移酶具有至少70％的氨基酸同一性，并包含氨基酸置换S80F。
30.权利要求7或15-27中任一项的经改造的微生物细胞，其中所述HMG-CoA还原酶是酿酒酵母HMG-CoA还原酶的变体。
31.权利要求1-30中任一项的经改造的微生物细胞，其中，当培养时，所述经改造的微生物细胞产生水平高于100μg/L培养基的(6E)-8-羟基香叶醇。
32.权利要求1-31中任一项的经改造的微生物细胞的培养物。
33.权利要求32的培养物，其中所述底物包括碳源和氮源，所述氮源选自尿素、铵盐、氨及其任意组合。
34.权利要求32-33中任一项的培养物，其中所述经改造的微生物细胞以一定浓度存在，从而使得所述培养物在600nm下的光密度为10-500。
35.权利要求32-34中任一项的培养物，其中所述培养物包含(6E)-8-羟基香叶醇。
36.权利要求32-35中任一项的培养物，其中所述培养物包含水平高于100μg/L培养基的(6E)-8-羟基香叶醇。
37.培养权利要求1-31中任一项的经改造的微生物细胞的方法，所述方法包括在适合产生(6E)-8-羟基香叶醇的条件下培养所述细胞。
38.权利要求37的方法，其中所述方法包括分批补料培养，初始葡萄糖水平范围为1-100g/L，然后是受控的糖补料。
39.权利要求37或权利要求38的方法，其中所述发酵底物包括葡萄糖和选自尿素、铵盐、氨及其任意组合的氮源。
40.权利要求37-39中任一项的方法，其中在培养过程中所述培养物是pH受控的。
41.权利要求37-40中任一项的方法，其中在培养过程中给所述培养物通气。
42.权利要求37-41中任一项的方法，其中所述经改造的微生物细胞以高于100μg/L培养基的水平产生(6E)-8-羟基香叶醇。
43.权利要求37-42中任一项的方法，其中所述方法还包括从培养物中回收(6E)-8-羟基香叶醇。
44.利用经改造以产生(6E)-8-羟基香叶醇的微生物细胞来制备(6E)-8-羟基香叶醇的方法，所述方法包括：
(a)在微生物细胞中表达非天然的香叶基二磷酸二磷酸酶(香叶醇合酶)；
(b)在微生物细胞中表达非天然的香叶醇-8-羟化酶；
(c)在适当的培养基中，使微生物细胞产生(6E)-8-羟基香叶醇的条件下培养微生物细胞，其中所述(6E)-8-羟基香叶醇被释放到培养基中；和
(d)从培养基中分离(6E)-8-羟基香叶醇。
CN201880088768.5A 2017-12-07 2018-12-06 通过发酵产生(6e)-8-羟基香叶醇的经改造的生物合成途径 Pending CN111699247A (zh)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201762596013P 2017-12-07 2017-12-07
US62/596,013 2017-12-07
PCT/US2018/064351 WO2019113387A1 (en) 2017-12-07 2018-12-06 Engineered biosynthetic pathways for production of (6e)-8-hydroxygeraniol by fermentation

Publications (1)

Publication Number Publication Date
CN111699247A true CN111699247A (zh) 2020-09-22

Family

ID=64746661

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201880088768.5A Pending CN111699247A (zh) 2017-12-07 2018-12-06 通过发酵产生(6e)-8-羟基香叶醇的经改造的生物合成途径

Country Status (7)

Country Link
US (1) US10662415B2 (zh)
EP (1) EP3720944A1 (zh)
JP (1) JP2021505154A (zh)
KR (1) KR20200095534A (zh)
CN (1) CN111699247A (zh)
CA (1) CA3084263A1 (zh)
WO (1) WO2019113387A1 (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113234607A (zh) * 2021-05-26 2021-08-10 华中农业大学 利用米曲霉合成香叶醇的方法
CN114621982A (zh) * 2022-03-16 2022-06-14 嘉兴欣贝莱生物科技有限公司 香叶基二磷酸酯的生物合成方法及其在制备大麻类化合物中的应用
CN114717210A (zh) * 2022-04-08 2022-07-08 四川农业大学 一种杨树香叶基香叶醇还原酶及其编码基因与应用

