Genome minimization by tn5-coupled cre/loxP excision system

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.