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1. 发明授权

US07608437B2 Method of constructing amino acid producing bacterial strains, and method of preparing amino acids by fermentation with the constructed amino acid producing bacterial strains 有权
标题翻译：构建产氨酸的细菌菌株的方法，以及用构建的产生氨基酸的菌株通过发酵制备氨基酸的方法
公开(公告)号：US07608437B2
公开(公告)日：2009-10-27
申请号：US11218433
申请日：2005-09-06
申请人： Yoko Asakura , Jun Nakamura , Sohei Kanno , Mikiko Suga , Eiichiro Kimura , Hisao Ito , Kazuhiko Matsui , Tsuyoshi Ohsumi , Tsuyoshi Nakamatsu , Osamu Kurahashi
发明人： Yoko Asakura , Jun Nakamura , Sohei Kanno , Mikiko Suga , Eiichiro Kimura , Hisao Ito , Kazuhiko Matsui , Tsuyoshi Ohsumi , Tsuyoshi Nakamatsu , Osamu Kurahashi
IPC分类号： C12P13/14 , C12Q1/48 , C12Q1/32 , C12Q1/68 , C12N9/02 , C12N9/10 , C12N1/21 , C12P21/00 , C12N15/00 , C07K14/00 , C07H21/00
CPC分类号： C12P13/14 , C12N15/67
摘要： A method of producing coryneform bacteria having an improved amino acid or nucleic acid-productivity comprises the steps of introducing a mutation in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium so that it is close to a consensus sequence or introducing a change in the promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on the chromosome of a coryneform bacterium by gene recombination so that it is close to a consensus sequence, obtaining mutants of the coryneform amino acid- or nucleic acid-producing microorganism, culturing the mutants and select a mutant capable of producing the intended amino acid or nucleic acid in a large amount. This method allows one of skill in the art to construct a mutant capable of enriching or controlling the expression of an intended gene without using a plasmid and to promote production of amino acids in a high yield, by the recombination or mutation.
摘要翻译：具有改进的氨基酸或核酸生产力的棒状杆菌细菌的方法包括以下步骤：将氨基酸或核酸 - 生物合成基因的启动子序列中的突变引入棒状细菌的染色体上，使其接近共有序列或通过基因重组在棒状细菌的染色体上引入氨基酸或核酸 - 生物合成基因的启动子序列的变化，使得其接近共有序列，获得棒状杆菌型氨基酸的突变体或产生核酸的微生物，培养突变体并选择能够大量产生所需氨基酸或核酸的突变体。该方法允许本领域技术人员通过重组或突变来构建能够富集或控制预期基因的表达而不使用质粒并以高产率促进氨基酸产生的突变体。

2. 发明申请

US20060003424A1 Method of constructing amino acid producing bacterial strains, and method of preparing amino acids by fermentation with the constructed amino acid producing bacterial strains 有权
标题翻译：构建产氨酸的细菌菌株的方法，以及用构建的产生氨基酸的菌株通过发酵制备氨基酸的方法
公开(公告)号：US20060003424A1
公开(公告)日：2006-01-05
申请号：US11218433
申请日：2005-09-06
申请人： Yoko Asakura , Jun Nakamura , Sohei Kanno , Mikiko Suga , Eiichiro Kimura , Hisao Ito , Kazuhiko Matsui , Tsuyoshi Ohsumi , Tsuyoshi Nakamatsu , Osamu Kurahashi
发明人： Yoko Asakura , Jun Nakamura , Sohei Kanno , Mikiko Suga , Eiichiro Kimura , Hisao Ito , Kazuhiko Matsui , Tsuyoshi Ohsumi , Tsuyoshi Nakamatsu , Osamu Kurahashi
IPC分类号： C12P13/22 , C12P13/14 , C12P13/08 , C12N1/21
CPC分类号： C12P13/14 , C12N15/67
摘要： A method of producing coryneform bacteria having an improved amino acid- or nucleic acid-productivity comprises the steps of introducing a mutation in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium to make it close to a consensus sequence or introducing a change in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium by gene recombination to make it close to a consensus sequence, to obtain mutants of the coryneform amino acid d- or nucleic acid-producing microorganism, culturing the mutants and select a mutant capable of producing the intended amino acid or nucleic acid in a large amount. This method can construct a mutant capable of suitably enriching or controling the expression of an intended gene without using a plasmid and also capable of producing amino acids in a high yield, by the recombination or mutation.
摘要翻译：具有改善的氨基酸或核酸生产力的棒状细菌的方法包括以下步骤：在棒状细菌的染色体上的氨基酸或核酸 - 生物合成基因的启动子序列中引入突变，使其接近共有序列或通过基因重组在棒状细菌的染色体上引入氨基酸或核酸 - 生物合成基因的启动子序列的变化，使其接近共有序列，获得棒状杆菌氨基酸d- 或产生核酸的微生物，培养突变体并选择能够大量产生所需氨基酸或核酸的突变体。该方法可以构建能够适当地富集或控制预期基因的表达而不使用质粒并且还能够通过重组或突变以高产率产生氨基酸的突变体。

