CN1124347C - 2－氨基－4－甲膦酰丁酸抗性基因 - Google Patents

2－氨基－4－甲膦酰丁酸抗性基因 Download PDF

Info

Publication number
CN1124347C
CN1124347C CN98115017A CN98115017A CN1124347C CN 1124347 C CN1124347 C CN 1124347C CN 98115017 A CN98115017 A CN 98115017A CN 98115017 A CN98115017 A CN 98115017A CN 1124347 C CN1124347 C CN 1124347C
Authority
CN
China
Prior art keywords
ptc
ptt
resistance
gene
amino
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.)
Expired - Lifetime
Application number
CN98115017A
Other languages
English (en)
Other versions
CN1225393A (zh
Inventor
埃克哈德·斯特罗克
沃夫冈·霍勒本
沃特·阿诺德
里纳特·阿利亚
阿里德·普勒
格哈德·沃纳
鲁迪格·马奎特
苏三尼·格拉布利
迪特·布罗尔
克劳斯·巴彻
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.)
Hechester Jsc
Hoechst AG
Original Assignee
Hechester Jsc
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
Priority claimed from DE19863628747 external-priority patent/DE3628747A1/de
Priority claimed from DE19863642829 external-priority patent/DE3642829A1/de
Priority claimed from DE19873700313 external-priority patent/DE3700313A1/de
Application filed by Hechester Jsc filed Critical Hechester Jsc
Publication of CN1225393A publication Critical patent/CN1225393A/zh
Application granted granted Critical
Publication of CN1124347C publication Critical patent/CN1124347C/zh
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
    • C12P41/006—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures
    • C12P41/007—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures by reactions involving acyl derivatives of racemic amines
    • 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
    • C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
    • 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
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
    • C12N15/8209—Selection, visualisation of transformants, reporter constructs, e.g. antibiotic resistance markers
    • 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
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance
    • C12N15/8277—Phosphinotricin
    • 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.)
    • C12N9/1025—Acyltransferases (2.3)
    • C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Saccharide Compounds (AREA)
  • Steroid Compounds (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Transition And Organic Metals Composition Catalysts For Addition Polymerization (AREA)
  • Catalysts (AREA)
  • Macromolecular Compounds Obtained By Forming Nitrogen-Containing Linkages In General (AREA)
  • Compounds Of Alkaline-Earth Elements, Aluminum Or Rare-Earth Metals (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

通过在绿色产色链霉菌DSM4112中选择2-氨基-4-甲膦酰-丙氨酰-丙氨酸(PTT)抗性而产生PTT抗性的选择体。用BamHI酶切这些选择体的总DNA得到携有Phosphinothricin(PTC)抗性基因的DNA片段，克隆4.0Kb大小的片段并选择PTT抗性。此基因适用于生产PTC抗性的植物，以及作为抗性标记用于L型外消旋PTC的选择性N-乙酰化作用。

