Abstract
The invention relates to a gene editing system of Escherichia coliincluding a helper plasmid and a donor plasmid. The helper plasmid includes a transposase complex expression cassette, a Cas12k expression cassette, a first sgRNA cassette, a first antibiotic resistance gene and first replication origin. The donor plasmid includes a left end sequence of the ShCAST transposon, an exogenous gene expression cassette, a right end sequence of the ShCAST transposon, a second sgRNA cassette, a second antibiotic resistance gene and a second replication origin. The gene editing system of Escherichia colican easily and effectively integrate large fragments of DNA sequences into target chromosome without causing double-strand breaks in the genome of Escherichia coli, so as to regulate the metabolic pathways of Escherichia colistrains useful in different biotechnologies.