3. 发明授权

US06962805B2 Method of constructing amino acid producing bacterial strains, and method of preparing amino acids by fermentation with the constructed amino acid producing bacterial strains 有权
标题翻译：构建产氨酸的细菌菌株的方法，以及用构建的产生氨基酸的菌株通过发酵制备氨基酸的方法
公开(公告)号：US06962805B2
公开(公告)日：2005-11-08
申请号：US09577005
申请日：2000-05-25
申请人： Yoko Asakura , Jun Nakamura , Sohei Kanno , Mikiko Suga , Eiichiro Kimura , Hisao Ito , Kazuhiko Matsui , Tsuyoshi Ohsumi , Tsuyoshi Nakamatsu , Osamu Kurahashi
发明人： Yoko Asakura , Jun Nakamura , Sohei Kanno , Mikiko Suga , Eiichiro Kimura , Hisao Ito , Kazuhiko Matsui , Tsuyoshi Ohsumi , Tsuyoshi Nakamatsu , Osamu Kurahashi
IPC分类号： C12N15/37 , C12N1/21 , C12N15/67 , C12P13/04 , C12P13/14 , C07H21/04 , C12N1/20 , C12N15/74 , C12Q1/32
CPC分类号： C12P13/14 , C12N15/67
摘要： A method of producing coryneform bacteria having improved amino acid or nucleic acid productivity comprising the steps of introducing a mutation in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium to make it close to a consensus sequence, or introducing a change in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium by gene recombination to make it close to a consensus sequence, to obtain mutants of the coryneform amino acid- or nucleic acid-producing microorganism, culturing the mutants and selecting a mutant capable of producing the intended amino acid or nucleic acid in a large amount. This method allows the construction of a mutant capable of enriching or controlling the expression of an intended gene without using a plasmid and to promote production of amino acids in a high yield by recombination or mutation.
摘要翻译：制备具有改善的氨基酸或核酸生产力的棒状细菌的方法包括以下步骤：在棒状细菌染色体上的氨基酸或核酸 - 生物合成基因的启动子序列中引入突变，使其接近共有序列或通过基因重组在棒状细菌的染色体上引入氨基酸或核酸 - 生物合成基因的启动子序列的变化使其接近共有序列，以获得棒状细胞氨基酸或核酸的突变体培养微生物，培养突变体并选择能够大量产生所需氨基酸或核酸的突变体。该方法允许构建能够富集或控制预期基因的表达而不使用质粒的突变体，并通过重组或突变促进高产量的氨基酸的产生。