Description

2-氨基-4-甲膦酰丁酸抗性基因
(一)发明领域
Phosphinothricin(PTC，2-氨基-4-甲膦酰丁酸)是谷氨酰胺合成酶的抑制剂。PTC是抗生素2-氨基-4-甲膦酰-丙氨酰-丙氨酸的“结构单位”。此三肽(PTT)具有抗革兰氏阳性和革兰氏阴性细菌以及抗真菌灰葡萄孢(Botrytis cinerea)的活性(Bayer等人，Helv.Chim.Acta 55：224，1972)。PTT是由绿色产色链霉菌(Streptomyces viridochromogenes)Tu 494菌株(D SM40736，D SM4112)产生的。
(二)背景技术
德国专利2,717,440号公开了PTC可起全除莠剂(total herbicide)的作用。已公开的PCT申请WO86/02097号描述了植物对PTC的抗性是归因于谷氨酰胺合成酶的过量生成。这种类型的过量生成(如从基因扩增所导致的)易遭受不稳定的风险。因此，这样一种不稳定性将带来谷氨酰胺合成酶过量生成的下降，将致使重新出现PTC的竞争性抑制作用。与之相反，在权利要求中所限定的本发明涉及一种PCT抗性基因，该基因可用于生产PTC抗性植物.此外，该基因还可作为抗性标记使用.而且，该基因可适于L型外消旋PTC的选择性N-乙酰化作用
(三)发明内容
本发明的PTC抗性基因能由下述步骤得到：用BamH I酶切以PTT抗性选择出来的绿色产色链霉菌D SM4112菌株的总DNA，克隆大小为4.0Kb的片段，然后选择PTT抗性。限制性图谱(图1)详细描绘了该4.0Kb片段的特征。
对该4Kb片段部分进行克隆实验，以更精确地确定编码区的位置。此实验结果表明，抗性基因位于1.6Kb的SstII-SstI片段上(图1中位置0.55至2.15)。用BglII消化产生0.8Kb大小的片段，此片段掺入质粒中并转化变铅青链霉菌(S.lividans)后赋予PTT抗性。此抗性是由PTC的N-乙酰化产生的。
对此0.8Kb片段进行Maxam和Gilbert序列分析得到DNA序列I(见附页)。抗性基因的位置可从这一序列的开放读码(open readingframe)确定(从位置258起)。此基因读码终点位于图示的倒数第二个核苷酸(位置806)，即最后一个核苷酸(位置807)是终止密码子的第一个核苷酸。
用下加线条强调标明了DNA序列I中的Shine-Dalgarno序列，作为起始密码子的GTG也用线条标出。而且，顶部线条标出了确定的读码。
DNA序列II显示已确定了序列的基因中的限制性切点。没有标出切断此序列六次以上的酶。
抗生素PTT由细菌摄取并被降解为PTC。后者也抑制细菌中的谷氨酰胺合成酶，致使细菌因缺乏谷氨酰胺而死亡。因此，产生PTT的细菌应当具有保护自身不受PTT影响的机理，也就是说，防止再度摄取已生产的PTT，或者对已降解的产物PTC进行修饰。但令人吃惊的是，PTT生产菌绿色产色链霉菌DSM4112对其自身的抗生素敏感。但出人意料地证实，通过选择PTT抗性，有可能以10-5的极高频率发现抗PTT的选择体，而且，此选择体还能抑制相邻菌落的本底生长。
通过分离此DNA、用BamH I切断并连接到链霉菌载体中建立了这些选择体DNA的基因库。将连接混合物转化到市售的变铅青链霉菌TK23菌株中，每1μg连接混合物产生大约5000至10000个含有大约1至5Kb插入段的转化体。在这些转化体中有PTT抗性变铅青链霉菌株。有可能通过质粒分离并重新转化到变铅青链霉菌中，证明该抗性是由质粒编码的。负责此抗性的基因位于4Kb BamH I片段上(图1)。编码区位于0.8Kb BglII片段上。BamH I片段中不含下列酶的切点： Cla I、EcoR I、 EcoR V、 Hind III、 Hpa I、 Kpn I、 Pvu I、 Pvu II和Xho I。
将抗性基因的限制性图谱(还没有详细确定其特征)与吸水链霉菌(S.hygroscopicus)FERM BP-130/ATCC21705(欧洲专利申请，公开号0,173,327，图7)相比较表明，本发明的抗性基因不同于已知基因，其为寻找PTT生物合成基因过程中发现的。
一方面使绿色产色链霉菌D SM4112和变铅青链霉菌TK23的细胞提取物与PTC和乙酰辅酶A保温，另一方面使PTT抗性的绿色产色链霉菌选择体和携有质粒的变铅青链霉菌转化体与PTC和乙酰辅酶A保温，有可能表明后面的细胞将具有乙酰化活性。层析分析表明乙酰化发生在氨基上。
由于大肠杆菌中也已发现了PTT抗性，而且抗性机理也在革兰氏阴性细菌中起作用，因此，有可能基于运送现象排除抗性。这样，与植物启动子偶联并使用合适的载体后，本发明的抗性基因可转化到植物中，从而可产生PTC抗性植物。
PTC的N-乙酰化还可用于合成的D，L-PTC的外消旋体拆开，因为只有L型发生选择性乙酰化作用。
因此，本发明还涉及使用抗性基因选择性地使外消旋PTC的L型发生N-乙酰化。
因此，本发明的抗性基因编码的PTC乙酰转移酶可用于分离外消旋PTC，例如，使外消旋体经受此酶的乙酰化作用。可使由德国专利2,717,440的方法所得到的外消旋PTC成为光学对映体，因为此酶只选择性地攻击L型，而D型则保持不变。然后，根据二者性质的差别，通过已知方法可将这样得到的混合物分离开。
已公开了这样一种方法：使N-酰基-D，L-氨基酸与固定在适当载体上的酰基转移酶接触，选择性地释放L-氨基酸，它可以在酸化以后用不与水混溶的溶剂从与N-酰基-D-氨基酸的混合物中萃取出来(见英国专利1,369,462号)。已经公开了N-酰基-D，L-PTC的相应的分离方法，如见于德国专利公开2,939,269号或美国专利4,226,941号。
根据本发明保持不变的D-PTC可通过已知方法外消旋(见公开号为EP-A-0,137,371的欧洲专利申请，实例8)，然后回到流程中。
有可能但并不一定要分离出酶，这里及下文中所说的“酶”总是意指具有酶促活性的部分。如果酶被分离出来，它可以以游离形式或在载体上的固定化形式使用。合适载体的例子已在欧洲专利-A0,141,223号中描述了。但方便的作法是不分离酶，而是使用任何预期可表达本发明之酶的PTC抗性细胞。因此，有可能方便地使用绿色产色链霉菌DSM4112的PTT抗性选择体。而且，有可能有利地使用任何用本发明之基因转化的预期细胞，并使之能够表达PTC乙酰转移酶。就此而论，本发明的基因(也是指其活性部分)可以以质粒整合形式或利用其它常规的基因操作方法，例如通过转染引入到宿主细胞中。例如，掺入大肠杆菌的表达质粒中并用这样的质粒转化大肠杆菌是方便可行的，例如可通过欧洲专利-A0,163,249号和0,171,024号中介绍的已知方法进行。
对于外消旋体中的L-PTC的N-乙酰化，根据本发明，可以游离或固定化形式使用表达PTC乙酰转移酶的细胞，使用常规的固定化技术进行固定(例如德国专利公开3,237,341及其中引用的文献)。
本发明中L-PTC的酶促乙酰化以常规的酶促反应方式进行，方法中的条件由所用生物的性质来确定。从原则上讲，适用于此的方法与上述选择性脱酰作用的方法相同。
(四)具体实施方式
下述实例进一步阐明了本发明。除非特别指出者外，其中的份数和百分数数据均是指重量。
实例1 PTT抗性选择体
在基本培养基中(Hopwood等人，《链霉菌的基因操作：实验室手册》，The John Innes Foundation Norwieh，England，p.233，1985)培养绿色产色链霉菌D SM4112菌株，加入渐增浓度的PTT。在100μg/ml的浓度下，大约在每105个菌落中发现一个抗性菌落。
实例2 载体的制备
用BglII切割质粒pSVHI(欧洲专利0,070,522号，美国专利4,673,642号)，分离大小约7.1Kb的片段并与1.1Kb具有硫链丝菌素抗性的BclI片段连接(欧洲专利申请，公开号0,158,201)。得到大小约为8.15Kb的质粒pEB2(图2)。
实例3 抗性基因的分离
从实例1得到的选择体中分离总DNA，并用BamHI酶切。同样用BamHI打开质粒pEB2，使两种混合物结合并连接。将连接混合物转化到变铅青链霉菌TK23(可从The John Innes Foundation得到)中，每1微克连接混合物得到5000至10000个具有大约1～5Kb插入段的转化体。选择PTT抗性产生两个具抗性的变铅青链霉菌菌落。从抗性菌落中分离出质粒并用BamHI酶切。发现一个携有负责抗性的基因的4Kb BamHI片段。此质粒被称为pPRI(图3)。