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20200095534A (ko) 2017-12-07 2020-08-10 지머젠 인코포레이티드 발효에 의한 (6e)-8-히드록시제라니올의 제조를 위해 조작된 생합성 경로
JP2021506316A (ja) 2017-12-21 2021-02-22 ザイマージェン インコーポレイテッド ネペタラクトールオキシドレダクターゼ、ネペタラクトールシンターゼ、及びネペタラクトンを産生する能力がある微生物
NL2032683B1 (en) * 2022-07-18 2024-01-26 Sestina Bio Llc Bioproduction of isoprenoids
WO2024059517A1 (en) * 2022-09-14 2024-03-21 Ginkgo Bioworks, Inc. Biosynthesis of oxygenated hydrocarbons
CN117126755B (zh) * 2023-10-24 2024-03-12 深圳先进技术研究院 一种葡萄糖-乙醇共碳源发酵高产香叶醇的酿酒酵母工程菌及其构建方法和应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101509005A (zh) * 2009-03-02 2009-08-19 山东大学 川西獐牙菜香叶醇-10羟化酶基因SmG10H及其应用

Family Cites Families (135)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3278646A (en) 1963-02-01 1966-10-11 Hercules Inc Heat-sealable films comprising polyolefin and a terpene polymer
US3361849A (en) 1963-08-27 1968-01-02 Hercules Inc Blends of isotactic propylene polymer and hydrogenated terpene polymer
JPS52139011A (en) 1976-05-14 1977-11-19 Nissan Chem Ind Ltd Synthesis of linear unsaturated alcohols
ATE203558T1 (de) 1994-05-30 2001-08-15 Hitachi Chemical Co Ltd Schutzfilm für anstrichfilm
US5500282A (en) 1994-07-15 1996-03-19 Mobil Oil Corporation High moisture barrier OPP film containing high crystallinity polypropylene and terpene polymer
JP3151371B2 (ja) 1995-03-10 2001-04-03 麒麟麦酒株式会社 カロチノイド生産量の増量に有用なｄｎａ鎖
US5753507A (en) 1995-09-22 1998-05-19 Novartis Finance Corporation Plant geraniol/nerol 10-hydroxylase and DNA coding therefor
US5744341A (en) 1996-03-29 1998-04-28 University Of Maryland College Park Genes of carotenoid biosynthesis and metabolism and a system for screening for such genes
US20020035058A1 (en) 1996-05-15 2002-03-21 The University Of Sheffield Isopentenyl pyrophosphate isomerase (IPI) and/or prenyl transferase inhibitors
US6806076B1 (en) 1998-04-14 2004-10-19 Kyowa Hakko Kogyo Co., Ltd. Process for producing isoprenoid compounds by microorganisms and a method for screening compounds with antibiotic or weeding activity
TWI250210B (en) 1998-05-06 2006-03-01 Dsm Ip Assets Bv An isolated DNA sequence coding for an enzyme involved in the mevalonate pathway or the pathway from isopentenyl pyrophosphate to farnesyl pyrophosphate
EP1078045A1 (en) 1998-05-13 2001-02-28 University Of Maryland College Park Methods of modifying the production of isopentenyl pyrophosphate, dimethylallyl pyrophosphate and/or isoprenoids
WO2000001686A1 (en) 1998-07-06 2000-01-13 Eastman Chemical Company Method of producing vitamin e
EP2305825B1 (en) 1998-07-06 2015-01-14 DCV Inc. doing business as Bio-Technical Resourses Method of vitamin production
EP1095002A4 (en) 1998-07-06 2005-08-03 Dcv Inc PROCESS FOR PRODUCING VITAMINS
IL141888A0 (en) 1998-09-22 2002-03-10 Jomaa Pharmaka Gmbh Genes of the 1-deoxy-d-xylulose biosynthetic pathway
US20020119546A1 (en) 1998-11-05 2002-08-29 Falco Saverio Carl Squalene synthesis enzymes
US6916972B2 (en) 1998-11-05 2005-07-12 E. I. Du Pont De Nemours And Company Mevalonate synthesis enzymes
US6558915B1 (en) 1998-12-03 2003-05-06 E. I. Du Pont De Nemours And Company Soybean 1-deoxy-D-xylulose 5-phosphae synthase and DNA encoding thereof
AU2163300A (en) 1998-12-04 2000-06-26 E.I. Du Pont De Nemours And Company Plant 1-deoxy-d-xylulose 5-phosphate reductoisomerase
US6387637B1 (en) 1999-01-15 2002-05-14 Syngenta Participations Ag Herbicide target genes and method
GB9901902D0 (en) 1999-01-28 1999-03-17 New Royal Holloway & Bedford Manipulating isoprenoid expression
WO2000078935A1 (en) 1999-06-22 2000-12-28 Smithkline Beecham Corporation Mevalonate pathway genes
CN1900304A (zh) 1999-07-27 2007-01-24 食品工业发展研究所 代谢控制的工程化
