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Construction of novel strains containing minimizing genome by tn5-coupled cre/loxp excision system
Construction of novel strains containing minimizing genome by tn5-coupled cre/loxp excision system
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机译:用tn5偶联的cre / loxp切除系统构建含有最小化基因组的新菌株
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摘要
Disclosed is a method for developing novel strains deleted specific chromosome sites, using transposon and Cre/loxP site-specific recombination by Cre expression vector, wherein the transposon comprises a selectable marker and loxP site. The method comprises the steps of: (1) preparing a transposon comprising a selectable marker and loxP site; (2) inserting the transposon into an optional position of microbial chromosome, and determining the inserted site; (3) integrating two transposons comprising a different selectable marker to one chromosome; (4) deleting a chromosomal site between the two lox sites by introducing a Cre expression vector into the chromosome of step (3); and (5) repeating steps (3 and 4) for the mutant deleted a part of chromosome, to shorten the chromosome of mutant gradually.
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