4. 发明授权

US06303383B1 Temperature sensitive plasmid for coryneform bacteria 有权
标题翻译：棒状细菌的温度敏感质粒
公开(公告)号：US06303383B1
公开(公告)日：2001-10-16
申请号：US09521668
申请日：2000-03-08
申请人： Jun Nakamura , Sohei Kanno , Eiichiro Kimura , Kazuhiko Matsui , Tsuyoshi Nakamatsu
发明人： Jun Nakamura , Sohei Kanno , Eiichiro Kimura , Kazuhiko Matsui , Tsuyoshi Nakamatsu
IPC分类号： C12N1574
CPC分类号： C12N15/77
摘要： A coryneform bacterium in which a DNA fragment is incorporated into its chromosome is prepared by (a) obtaining a recombinant plasmid through ligation of a DNA fragment having a sequence homologous to a gene present on a chromosome of a coryneform bacterium to a plasmid that has a wild-type replication control region segment of a particular nucleotide sequence including a mutation and is autonomously replicable in a coryneform bacterium cell at a culture temperature lower than 31° C. but not autonomously replicable in the cell at a temperature of 31° C. or higher, (b) introducing the recombinant plasmid into the coryneform bacterium cell, (c) culturing the bacterium at a temperature of 31° C. or higher, (d) causing homologous recombination between the DNA fragment and the gene present on the chromosome of the coryneform bacterium and having a sequence homologous to the DNA fragment, and (e) selecting a coryneform bacterium in which the DNA fragment is incorporated into its chromosome. According to the present invention, there is provided a method for efficiently modifying genetic traits of a host in a short period of time by obtaining a temperature sensitive plasmid from a plasmid not exhibiting homology with already reported temperature sensitive plasmids or not exhibiting incompatibility therewith.
摘要翻译：通过（a）通过将具有与存在于棒状杆菌型细菌的染色体上的基因同源的序列的DNA片段连接到具有下列基因的质粒来获得重组质粒来制备将DNA片段并入其染色体中的棒状细菌：包含突变的特定核苷酸序列的野生型复制控制区段，并且在棒状细菌细胞中，在低于31℃的培养温度下是可自主复制的，但在31℃的温度下不能在细胞中自主复制，或（b）将重组质粒引入棒状细菌细胞中，（c）在31℃或更高的温度下培养细菌，（d）使DNA片段与存在于染色体上的基因之间的同源重组所述棒状细菌具有与所述DNA片段同源的序列，和（e）选择其中所述DNA片段并入其染色体中的棒状细菌 e。根据本发明，提供了一种通过从未报道的温度敏感性质粒不显示同源性的质粒或不表现出不相容性的质粒获得温度敏感性质粒来在短时间内高效地修饰宿主遗传性状的方法。

5. 发明授权

US5977331A .alpha.-Ketoglutarate dehydrogenase gene 失效
标题翻译： α-酮戊二酸脱氢酶基因
公开(公告)号：US5977331A
公开(公告)日：1999-11-02
申请号：US750152
申请日：1996-12-12
申请人： Yoko Asakura , Yoshihiro Usuda , Nobuharu Tsujimoto , Eiichiro Kimura , Chizu Abe , Yoshio Kawahara , Tsuyoshi Nakamatsu , Osamu Kurahashi
发明人： Yoko Asakura , Yoshihiro Usuda , Nobuharu Tsujimoto , Eiichiro Kimura , Chizu Abe , Yoshio Kawahara , Tsuyoshi Nakamatsu , Osamu Kurahashi
IPC分类号： C12N1/21 , C12N9/02 , C12N15/53 , C12P13/08 , C12P13/14 , C07H21/02 , C12N1/20 , C12P13/04
CPC分类号： C12N9/0008 , C12P13/08 , C12P13/14
摘要： Disclosed are coryneform L-glutamic acid-producing bacteria deficient in .alpha.-ketoglutarate dehydrogenase, a method of producing L-glutamic acid by using the bacteria, a gene coding for an enzyme having .alpha.-KGDH activity originating from coryneform L-glutamic acid-producing bacteria, recombinant DNA containing the gene, coryneform bacteria harboring the recombinant DNA, and a method of producing L-lysine by using bacteria harboring the recombinant DNA and having L-lysine productivity.
摘要翻译： PCT No.PCT / JP95 / 01131 Sec。 371日期：1996年12月12日 102（e）日期1996年12月12日PCT提交1995年6月7日PCT公布。 WO95 / 34672 PCT公开号日期1995年12月21日公开是α-酮戊二酸脱氢酶缺乏的棒状L-谷氨酸生产菌，使用细菌生产L-谷氨酸的方法，编码具有源自棒状杆菌L的α-KGDH活性的酶的基因谷氨酸生产菌，含有该基因的重组DNA，携带重组DNA的棒状细菌，以及通过使用含有重组DNA并具有L-赖氨酸生产力的细菌产生L-赖氨酸的方法。

