Cloning of multiple genes involved with cobalamin (Vitamin B12) biosynthesis in Bacillus megaterium.

摘要: An effective shotgun cloning procedure was developed for Bacillus megaterium by amplifying gene libraries in subtilis. This technique useful isolating at least 11 genes from B. which are involved with cobalamin (vitamin B12) biosynthesis. Amplified plasmid banks were transformed into protoplasts of both a series Cob mutants blocked before the biosynthesis cobinamide and Cbl conversion cobalamin. Amplification overcame barriers inherent relatively low protoplast transformation frequency megaterium. A family plasmids isolated complementation seven different mutants. Each capable complementing or mutant each one mutants; many mutation carried genetic activity other mutations. By these criteria, four groups resolved. At six on fragment DNA approximately 2.7 kilobase pairs length; located two groups. The physical data permitted an ordering within several presence synthesis resulted restoration