| Abstract |
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Replacement recombination in Coryneform bacteria: high efficiency integration requirement for non-methylated plasmid DNA. |
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| 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. Integratetive plasmid DNA containing an invitro disrupted copy of the Brevibacterium flavum thrB gene was introduced into B. flavum at an efficiency of 2.4 x 102 integrants/microgram. Integration occurred by homologous recombination either via a Campbell-like mechanism or via a double crossover event. Integrants were stable for over 32 generations. |