6. 发明授权

US07183403B2 Genes for heat resistant enzymes of amino acid biosynthetic pathway derived from thermophilic coryneform bacteria 有权
标题翻译：来自嗜热棒状杆菌的氨基酸生物合成途径耐热酶基因
公开(公告)号：US07183403B2
公开(公告)日：2007-02-27
申请号：US11073560
申请日：2005-03-08
申请人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
发明人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
IPC分类号： C07H21/04 , C12P21/06
CPC分类号： C12Y108/01004 , C07K14/34 , C12N9/0006 , C12N9/0008 , C12N9/0051 , C12N9/1205 , C12N9/2408 , C12N9/88 , C12Y101/01041 , C12Y102/01051 , C12Y102/04002 , C12Y104/01004 , C12Y203/03001 , C12Y207/01011 , C12Y302/01026 , C12Y401/01031 , C12Y401/03001 , C12Y402/01003 , C12Y604/01001 , C12Y604/01002
摘要： A plurality of primer sets are designed based on a region where conservation at the amino acid level is observed among various microorganisms for known gene sequences corresponding to a gene coding for an enzyme of the L-amino acid biosynthetic pathway derived from Corynebacterium thermoaminogenes, preferably an enzyme that functions at a higher temperature compared with that of Corynebacterium glutamicum. PCR is performed by using the primers and chromosomal DNA of Corynebacterium thermoaminogenes as a template. The primers with which an amplification fragment has been obtained are used as primers for screening to select a clone containing a target DNA fragment from a plasmid library of chromosomal DNA of Corynebacterium thermoaminogenes.
摘要翻译：基于对应于编码源自热棒状棒杆菌的L-氨基酸生物合成途径的酶的基因的已知基因序列的各种微生物，观察到在氨基酸水平上保持的区域的多个引物组，优选为与谷氨酸棒杆菌相比在更高的温度下起作用的酶。通过使用热源性棒状杆菌的引物和染色体DNA作为模板进行PCR。使用已经获得扩增片段的引物作为引物用于筛选，从嗜热氨基酸棒杆菌的染色体DNA的质粒文库中选择含有靶DNA片段的克隆。