将其再转化到变铅青链霉菌TK23中，证明PTT抗性是由质粒编码的，因为转化体能在含有100μg/ml PTT的基体培养上生长。
实例4 证明N-乙酰化使PTC失活
检查下列菌株以证实克隆片段的乙酰化活性：绿色产色链霉菌DSM40736，绿色产色链霉菌(PTT抗性突变株)、变铅青链霉菌TK23和变铅青链霉菌TK23(pPRI)。
这需将菌株接种到溶解培养基A(欧洲专利申请，公开号0,158,872第6页)中，并在轨道式震荡培养器中于30℃保温2天。收获细胞后，取1mg菌丝体在合适的缓冲液(如RS缓冲液：C.J.Thompson等人，J.Bacteriol 151：678-685，1982)中用超声波破碎。测定PTC降解的典型试验程序如下：
在250μl粗提物中，加入100μl PTC溶液(250μg/ml)和50μl乙酰辅酶A(4mg/ml)，将混合物于30℃下保温2小时。此时间过后，用HPLC测定仍然存在的PTC量。此实验基结果如下：
                             未反应的PTC菌株
                             加入的PTC变铅青链霉菌TK23                 100％绿色产色链霉菌(D SM40736)        72％绿色产色链霉菌选择体             7％变铅青链霉菌TK23(pPRI)           31％
在薄层层析上(没有茚三酮显色)与参考物质相比表明，已经发生了PTC的N-乙酰化。
DNA序列IIleTrpSerAspValLeuGlyAlaGlyProValLeuProGlyAspAspPhePheSerLeuGlyGlyThrSerIleAspLeuGluArgArgProGlyGlyArgSerGlyAlaAlaArgGlyArgLeuLeuLeuProArgArgHisLeuHisArgSerGlyAlaThrSerTrpGlyProValArgCysCysProGlyThrThrSerSerProSerAlaAlaProProAGATCTGGAGCGACGTCCTGGGGGCCGGTCCGGTGCTGCCCGGGGACGACTTCTTCTCCCTCGGCGGCACCTCCA 75TCTAGACCTCGCTGCAGGACCCCCGGCCAGGCCACGACGGGCCCCTGCTGAAGAAGAGGGAGCCGCCGTGGAGGTSerArgSerArgArgGlyProProArgAspProAlaAlaArgProArgSerArgArgGlyArgArgCysArgTrpAspProAlaValAspGlnProGlyThrArgHisGlnGlyProValValGluGluGlyGluAlaAlaGlyGlyAspIleGlnLeuSerThrArgProAlaProGlyThrSerGlyProSerSerLysLysGluArgProProValGluMetSerAlaLeuArgValValSerArglleArgLysGluLeuGlyValProLeuArgLeuAlaValIlePheGluThrLeuGlyValAlaGlyGlyLeuAlaHisProGlnGlyThrArgArgAlaThrProAlaArgArgAspLeuArgAspSerArgArgCysGlyTrpSerArgAlaSerAlaArgAsnSerAlaCysHisSerGlySerProOP SerSerArgTCTCGGCGTTGCGGGTGGTCTCGCGCATCCGCAAGGAACTCGGCGTGCCACTCCGGCTCGCCGTGATCTTCGAGA 150AGAGCCGCAACGCCCACCAGAGCGCGTAGGCGTTCCTTGAGCCGCACGGTGAGGCCGAGCGGCACTAGAAGCTCTArgProThrAlaProProArgAlaCysGlyCysProValArgArgAlaValGlyAlaArgArgSerArgArgSerArgArgGlnProHisAspArgAlaAspAlaLeuPheGluAlaHisTrpGluProGluGlyHisAspGluLeuArgGluAlaAsnArgThrThrGluArgMetArgLeuSerSerProThrGlySerArgSerAlaThrIleLysSerValProSerLeuGluAlaValAlaGluSerValLeuArgGluLeuLysGlyThrAM OC ArgGlyAlaArgHisProAlaValProGlySerGlyGlyArgIleArgThrProArgThrGluGlyAspValValLysArgCysProProProArgArgProTrpLysArgTrpProAsnProTyrSerAlaAsnOP ArgGlyArgSerLysGluValProAlaThrCGCCGTCCCTGGAAGCGGTGGCCGAATCCGTACTCCGCGAACTGAAGGGGACGTAGTAAAGAGGTGCCCGCCACC 225GCGGCAGGGACCTTCGCCACCGGCTTAGGCATGAGGCGCTTGACTTCCCCTGCATCATTTCTCCACGGGCGGTGGAlaThrGlyProLeuProProArgIleArgValGlyArgValSerProSerThrThrPheLeuHisGlyGlyGlyArgGlyGlnPheArgHisGlyPheGlyTyrGluAlaPheGlnLeuProArgLeuLeuSerThrGlyAlaValArgGlyAspArgSerAlaThrAlaSerAspThrSerArgSerSerPheProValTyrTyrLeuProAlaArgTrpGlyDNA序列I(续前页)LeuSerGlnAsnThrGluGlyArgProHisMetSerProGluArgArgProValGluIleArgProAlaThrAlaAlaPheAlaGluHisArgArgLysThrThrArgGluProArgThrThrProGlyArgAspProSerArgHisArgArgPheArgArgThrProLysGluAspHisThrOP AlaGlnAsnAspAlaArgSerArgSerValProProProCGCTTTCGCAGAACACCGA AGGAAGACCACAC GTGAGCCCAGAACGACGCCCGGTCGAGATCCGTCCCGCCACCG 300GCGAAAGCGTCTTGTGGCTTCCTTCTGGTGTGCACTCGGGTCTTGCTGCGGGCCAGCTCTAGGCAGGGCGGTGGCAlaLysAlaSerCysArgLeuPheValValArgSerGlyLeuValValGlyProArgSerGlyAspArgTrpArgLysArgLeuValGlyPheSerSerTrpValHisAlaTrpPheSerAlaArgAspLeuAspThrGlyGlyGlyGlySerGluCysPheValSerProLeuGlyCysThrLeuGlySerArgArgGlyThrSerIleArgGlyAlaValAlaAlaAspMetAlaAlaValCysAspIleValAsnHisTyrIleGluThrSerThrValAsnPheArgThrGluProArgArgHisGlyGlyGlyLeuArgHisArgGlnSerLeuHisArgAspGluHisGlyGlnLeuProTyrGlyAlaProProThrTrpArgArgSerAlaThrSerSerIleThrThrSerArgArgAlaArgSerThrSerValArgSerCCGCCGACATGGCGGCGGTCTGCGACATCGTCAATCACTACATCGAGACGAGCACGGTCAACTTCCGTACGGAGC 375GGCGGCTGTACCGCCGCCAGACGCTGTAGCAGTTAGTGATGTAGCTCTGCTCGTGCCAGTTGAAGGCATGCCTCGArgArgCysProProProArgArgCysArgOP AspSerCysArgSerSerCysProOP