BR0014934A (pt) 1999-10-27 2002-06-18 Unilever Nv Usos de um gene que expressa uma hmg-redutase sem retro-inibição e de um gene heterólogo que expressa uma hmg-redutase sem retro-inibição truncada, método de obtenção de sementes, planta, tecido de planta, sementes, método de obtenção de óleo, e, produto alimentìcio
DE10021688A1 (de) 2000-05-05 2001-11-15 Hassan Jomaa Gene des 1-Desoxy-D-xylulose-Biosynthesewegs
WO2002010398A2 (en) 2000-07-31 2002-02-07 Hahn Frederick M Manipulation of genes of the mevalonate and isoprenoid pathways to create novel traits in transgenic organisms
WO2002031120A1 (en) 2000-10-13 2002-04-18 University Of Utah Research Foundation Systems for screening antibacterial and herbicidal compounds
DE10119905A1 (de) 2001-04-23 2002-10-24 Jomaa Pharmaka Gmbh Inaktivierung von Genen des MEP-Wegs
US6989257B2 (en) 2001-06-06 2006-01-24 Dsm Ip Assets B.V. Isoprenoid production
US20040010815A1 (en) 2001-09-26 2004-01-15 Lange B. Markus Identification and characterization of plant genes
US7172886B2 (en) 2001-12-06 2007-02-06 The Regents Of The University Of California Biosynthesis of isopentenyl pyrophosphate
US7192751B2 (en) 2001-12-06 2007-03-20 The Regents Of The University Of California Biosynthesis of amorpha-4,11-diene
US7547793B2 (en) 2003-03-19 2009-06-16 E.I. Du Pont De Nemours And Company Method for making insect repellent composition
EP1490351B1 (en) 2002-04-03 2007-06-06 E.I. Du Pont De Nemours And Company Production of dihydronepetalactone by hydrogenation of nepetalactone
AU2003234790A1 (en) 2002-05-10 2003-11-11 Kyowa Hakko Kogyo Co., Ltd. Process for producing mevalonic acid
CA2501574C (en) 2002-10-04 2013-12-03 E. I. Du Pont De Nemours And Company Process for the biological production of 1,3-propanediol with high yield
CA2514694A1 (en) 2003-02-05 2004-08-19 Basf Aktiengesellschaft Mevalonate kinase as a target for fungicides
ES2323175T3 (es) 2003-06-12 2009-07-08 Dsm Ip Assets B.V. Mevalonato-quinasas resistentes a la retroalimentacion.
WO2005005650A1 (en) 2003-07-08 2005-01-20 Dsm Ip Assets B.V. Improved production of coenzyme q-10
WO2005054224A2 (en) 2003-11-26 2005-06-16 E. I. Du Pont De Nemours And Company Derivatives of dihydronepetalactone and method for preparation
US7704716B2 (en) 2003-12-10 2010-04-27 The Regents Of The University Of Michigan Geraniol synthase, methods of production and uses thereof
US7741070B2 (en) 2003-12-24 2010-06-22 Massachusetts Institute Of Technology Gene targets for enhanced carotenoid production
MXPA06013502A (es) 2004-05-21 2007-03-01 Univ California Metodo para mejorar la produccion de compuestos isoprenoides.
US8828684B2 (en) 2004-07-27 2014-09-09 The Regents Of The University Of California Genetically modified host cells and use of same for producing isoprenoid compounds
CN101044243B (zh) 2004-08-19 2011-12-28 帝斯曼知识产权资产管理有限公司 类异戊二烯的生产方法
US7375239B2 (en) 2004-12-08 2008-05-20 The United States Of America As Represented By The Secretary Of Agriculture Methods of separating ZE-nepetalactone and EZ-nepetalactone from catnip oil
EP1824969B1 (en) 2004-12-14 2011-03-02 DSM IP Assets B.V. Improved mevalonate kinase
BRPI0614990A2 (pt) 2005-08-19 2011-04-26 Univ California células hospedeiras geneticamente modificadas e uso das mesmas para produção de compostos isoprenóides
EP1931770A4 (en) 2005-10-07 2009-11-25 Univ California NUCLEIC ACIDS CODING FOR MODIFIED CYTOCHROM P450 ENZYME AND METHOD OF USE THEREOF
DE102005063191A1 (de) 2005-12-30 2007-07-05 Basf Ag Verfahren zur enzymatischen Herstellung von Citronellal
WO2007093962A2 (en) 2006-02-14 2007-08-23 Firmenich Sa Method for producing terpenes and mep-transformed microorganisms therefore
EP2040555A2 (en) 2006-05-10 2009-04-01 E.I. Du Pont De Nemours And Company Formulated tick and insect repellent compositions