7. 发明授权

US07125977B2 Genes for heat resistant enzymes of amino acid biosynthetic pathway derived from thermophilic coryneform bacteria 有权
标题翻译：来自嗜热棒状杆菌的氨基酸生物合成途径耐热酶基因
公开(公告)号：US07125977B2
公开(公告)日：2006-10-24
申请号：US11073550
申请日：2005-03-08
申请人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
发明人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
IPC分类号： C07H21/04
CPC分类号： C12Y108/01004 , C07K14/34 , C12N9/0006 , C12N9/0008 , C12N9/0051 , C12N9/1205 , C12N9/2408 , C12N9/88 , C12Y101/01041 , C12Y102/01051 , C12Y102/04002 , C12Y104/01004 , C12Y203/03001 , C12Y207/01011 , C12Y302/01026 , C12Y401/01031 , C12Y401/03001 , C12Y402/01003 , C12Y604/01001 , C12Y604/01002
摘要： A plurality of primer sets are designed based on a region where conservation at the amino acid level is observed among various microorganisms for known gene sequences corresponding to a gene coding for an enzyme of the L-amino acid biosynthetic pathway derived from Corynebacterium thermoaminogenes, preferably an enzyme that functions at a higher temperature compared with that of Corynebacterium glutamicum. PCR is performed by using the primers and chromosomal DNA of Corynebacterium thermoaminogenes as a template. The primers with which an amplification fragment has been obtained are used as primers for screening to select a clone containing a target DNA fragment from a plasmid library of chromosomal DNA of Corynebacterium thermoaminogenes.
摘要翻译：基于对应于编码源自热棒状棒杆菌的L-氨基酸生物合成途径的酶的基因的已知基因序列的各种微生物，观察到在氨基酸水平上保持的区域的多个引物组，优选为与谷氨酸棒杆菌相比在更高的温度下起作用的酶。通过使用热源性棒状杆菌的引物和染色体DNA作为模板进行PCR。使用已经获得扩增片段的引物作为引物用于筛选，从嗜热氨基酸棒杆菌的染色体DNA的质粒文库中选择含有靶DNA片段的克隆。

8. 发明授权

US06995250B1 Thermophilic amino acid biosynthesis system enzyme gene of thermotolerant coryneform bacterium 失效
标题翻译：耐热棒状杆菌型嗜热氨基酸生物合成酶基因
公开(公告)号：US06995250B1
公开(公告)日：2006-02-07
申请号：US10089057
申请日：2000-10-04
申请人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
发明人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
IPC分类号： C07H21/02 , C07H21/04 , C12P21/06
CPC分类号： C12Y108/01004 , C07K14/34 , C12N9/0006 , C12N9/0008 , C12N9/0051 , C12N9/1205 , C12N9/2408 , C12N9/88 , C12Y101/01041 , C12Y102/01051 , C12Y102/04002 , C12Y104/01004 , C12Y203/03001 , C12Y207/01011 , C12Y302/01026 , C12Y401/01031 , C12Y401/03001 , C12Y402/01003 , C12Y604/01001 , C12Y604/01002
摘要： A plurality of primer sets are designed based on a region where conservation at the amino acid level is observed among various microorganisms for known gene sequences corresponding to a gene coding for an enzyme of the L-amino acid biosynthetic pathway derived from Corynebacterium thermoaminogenes, preferably an enzyme that functions at a higher temperature compared with that of Corynebacterium glutamicum. PCR is performed by using the primers and chromosomal DNA of Corynebacterium thermoaminogenes as a template. The primers with which an amplification fragment has been obtained are used as primers for screening to select a clone containing a target DNA fragment from a plasmid library of chromosomal DNA of Corynebacterium thermoaminogenes.
摘要翻译：基于对应于编码源自热棒状棒杆菌的L-氨基酸生物合成途径的酶的基因的已知基因序列的各种微生物，观察到在氨基酸水平上保持的区域的多个引物组，优选为与谷氨酸棒杆菌相比在更高的温度下起作用的酶。通过使用热源性棒状杆菌的引物和染色体DNA作为模板进行PCR。使用已经获得扩增片段的引物作为引物用于筛选，从嗜热氨基酸棒杆菌的染色体DNA的质粒文库中选择含有靶DNA片段的克隆。

