发明授权
- 专利标题: Temperature sensitive plasmid for coryneform bacteria
- 专利标题(中): 棒状细菌的温度敏感质粒
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申请号: US09521668申请日: 2000-03-08
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公开(公告)号: US06303383B1公开(公告)日: 2001-10-16
- 发明人: Jun Nakamura , Sohei Kanno , Eiichiro Kimura , Kazuhiko Matsui , Tsuyoshi Nakamatsu
- 申请人: Jun Nakamura , Sohei Kanno , Eiichiro Kimura , Kazuhiko Matsui , Tsuyoshi Nakamatsu
- 优先权: JP11-069896 19990316
- 主分类号: C12N1574
- IPC分类号: C12N1574
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.