US8114645B2 (en) 2006-05-19 2012-02-14 The Regents Of The University Of California Methods for increasing isoprenoid and isoprenoid precursor production by modulating fatty acid levels
CA2652801C (en) 2006-05-26 2018-05-22 Amyris Biotechnologies, Inc. Apparatus for making bio-organic compounds
MX284139B (es) 2006-05-26 2011-02-18 Amyris Biotechnologies Inc Produccion de isoprenoides.
WO2008008256A2 (en) 2006-07-07 2008-01-17 The Regents Of The University Of California Methods for enhancing production of isoprenoid compounds by host cells
US8257957B2 (en) 2006-09-26 2012-09-04 The Regents Of The University Of California Production of isoprenoids and isoprenoid precursors
US9521844B2 (en) 2006-12-21 2016-12-20 E I Du Pont De Nemours And Company Solvent addition and removal in the hydrogenation of catmint oil
CN101568528B (zh) 2006-12-21 2012-11-14 纳幕尔杜邦公司 荆芥油氢化中溶剂的加入和移除
BRPI0719505A2 (pt) 2006-12-21 2013-12-10 Du Pont "Processos de preparação de óleo de erva dos gatos hidrogenado"
CN101563334B (zh) 2006-12-21 2012-08-22 纳幕尔杜邦公司 通过氢化荆芥内酯制备二氢荆芥内酯
WO2008094546A2 (en) 2007-01-31 2008-08-07 The Regents Of The University Of California Genetically modified host cells for increased p450 activity levels and methods of use thereof
US8158383B2 (en) 2007-03-16 2012-04-17 The Regents Of The University Of California Methods of generating protein variants
WO2008128159A1 (en) 2007-04-16 2008-10-23 Amyris Biotechnologies, Inc. Production of isoprenoids
WO2009005704A1 (en) 2007-07-03 2009-01-08 The Regents Of The University Of California Methods of increasing isoprenoid or isoprenoid precursor production
US8309323B2 (en) 2007-11-13 2012-11-13 Synthetic Genomics, Inc. Dimethyloctane as an advanced biofuel
SG162265A1 (en) 2007-12-13 2010-07-29 Danisco Us Inc Compositions and methods for producing isoprene
WO2009100231A2 (en) 2008-02-06 2009-08-13 The Regents Of The University Of California Short chain volatile isoprene hydrocarbon production using the mevalonic acid pathway in genetically engineered yeast and fungi
WO2009108774A2 (en) 2008-02-28 2009-09-03 The Regents Of The University Of California Use of synthetic scaffolds for the production of biosynthetic pathway products
US20090253174A1 (en) 2008-04-08 2009-10-08 Zach Serber Expression of Heterologous Sequences
SG165884A1 (en) 2008-04-23 2010-11-29 Goodyear Tire & Rubber Isoprene synthase variants for improved microbial production of isoprene
US7632995B2 (en) 2008-05-09 2009-12-15 Monsanto Technology Llc Plants and seeds of hybrid corn variety CH852179
CA2729801A1 (en) 2008-07-02 2010-01-07 Danisco Us Inc. Compositions and methods for producing isoprene free of c5 hydrocarbons under decoupling conditions and/or safe operating ranges
US9909146B2 (en) 2008-07-04 2018-03-06 Scientist Of Fortune S.A. Production of alkenes by enzymatic decarboxylation of 3-hydroxyalkanoic acids
WO2010031077A1 (en) 2008-09-15 2010-03-18 Danisco Us Inc. Increased isoprene production using mevalonate kinase and isoprene synthase
WO2010031076A2 (en) 2008-09-15 2010-03-18 Danisco Us Inc. Conversion of prenyl derivatives to isoprene
EP2334788A1 (en) 2008-09-15 2011-06-22 Danisco US Inc. Increased isoprene production using the archaeal lower mevalonate pathway
WO2010111707A1 (en) 2009-03-27 2010-09-30 Sapphire Energy, Inc. Variant isoprenoid producing enzymes and uses thereof
US20100297749A1 (en) 2009-04-21 2010-11-25 Sapphire Energy, Inc. Methods and systems for biofuel production
WO2010141452A1 (en) 2009-06-01 2010-12-09 Amyris Biotechnologies, Inc. Genetically modified microbes producing isoprenoids
JP4821886B2 (ja) 2009-06-04 2011-11-24 トヨタ自動車株式会社 組換え酵母、当該組換え酵母を用いた分岐アルコールの製造方法