SerGlyTyrProAlaGlyValHisArgArgAspAlaValAspAspIleValValAspLeuArgAlaArgAspValGluThrArgLeuArgAlaSerMetAlaAlaThrGlnSerMetThrLeuOP AM MetSerValLeuValThrLeuLysArgValSerGlyGlnThrProGlnGluTrpIleAspAspLeuGluArgLeuGlnAspArgTyrProTrpLeuValAlaGluValGluAlaAspSerAlaGlyValAspArgArgProGlyAlaProProGlyProLeuProLeuAlaArgArgArgGlyGlyArgArgLeuArgArgSerGlySerThrThrTrpSerAlaSerArgThrAlaThrProGlySerSerProArgTrpCGCAGACTCCGCAGGAGTGGATCGACGACCTGGAGCGCCTCCAGGACCGCTACCCCTGGCTCGTCGCCGAGGTGG 450GCGTCTGAGGCGTCCTCACCTAGCTGCTGGACCTCGCGGAGGTCCTGGCGATGGGGACCGAGCAGCGGCTCCACCAlaSerGluAlaProThrSerArgArgGlyProAlaGlyGlyProGlySerGlyArgAlaArgArgArgProProLeuSerArgLeuLeuProAspValValGlnLeuAlaGluLeuValAlaValGlyProGluAspGlyLeuHisLeuCysValGlyCysSerHisIleSerSerArgSerArgArgTrpSerArgAM GlyGlnSerThrAlaSerThrSerDNA序列I(续前页)GlyValValAlaGlyIleAlaTyrAlaGlyProTrpLysAlaArgAsnAlaTyrAspTrpThrValGluSerThrGlyArgArgArgArgHisArgLeuArgArgProLeuGluGlyProGlnArgLeuArgLeuAspArgArgValAspArgAlaSerSerProAlaSerProThrProAlaProGlyArgProAlaThrProThrThrGlyProSerSerArgAGGGCGTCGTCGCCGGCATCGCCTACGCCGGCCCCTGGAAGGCCCGCAACGCCTACSACTGGACCGTCGAGTCGA 525TCCCGCAGCAGCGGCCGTAGCGGATGCGGCCGGGGACCTTCCGGGCGTTGCGGATGCTGACCTGGCAGCTCAGCTProArgArgArgArgCysArgArgArgArgGlyArgSerProGlyCysArgArgArgSerSerArgArgThrSerAlaAspAspGlyAlaAspGlyValGlyAlaGlyProLeuGlyAlaValGlyValValProGlyAspLeuArgArgProThrThrAlaFroMetAlaAM AlaProGlyGlnPheAlaArgLeuAlaAM SerGlnValThrSerAspValValTyrValSerHisArgHisGlnArgLeuGlyLeuGlySerThrLeuTyrThrHisLeuLeuLysSerMetGluGlyValArgLeuProProAlaProAlaAlaArgThrGlyLeuHisProLeuHisProProAlaGluValHisGlyArgCysThrSerProThrGlyThrSerGlySerAspTrpAlaProProSerThrProThrCysOP SerProTrpCGGTGTACGTCTCCCACCGGCACCAGCGGCTCGGACTGGGCTCCACCCTCTACACCCACCTGCTGAAGTCCATGG 600GCCACATGCAGAGGGTGGCCGTGGTCGCCGAGCCTGACCCGAGGTGGGAGATGTGGGTGGACGACTTCAGGTACCProThrArgArgGlyGlyAlaGlyAlaAlaArgValProSerTrpGlyArgCysGlyGlyAlaSerThrTrpProHisValAspGlyValProValLeuProGluSerGlnAlaGlyGlyGluValGlyValGlnGlnLeuGlyHisLeuThrTyrThrGluTrpArgCysTrpArgSerProSerProGluValArgAM ValTrpArgSerPheAspMetSerAlaGlnGlyPheLysSerValValAlaValIleGlyLeuProAsnAspProSerValArgLeuHisGluAlaLeuGlyProGlyLeuGlnGluArgGlyArgArgHisArgThrAlaGlnArgProGluArgAlaProAlaArgGlyAlaArgProArgAlaSerArgAlaTrpSerProSerSerAspCysProThrThrArgAlaCysAlaCysThrArgArgAGGCCCAGGGCTTCAAGAGCGTGGTCGCCGTCATCGGACTGCCCAACGACCCGAGCGTGCGCCTGCACGAGGCGC 675TCCGGGTCCCGAAGTTCTCGCACCAGCGGCAGTAGCCTGACGGGTTGCTGGGCTCGCACGCGGACGTGCTCCGCGProGlyProSerOP SerArgProArgArgOP ArgValAlaTrpArgGlySerArgAlaGlyAlaArgProAlaGlyLeuAlaGluLeuAlaHisAspGlyAspAspSerGlnGlyValValArgAlaHisAlaGlnValLeuArgGluAlaTrpProLysLeuLeuThrThrAlaThrMetProSerGlyLeuSerGlyLeuThrArgArgCysSerAlaSerDNA序列I(续前页)GlyTyrThrAlaArgGlyThrLeuArgAlaAlaGlyTyrLysHisGlyGlyTrpHisAspValGlyPheTrpGlnArgIleHisArgAlaArgAspAlaAlaGlySerArgLeuGlnAlaArgGlyLeuAlaArgArgGlyValLeuAlaSerAspThrProArgAlaGlyArgCysGlyGlnProAlaThrSerThrGlyAlaGlyThrThrTrpGlySerGlyTCGGATACACCGCGCGCGGGACGCTGCGGGCAGCCGGCTACAAGCACGGGGGCTGGCACGACGTGGGGTTCTGGC 750AGCCTATGTGGCGCGCGCCCTGCGACGCCCGTCGGCCGATGTTCGTGCCCCCGACCGTGCTGCACCCCAAGACCGArgIleCysArgAlaArgSerAlaAlaProLeuArgSerCysAlaArgProSerAlaArgArgProThrArgAlaSerValGlyArgAlaProArgGlnProCysGlyAlaValLeuValProAlaProValValHisProGluProLeuProTyrValAlaArgProValSerArgAlaAlaProAM LeuCysProProGlnCysSerThrProAsnGlnCysArgAspPheGluLeuProAlaProProArgProValArgProValThrGlnIleAlaArgLeuArgAlaAlaGlyProAlaProProArgProAlaArgHisThrAspSerAlaThrSerSerCysArgProArgProAlaProSerGlyProSerHisArgSerAGCGCGACTTCGAGCTGCCGGCCCCGCCCCGCCCCGTCCGGCCCGTCACACAGATCT 807TCGCGCTGAAGCTCGACGGCCGGGGCGGGGCGGGGCAGGCCGGGCAGTGTGTCTAGAAlaArgSerArgAlaAlaProGlyAlaGlyGlyArgGlyAlaArgOP ValSerArgAlaValGluLeuGlnArgGlyArgGlyAlaGlyAspProGlyAspCysLeuAspArgSerLysSerSerGlyAlaGlyGlyArgGlyThrArgGlyThrValCysIleDNA序列II1 AGATCTGGAGCGACGTCCTGGGGGCCGGTCCGGTGCTGCCCGGGGACGACTTCTTCTCCC
