REPLACEMENT RECOMBINATION IN CORYNEFORM BACTERIA - HIGH-EFFICIENCY INTEGRATION REQUIREMENT FOR NONMETHYLATED PLASMID DNA

被引:33
作者
VERTES, AA [1 ]
HATAKEYAMA, K [1 ]
INUI, M [1 ]
KOBAYASHI, M [1 ]
KURUSU, Y [1 ]
YUKAWA, H [1 ]
机构
[1] MITSUBISHI PETROCHEM CO LTD, TSUKUBA RES CTR, INASHIKI, IBARAKI 30003, JAPAN
关键词
D O I
10.1271/bbb.57.2036
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have developed a versatile system that permits efficient manipulation of coryneform bacteria by gene disruption and replacement. Circumventing the methyl-specific restriction system in certain of these bacteria by using non-methylated plasmid DNA was necessary. Integrative plasmid DNA containing an in vitro disrupted copy of the Brevibacterium flavum thrB gene was introduced into B. flavum at an efficiency of 2.4 x 10(2) integrants/mug. Integration occurred by homologous recombination either via a Campbell-like mechanism or via a double crossover event. Integrants were stable for over 32 generations.
引用
收藏
页码:2036 / 2038
页数:3
相关论文
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