EP2292741A1 (en) 2009-08-26 2011-03-09 OrganoBalance GmbH Genetically modified organisms for the production of lipids
EP2477478B1 (en) 2009-09-15 2016-11-30 Sapphire Energy, Inc. A system for transformation of the chloroplast genome of scenedesmus sp. and dunaliella sp.
BR112012011073B8 (pt) 2009-11-10 2023-01-10 Massachusetts Inst Technology Método para aumentar a produção de terpenoide em uma célula que produz um ou mais terpenoides
WO2011123567A1 (en) 2010-03-31 2011-10-06 Codexis, Inc. Production of geranyl diphosphate
JP2013526277A (ja) 2010-05-07 2013-06-24 グリーンライト バイオサイエンシーズ インコーポレーテッド 酵素リロケーションにより代謝経路のフラックスを制御する方法
EP2571991A1 (en) 2010-05-20 2013-03-27 Evolva SA Method of producing isoprenoid compounds in yeast
WO2012016172A2 (en) 2010-07-30 2012-02-02 Amyris, Inc. Genetically modified microbes producing increased levels of acetyl-coa derived compounds
WO2012016177A2 (en) 2010-07-30 2012-02-02 Amyris, Inc. Genetically modified microbes producing increased levels of acetyl-coa derived compounds
EP2444415A1 (en) 2010-10-20 2012-04-25 Genoplante-Valor 1-Deoxy-D-xylulose 5-phosphate synthase alleles responsible for enhanced terpene biosynthesis
EP2660327A4 (en) 2010-12-28 2016-04-13 Suntory Holdings Ltd METHOD OF USE OF A MONOTERPEN GLYCOSYL TRANSFERASE
PL2681313T3 (pl) 2011-02-28 2016-11-30 Komórka drożdżowa do wytwarzania terpenów i jej zastosowania
WO2012135591A2 (en) 2011-03-30 2012-10-04 Amyris, Inc. Microbial isoprenoid production using a heterologous dxp pathway
AU2012249320A1 (en) 2011-04-29 2013-11-14 Danisco Us Inc. Recombinant microorganisms for enhanced production of mevalonate, isoprene, and isoprenoids
CA2832979C (en) 2011-05-09 2020-04-14 Amyris, Inc. Production of acetyl-coenzyme a derived compounds
BR112014002745A8 (pt) 2011-08-05 2017-06-20 Danisco Us Inc produção de isoprenoides sob condições de ph neutro
US9382553B2 (en) 2011-08-16 2016-07-05 The Regents Of The University Of California Host cells and methods for producing 1-deoxyxylulose 5-phosphate (DXP) and/or a DXP derived compound
EP2764108A2 (en) 2011-10-07 2014-08-13 Danisco US Inc. Methods for increasing microbial production of isoprene, isoprenoids, and isoprenoid precursor molecules using glucose and acetate co-metabolism
EP3339426B1 (en) 2011-10-07 2022-08-24 Danisco US Inc. Utilization of phosphoketolase in the production of mevalonate, isoprenoid precursors, and isoprene
BR112014010750B1 (pt) 2011-11-09 2021-09-21 Amyris, Inc Produção de isoprenoides derivados de acetil-coenzima a
WO2013096863A1 (en) 2011-12-23 2013-06-27 The Regents Of The University Of California Constructs and methods for improved isoprene biosynthesis
JP2013158298A (ja) 2012-02-06 2013-08-19 Suntory Holdings Ltd モノテルペン配糖体化酵素の利用方法
US10072279B2 (en) 2012-05-11 2018-09-11 Teknologian Tutkimuskeskus Vtt Oy Method for producing terpenes
PT2800811T (pt) 2012-05-25 2017-08-17 Univ California Métodos e composições para modificação de adn alvo dirigida por arn e para modulação dirigida por arn de transcrição
US20130323820A1 (en) 2012-06-01 2013-12-05 Lanzatech New Zealand Limited Recombinant microorganisms and uses therefor
EP2875136B1 (en) 2012-07-18 2017-04-12 Isobionics B.V. Rhodobacter for preparing terpenoids
US9382554B2 (en) 2012-07-27 2016-07-05 Wisys Technology Foundation, Inc. Methods for isoprene and pinene production in cyanobacteria
CA2879178C (en) 2012-08-07 2020-11-24 Amyris, Inc. Methods for stabilizing production of acetyl-coenzyme a derived compounds
BR112015003329A2 (pt) 2012-08-17 2017-09-26 Evolva Sa produção aumentada de terpenos e terpenoides
WO2014052054A1 (en) 2012-09-25 2014-04-03 Albert Einstein College Of Medicine Of Yeshiva University Enzymatic platform for the synthesis of isoprenoid precursors and uses thereof