TCTAGACCTCGCTGCAGGACCCCCGGCCAGGCCACGACGGGCCCCTGCTGAAGAAGAGGG
                                                ^
1 BGLII XHOII，2 DPNI SAU3A，5 GSUI，12 AATII ACYI，13 MAEII
，17 APYI ECORII，26 RSRII，27 AVAII，35 BBVI，39 AVAI NCII
SMAI，40 NCII，52 MBOII，59 MNLI，61 TCGGCGGCACCTCCATCTCGGCGTTGCGGGTGGTCTCGCGCATCCGCAAGGAACTCGGCG
AGCCGCCGTGGAGGTAGAGCCGCAACGCCCACCAGAGCGCGTAGGCGTTCCTTGAGCCGC
                                    ^^^^^
66 HGICI，70 MNLI，97 FNUDII，100 SFANI，101 FOKI，121 TGCCACTCCGGCTCGCCGTGATCTTCGAGACGCCGTCCCTGGAAGCGGTGGCCGAATCCG
ACGGTGAGGCCGAGCGGCACTAGAAGCTCTGCGGCAGGGACCTTCGCCACCGGCTTAGGC
                                                     ^^
122 BGLI，140 DPNI SAU3A，142 MBOII，149 ACYI HGAI TTH111I，
158 APYI ECORII，169 CFRI GDIII，174 HINFI，180 RSAI，181 TACTCCGCGAACTGAAGGGGACGTAGTAAAGAGGTGCCCGCCACCCGCTTTCGCAGAACA
ATGAGGCGCTTGACTTCCCCTGCATCATTTCTCCACGGGCGGTGGGCGAAAGCGTCTTGT
                             ^^^^^
186 FNUDII，201 MAEII，211 MNLI，213 HGICI，214 SDUI，241 CCGAAGGAAGACCACACGTGAGCCCAGAACGACGCCCGGTCGAGATCCGTCCCGCCACCG
GGCTTCCTTCTGGTGTGCACTCGGGTCTTGCTGCGGGCCAGCTCTAGGCAGGGCGGTGGC
                                    ^^^
247 MBOII，254 AFLIII，255 PMACI，256 MAEII，260 HGIJII SDUI
，271 ACYI HGAI，275 NCII，283 XHOII，284 BINI DPNI SAU3A，301 CCGCCGACATGGCGGCGGTCTGCGACATCGTCAATCACTACATCGAGACGAGCACGGTCA
GGCGGCTGTACCGCCGCCAGACGCTGTAGCAGTTAGTGATGTAGCTCTGCTCGTGCCAGT
                                                    ^^^^^
303 BGLI，308 NLAIII，324 TTH111I，350 HGIAI SDUI，357 HINCI
I，361 ACTTCCGTACGGAGCCGCAGACTCCGCAGGAGTGGATCGACGACCTGGAGCGCCTCCAGG
TGAAGGCATGCCTCGGCGTCTGAGGCGTCCTCACCTAGCTGCTGGACCTCGCGGAGGTCC
                                                 ^^^^^
367 RSAI，380 HINFI，394 BINI，395 DPNI SAU3A，404 APYI ECOR
II，405 GSUI，409 HAEII，413 MNLI，414 GSUI，416 APYI ECORII
，419 AVAII，DNA序列II(续前页)421 ACCGCTACCCCTGGCTCGTCGCCGAGGTGGAGGGCGTCGTCGCCGGCATCGCCTACGCCG
TGGCGATGGGGACCGAGCAGCGGCTCCACCTCCCGCAGCAGCGGCCGTAGCGGATGCGGC
                                                  ^^
430 APYI ECORII，444 MNLI，450 MNLI，453 ACYI，454 HGAI，462
NAEI，4GG·SFANI，477 NAEI，481 GCCCCTGGAAGGCCCGCAACGCCTACGACTGGACCGTCGAGTCGACGGTGTACGTCTCCC
CGGGGACCTTCCGGGCGTTGCGGATGCTGACCTGGCAGCTCAGCTGCCACATGCAGAGGG
484 APYI ECORII，S11 AVAII，519 HINFI，521 ACCI HINCII SALI，
 530 RSAI，532 MAEII，541 ACCGGCACCAGCGGCTCGGACTGGGCTCCACCCTCTACACCCACCTGCTGAAGTCCt
TGGCCGTGGTCGCCGAGCCTGACCCGAGGTGGGAGATGTGGGTGGACGACTTCAGGTt
                                                 ^^
544 HGICI，549 NSPBII，563 HGIJII SDUI，572 MNLI，578 TAQII，
 563 BSPMI，595 NCOI STYI，596 NLAIII，600 MNLI，601 AGGCCCAGGGCTTCAAGAGCGTGGTCGCCGTCATCGGACTGCCCAACGACCCGAGCGTGC
TCCGGGTCCCGAAGTTCTCGCACCAGCGGCAGTAGCCTGACGGGTTGCTGGGCTCGCACG
                                                ^^
605 APYI ECORII，650 AVAI，661 GCCTGCACGAGGCGCTCGGATACACCGCGCGCGGGACGCTGCGGGCAGCCGGCTACAAGC
CGGACGTGCTCCGCGAGCCTATGTGGCGCGCGCCCTGCGACGCCCGTCGGCCGATGTTCG
                                              ^^^^^
669 MNLI，671 HAEII，686 FNUDII，687 BSSHII，688 FNUDII，690
 FNUDII，695 HGAI，698 BBVI，705 BBVI，708 NAEI，716 TTH111I
I，721 ACGGGGGCTGGCACGACGTGGGGTTCTGGCAGCGCGACTTCGAGCTGCCGGCCCCGCCCC
TGCCCCCGACCGTGCTGCACCCCAAGACCGTCGCGCTGAAGCTCGACGGCCGGGGCGGGG
                                         ^
732 DRAIII，736 MAEII，749 BBVI，753 FNUDII，763 ALUI，764 B
BVI，767 NAEI，781 GCCCCGTCCGGCCCGTCACACAGATCT
CGGGGCAGGCCGGGCAGTGTGTCTAGA
                    ^^^
795 MAEIII，802 BGLII XHOII，803 DPNI SAU3A，