9. 发明申请

US20050239176A1 Genes for heat resistant enzymes of amino acid biosynthetic pathway derived from thermophilic coryneform bacteria 有权
标题翻译：来自嗜热棒状杆菌的氨基酸生物合成途径耐热酶基因
公开(公告)号：US20050239176A1
公开(公告)日：2005-10-27
申请号：US11073560
申请日：2005-03-08
申请人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
发明人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
IPC分类号： C07K14/34 , C12N9/02 , C12N9/04 , C12N9/12 , C12N9/26 , C12N9/88 , C12N15/31 , C12N15/53 , C12N15/54 , C12N15/56 , C12N15/60 , C12P13/04 , C07H21/04 , C12N9/10 , C12N15/74 , C12P13/08
CPC分类号： C12Y108/01004 , C07K14/34 , C12N9/0006 , C12N9/0008 , C12N9/0051 , C12N9/1205 , C12N9/2408 , C12N9/88 , C12Y101/01041 , C12Y102/01051 , C12Y102/04002 , C12Y104/01004 , C12Y203/03001 , C12Y207/01011 , C12Y302/01026 , C12Y401/01031 , C12Y401/03001 , C12Y402/01003 , C12Y604/01001 , C12Y604/01002
摘要： A plurality of primer sets are designed based on a region where conservation at the amino acid level is observed among various microorganisms for known gene sequences corresponding to a gene coding for an enzyme of the L-amino acid biosynthetic pathway derived from Corynebacterium thermoaminogenes, preferably an enzyme that functions at a higher temperature compared with that of Corynebacterium glutamicum. PCR is performed by using the primers and chromosomal DNA of Corynebacterium thermoaminogenes as a template. The primers with which an amplification fragment has been obtained are used as primers for screening to select a clone containing a target DNA fragment from a plasmid library of chromosomal DNA of Corynebacterium thermoaminogenes.
摘要翻译：基于对应于编码源自热棒状棒杆菌的L-氨基酸生物合成途径的酶的基因的已知基因序列的各种微生物，观察到在氨基酸水平上保持的区域的多个引物组，优选为与谷氨酸棒杆菌相比在更高的温度下起作用的酶。通过使用热源性棒状杆菌的引物和染色体DNA作为模板进行PCR。使用已经获得扩增片段的引物作为引物用于筛选，从嗜热氨基酸棒杆菌的染色体DNA的质粒文库中选择含有靶DNA片段的克隆。

10. 发明申请

US20050176127A1 Genes for heat resistant enzymes of amino acid biosynthetic pathway derived from thermophilic coryneform bacteria 有权
公开(公告)号：US20050176127A1
公开(公告)日：2005-08-11
申请号：US11073550
申请日：2005-03-08
申请人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
发明人： Seiko Hirano , Eiichiro Kimura , Tsuyoshi Osumi , Kazuhiko Matsui , Yoshio Kawahara , Gen Nonaka , Yumi Matsuzaki , Naoki Akiyoshi , Kanae Nakamura , Osamu Kurahashi , Tsuyoshi Nakamatsu , Shinichi Sugimoto
IPC分类号： C07K14/34 , C12N9/02 , C12N9/04 , C12N9/12 , C12N9/26 , C12N9/88 , C12N15/31 , C12N15/53 , C12N15/54 , C12N15/56 , C12N15/60 , C12N1/21 , C12N15/74 , C12P13/08
CPC分类号： C12Y108/01004 , C07K14/34 , C12N9/0006 , C12N9/0008 , C12N9/0051 , C12N9/1205 , C12N9/2408 , C12N9/88 , C12Y101/01041 , C12Y102/01051 , C12Y102/04002 , C12Y104/01004 , C12Y203/03001 , C12Y207/01011 , C12Y302/01026 , C12Y401/01031 , C12Y401/03001 , C12Y402/01003 , C12Y604/01001 , C12Y604/01002
摘要： A plurality of primer sets are designed based on a region where conservation at the amino acid level is observed among various microorganisms for known gene sequences corresponding to a gene coding for an enzyme of the L-amino acid biosynthetic pathway derived from Corynebacterium thermoaminogenes, preferably an enzyme that functions at a higher temperature compared with that of Corynebacterium glutamicum. PCR is performed by using the primers and chromosomal DNA of Corynebacterium thermoaminogenes as a template. The primers with which an amplification fragment has been obtained are used as primers for screening to select a clone containing a target DNA fragment from a plasmid library of chromosomal DNA of Corynebacterium thermoaminogenes.

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