WO2014066892A1 (en) 2012-10-26 2014-05-01 Coelho Pedro S De novo metabolic pathways for isoprene biosynthesis
WO2014089436A1 (en) 2012-12-07 2014-06-12 Ginkgo Bioworks, Inc. Methods and systems for methylotrophic production of organic compounds
ES2901396T3 (es) 2013-03-14 2022-03-22 Caribou Biosciences Inc Composiciones y métodos de ácidos nucleicos dirigidos a ácido nucleico
MY171958A (en) 2013-03-15 2019-11-08 Amyris Inc Use of phosphoketolase and phosphotransacetylase for production of acetyl-coenzyme a derived compounds
EP2984105B1 (en) 2013-04-10 2019-05-22 Danisco US Inc. Phosphoketolases for improved production of acetyl coenzyme a-derived metabolites, isoprene, isoprenoid precursors, and isoprenoids
ES2915377T3 (es) 2013-08-02 2022-06-22 Enevolv Inc Procedimientos y células huésped para ingeniería genómica, de vías y biomolécular
CN105934517A (zh) 2013-08-07 2016-09-07 阿迈瑞斯公司 用于稳定生产乙酰辅酶a衍生化合物的方法
WO2015030681A1 (en) 2013-08-30 2015-03-05 National University Of Singapore In vitro synthetic multi-biocatalytic system for the synthesis of isoprenoids and isoprenoid precursors
US20150203880A1 (en) 2013-11-06 2015-07-23 Massachusetts Institute Of Technology Co-culture based modular engineering for the biosynthesis of isoprenoids, aromatics and aromatic-derived compounds
WO2015100180A1 (en) 2013-12-23 2015-07-02 Danisco Us Ing. Compositions and methods for control of industrial scale production of bio-products
US20150225743A1 (en) 2014-02-10 2015-08-13 E I Du Pont De Nemours And Company Biocatalyst for conversion of methane and methanol to isoprene
JP6744819B2 (ja) 2014-02-20 2020-08-19 ダニスコ・ユーエス・インク メバロン酸、イソプレン、イソプレノイド前駆体、イソプレノイド、およびアセチルＣｏＡ由来産物の生産向上のための組換え微生物
RU2014112066A (ru) 2014-03-28 2015-10-10 Адзиномото Ко., Инк. Способ получения мономерного изопрена
US10900060B2 (en) 2014-06-13 2021-01-26 Deinove Method of producing terpenes or terpenoids
WO2016008885A1 (en) 2014-07-14 2016-01-21 Photanol B.V. Biosynthesis of sesquiterpenes in cyanobacteria
WO2016008883A1 (en) 2014-07-14 2016-01-21 Photanol B.V. Biosynthesis of monoterpenes in cyanobacteria
US20170298363A1 (en) 2014-09-18 2017-10-19 Genomatica, Inc Non-natural microbial organisms with improved energetic efficiency
BR112017014223A2 (pt) 2014-12-29 2018-03-06 Futuragene Israel Ltd planta lenhosa geneticamente modificada, estrutura de ácido nucleico, sistema de estruturas de ácido nucleico, célula isolada, célula vegetal isolada, composição pesticida, método para reforço da resistência de uma planta lenhosa à infecção causada por pragas, método para reforço de, pelo menos, um teor de óleo de geraniol, geranial, neral, citronelol, citronelal e citral de uma planta lenhosa, método para produção de óleo, óleo, óleo de eucalipto, método para produzir, pelo menos, um monoterpeno, e planta geneticamente modificada
US10808015B2 (en) 2015-06-25 2020-10-20 Amyris, Inc. Maltose dependent degrons, maltose-responsive promoters, stabilization constructs, and their use in production of non-catabolic compounds
US9988624B2 (en) 2015-12-07 2018-06-05 Zymergen Inc. Microbial strain improvement by a HTP genomic engineering platform
US20190211364A1 (en) 2016-07-01 2019-07-11 International Flavors & Fragrances Inc. Compositions and methods for producing citrus terpenoids
WO2018111194A1 (en) 2016-12-16 2018-06-21 Temasek Life Sciences Laboratory Limited Targets for improving terpene production in rhodosporidium toruloides
KR20200095534A (ko) 2017-12-07 2020-08-10 지머젠 인코포레이티드 발효에 의한 (6e)-8-히드록시제라니올의 제조를 위해 조작된 생합성 경로
JP2021506316A (ja) 2017-12-21 2021-02-22 ザイマージェン インコーポレイテッド ネペタラクトールオキシドレダクターゼ、ネペタラクトールシンターゼ、及びネペタラクトンを産生する能力がある微生物