Claims (1)

1.2-氨基-4-甲膦酰丁酸抗性基因，该基因编码以下所示的氨基酸序列：
                          MetSerProGluArgArgProValGluIleArgProAlaThrAlaAlaAspMetAlaAlaValCysAspIleValAsnHisTyrIleGluThrSerThrValAsnPheArgThrGluProGlnThrProGlnGluTrpIleAspAspLeuGluArgLeuGlnAspArgTyrProTrpLeuValAlaGluValGluGlyValValAlaGlyIleAlaTyrAlaGlyProTrpLysAlaArgAsnAlaTyrAspTrpThrValGluSerThrValTyrValSerHisArgHisGlnArgLeuGlyLeuGlySerThrLeuTyrThrHisLeuLeuLysSerMelGluAlaGlnGlyPheLysSerValValAlaValIleGlyLeuProAsnAspProSerValArgLeuHisGluAlaLeuGlyTyrThrAlaArgGlyThrLeuArgAlaAlaGlyTyrLysHisGlyGlyTrpHisAspValGlyPheTrpGlnArgAspPheGluLeuProAlaProProArgProValArgProValThrGlnI1e.
CN98115017A 1986-08-23 1998-06-19 2－氨基－4－甲膦酰丁酸抗性基因 Expired - Lifetime CN1124347C (zh)

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
DEP3628747.4 1986-08-23
DE19863628747 DE3628747A1 (de) 1986-08-23 1986-08-23 Resistenzgen gegen phosphinothricin und seine verwendung
DE3637307 1986-11-03
DEP3637307.9 1986-11-03
DE19863642829 DE3642829A1 (de) 1986-08-23 1986-12-16 Resistenzgen gegen phosphinothricin
DEP3642829.9 1986-12-16
DEP3700313.5 1987-01-08
DE19873700313 DE3700313A1 (de) 1986-08-23 1987-01-08 Verwendung eines resistenzgens gegen phosphinothricin

