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1.发明授权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
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。
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2.发明申请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
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。
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3.发明授权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
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。
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公开(公告)号：US06303383B1
公开(公告)日：2001-10-16
申请号：US09521668
申请日：2000-03-08
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。
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