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101509005A (zh) * 2009-03-02 2009-08-19 山东大学 川西獐牙菜香叶醇-10羟化酶基因SmG10H及其应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
FIAMMETA ALAGNA等: "Olive phenolic compounds:metabolic and transcriptional profiling during fruit sevelopment" *
RENE HOFER等: "Geraniol hydroxylase and hydroxygeratiion oxidase activities of the CYP76 famiily of cytochrome P450 enzymes and potential for engineering the eary steps of the (seco)iridoid pathway" *
VASILEV NIKOLAY等: "Assessment of Cultivation Factors that Affect Biomass and Geraniol Production in Transgenic Tobacco Cell Suspension Cultures" *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113234607A (zh) * 2021-05-26 2021-08-10 华中农业大学 利用米曲霉合成香叶醇的方法
CN114621982A (zh) * 2022-03-16 2022-06-14 嘉兴欣贝莱生物科技有限公司 香叶基二磷酸酯的生物合成方法及其在制备大麻类化合物中的应用
CN114621982B (zh) * 2022-03-16 2023-11-07 嘉兴欣贝莱生物科技有限公司 香叶基二磷酸酯的生物合成方法及其在制备大麻类化合物中的应用
CN114717210A (zh) * 2022-04-08 2022-07-08 四川农业大学 一种杨树香叶基香叶醇还原酶及其编码基因与应用
CN114717210B (zh) * 2022-04-08 2023-07-04 四川农业大学 一种杨树香叶基香叶醇还原酶及其编码基因与应用