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN87105764A Division CN1040772C (zh) 1986-08-23 1987-08-22 获得2－氨基－4－甲膦酰丁酸基因的方法

Publications (2)

Publication Number Publication Date
CN1225393A CN1225393A (zh) 1999-08-11
CN1124347C true CN1124347C (zh) 2003-10-15

Family

ID=27433686

Family Applications (2)

Application Number Title Priority Date Filing Date
CN87105764A Expired - Lifetime CN1040772C (zh) 1986-08-23 1987-08-22 获得2－氨基－4－甲膦酰丁酸基因的方法
CN98115017A Expired - Lifetime CN1124347C (zh) 1986-08-23 1998-06-19 2－氨基－4－甲膦酰丁酸抗性基因

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN87105764A Expired - Lifetime CN1040772C (zh) 1986-08-23 1987-08-22 获得2－氨基－4－甲膦酰丁酸基因的方法

Country Status (13)

Country Link
EP (1) EP0257542B1 (zh)
JP (4) JPH0797994B2 (zh)
CN (2) CN1040772C (zh)
AT (1) ATE75776T1 (zh)
AU (1) AU604743B2 (zh)
CA (1) CA1337597C (zh)
DK (1) DK175254B1 (zh)
ES (1) ES2038631T3 (zh)
FI (1) FI100251B (zh)
GR (1) GR3005200T3 (zh)
HU (1) HU217208B (zh)
IL (1) IL83604A (zh)
NZ (1) NZ221526A (zh)

Families Citing this family (31)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3765449D1 (de) * 1986-03-11 1990-11-15 Plant Genetic Systems Nv Durch gentechnologie erhaltene und gegen glutaminsynthetase-inhibitoren resistente pflanzenzellen.
CN87100603A (zh) * 1987-01-21 1988-08-10 昂科公司 抗黑素瘤疫苗
DE3716309A1 (de) * 1987-05-15 1988-11-24 Hoechst Ag Resistenzgen gegen phosphinothricin
DE3732972A1 (de) * 1987-07-02 1989-01-12 Hoechst Ag Resistenzgen gegen phosphinothricin und seine verwendung
DE4126414A1 (de) * 1991-08-09 1993-02-11 Hoechst Ag Deacetylasegene zur erzeugung von phosphinothricin oder phosphinothricyl-alanyl-alanin, verfahren zu ihrer isolierung und ihre verwendung
DE4222407C1 (de) * 1992-07-08 1993-10-07 Max Planck Gesellschaft Modulartiges Promotor-Konstrukt
US6586367B2 (en) 1996-09-05 2003-07-01 Syngenta Crop Protection, Inc. Process for the control of weeds
AR008158A1 (es) * 1996-09-05 1999-12-09 Syngenta Participations Ag Proceso para el control de malas hierbas en cultivos de plantas utiles que son resistentes a un fosfo-herbicida y una composicion herbicida para dicho uso.
DE19836684A1 (de) 1998-08-13 2000-02-17 Hoechst Schering Agrevo Gmbh Herbizide Mittel für tolerante oder resistente Reiskulturen
DE19836700A1 (de) 1998-08-13 2000-02-17 Hoechst Schering Agrevo Gmbh Herbizide Mittel für tolerante oder resistente Getreidekulturen
DE19836659A1 (de) 1998-08-13 2000-02-17 Hoechst Schering Agrevo Gmbh Herbizide Mittel für tolerante oder resistente Baumwollkulturen
DE19836660A1 (de) 1998-08-13 2000-02-17 Hoechst Schering Agrevo Gmbh Herbizide Mittel für tolerante oder resistente Sojakulturen
PL218413B1 (pl) 1998-08-13 2014-12-31 Bayer Cropscience Ag Zastosowanie kompozycji zawierającej glifosat oraz chlopyralid do zwalczania szkodliwych roślin w uprawach kukurydzy oraz sposób zwalczania szkodliwych roślin w tolerujących uprawach kukurydzy
CA2475485C (en) 2002-02-26 2012-08-28 Syngenta Limited A method of selectively producing male or female sterile plants
CA2586241C (en) 2004-11-17 2013-07-30 Hokko Chemical Industry Co., Ltd. Herbicide-resistance gene and utilization thereof
KR100743611B1 (ko) * 2006-03-28 2007-08-01 최광술 산업용 로봇의 케이블 조절장치
AR074941A1 (es) 2009-01-07 2011-02-23 Bayer Cropscience Sa Plantas transplastomicas exentas de marcador de seleccion
CN102869769A (zh) 2009-12-23 2013-01-09 拜尔知识产权有限公司 耐受hppd抑制剂型除草剂的植物
EP2516630B1 (en) 2009-12-23 2017-11-15 Bayer Intellectual Property GmbH Plants tolerant to hppd inhibitor herbicides
EP2516631B1 (en) 2009-12-23 2018-02-14 Bayer Intellectual Property GmbH Plants tolerant to hppd inhibitor herbicides
BR112012015697A2 (pt) 2009-12-23 2015-08-25 Bayer Intelectual Property Gmbh Plantas tolerantes a herbicidas inibidores de hppd.
WO2011076889A1 (en) 2009-12-23 2011-06-30 Bayer Cropscience Ag Plants tolerant to hppd inhibitor herbicides
WO2011144684A1 (de) 2010-05-21 2011-11-24 Bayer Cropscience Ag Herbizide mittel für tolerante oder resistente reiskulturen
WO2011144683A1 (de) 2010-05-21 2011-11-24 Bayer Cropscience Ag Herbizide mittel für tolerante oder resistente rapskulturen
BR112012029616A2 (pt) 2010-05-21 2015-10-20 Bayer Ip Gmbh agentes herbicidas para as culturas de cereais tolerantes ou resistentes
BR112012029621A2 (pt) 2010-05-21 2015-09-22 Bayer Ip Gmbh agentes herbicidas para as culturas de milho tolerantes ou resistentes
WO2012004013A2 (en) 2010-07-08 2012-01-12 Bayer Bioscience N.V. Glucosinolate transporter protein and uses thereof
US20140090102A1 (en) 2011-01-24 2014-03-27 Stéphane Pien Use of the rd29 promoter or fragments thereof for stress-inducible expression of transgenes in cotton
EP2524602A1 (de) 2011-05-20 2012-11-21 Bayer CropScience AG Herbizide Mittel für tolerante oder resistente Sojakulturen
UA117816C2 (uk) 2012-11-06 2018-10-10 Байєр Кропсайєнс Акцієнгезелльшафт Гербіцидна комбінація для толерантних соєвих культур
DE202014101590U1 (de) 2014-04-03 2014-04-29 Igus Gmbh Führungssystem für Versorgungsleitungen und Roboter mit Führungssystem