Also Published As

Publication number Publication date
CA3084263A1 (en) 2019-06-13
US20190338259A1 (en) 2019-11-07
JP2021505154A (ja) 2021-02-18
KR20200095534A (ko) 2020-08-10
WO2019113387A1 (en) 2019-06-13
EP3720944A1 (en) 2020-10-14
US10662415B2 (en) 2020-05-26

Similar Documents

Publication Publication Date Title
CN111699247A (zh) 通过发酵产生(6e)-8-羟基香叶醇的经改造的生物合成途径
US11028416B2 (en) Engineered biosynthetic pathways for production of tyramine by fermentation
US8703454B2 (en) Method for producing (+)-zizaene
KR20200057799A (ko) 스테비올 글리코시드의 재조합 생산
CN111868047A (zh) 荆芥内半缩醛氧化还原酶、荆芥内半缩醛合酶和能够产生荆芥内酯的微生物
US9714440B2 (en) Method for producing patchoulol and 7-epi-α-selinene
CN116368233A (zh) 通过发酵生产4-氨基苯乙胺的工程化生物合成途径
KR20210097723A (ko) 발효에 의한 1,5-디아미노펜탄의 생산을 위한 조작된 생합성 경로
CN112368387A (zh) 通过发酵产生组胺的经改造的生物合成途径
CN108473972B (zh) 补身醇合酶iii
CN114525215B (zh) 产萜类化合物的重组菌株及其构建方法和发酵产萜类化合物的方法及应用
CN114058560A (zh) 甘氨酸的生产方法
CN113330108A (zh) 通过发酵产生2-氧代己二酸的经改造的生物合成途径
CN113754785B (zh) 融合蛋白及其制备方法与在制备岩藻糖基化产物中的应用
US20230175023A1 (en) Engineered biosynthetic pathways for production of 3,4-dihydroxybenzoic acid by fermentation
WO2022173436A1 (en) Engineered biosynthetic pathways for production of ectoine by fermentation
WO2022245340A1 (en) Engineered biosynthetic pathways for production of deoxyhydrochorismic acid by fermentation

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
REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 40035041

Country of ref document: HK

WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20200922

WD01 Invention patent application deemed withdrawn after publication