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3765449D1 (de) * 1986-03-11 1990-11-15 Plant Genetic Systems Nv Durch gentechnologie erhaltene und gegen glutaminsynthetase-inhibitoren resistente pflanzenzellen.

Also Published As

Publication number Publication date
HU217208B (hu) 1999-12-28
DK437887A (da) 1988-02-24
JPS6371183A (ja) 1988-03-31
CN1225393A (zh) 1999-08-11
FI873610A0 (fi) 1987-08-20
EP0257542A2 (de) 1988-03-02
JPH03103193A (ja) 1991-04-30
IL83604A (en) 2004-02-19
CN1040772C (zh) 1998-11-18
CA1337597C (en) 1995-11-21
JP2749424B2 (ja) 1998-05-13
JPH0797994B2 (ja) 1995-10-25
FI100251B (fi) 1997-10-31
AU604743B2 (en) 1991-01-03
NZ221526A (en) 1989-03-29
AU7731887A (en) 1988-05-19
IL83604A0 (en) 1988-01-31
HUT44621A (en) 1988-03-28
FI873610A (fi) 1988-02-24
JPH07147985A (ja) 1995-06-13
EP0257542B1 (de) 1992-05-06
DK175254B1 (da) 2004-07-26
CN87105764A (zh) 1988-11-30
JP2815847B2 (ja) 1998-10-27
GR3005200T3 (zh) 1993-05-24
JPH09107981A (ja) 1997-04-28
ATE75776T1 (de) 1992-05-15
ES2038631T3 (es) 1993-08-01
EP0257542A3 (en) 1990-03-07
DK437887D0 (da) 1987-08-21

Similar Documents

Publication Publication Date Title
CN1124347C (zh) 2－氨基－4－甲膦酰丁酸抗性基因
CN106434651B (zh) 根癌农杆菌和CRISPR-Cas9介导的基因定点插入失活方法及其应用
US5276268A (en) Phosphinothricin-resistance gene, and its use
JP2674998B2 (ja) 抗生物質産生微生物の改良
US20090203139A1 (en) Production of four carbon alcohols using improved strain
KR102345899B1 (ko) 박테리아 헤모글로빈 라이브러리를 생성하는 방법 및 이의 용도
CN1225125A (zh) 用于将木糖发酵为乙醇的稳定的重组酵母
WO2019086708A1 (en) A genetically modified bacillus subtilis strain, optimized vectors, and uses thereof
WO2009117843A1 (zh) 敲除酰胺酶基因的产腈水合酶工程菌及其构建方法和应用
CN110157654B (zh) 一种纳豆芽孢杆菌重组菌及其构建方法与应用
DE3855633T2 (de) Rekombinante DNS-Expressionsvektoren und DNS-Zusammensetzungen, die für Deacetoxycephalosporin C-Synthetase und Deacetylcephalosporin C-Synthetase kodieren
CN112126615B (zh) 一种产丁酸的枯草芽孢杆菌及其构建方法和应用
RU2224022C2 (ru) Гены синтазы раффинозы и их использование
CN1055314C (zh) 从葡聚糖明串珠菌克隆和过量表达葡糖－6－磷酸脱氢酶
US20120015404A1 (en) Gene cluster for thuringiensin synthesis
EP0089233A2 (en) Plasmid pSAN 81 and a process for its extraction from a cell culture
CN109929853B (zh) 嗜热菌来源的热激蛋白基因的应用
CN108251447B (zh) 一种能高效表达脂肪酶的质粒、其构建方法及其应用
Poehlein et al. Complete Genome Sequence of Sporomusa termitida DSM 4440T
EP0757101B1 (en) Circular plasmids from Rhodococcus
CN113817761B (zh) 一种无三羧酸循环的大肠杆菌底盘菌及其构建方法与应用
CN115058440B (zh) 催化合成天然蔗糖酯的工程菌及其构建方法和应用
US20240110167A1 (en) Enzymes with ruvc domains
EP0284185A1 (en) A method for production of L-phenylalanine by recombinant E. coli
JP2009514506A (ja) Ｅ．コリ株ｄｓｍ６６０１のプラスミド不含のクローン

Legal Events

Date Code Title Description
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C06 Publication
PB01 Publication
C14 Grant of patent or utility model
GR01 Patent grant
C17 Cessation of patent right
CX01 Expiry of patent term