[Two-plasmid scarless genetic modification in Escherichia coli hfq and rne-710].

ABSTRACT

Gene modification is an important technique to understand gene function. We firstly constructed ΔhfqSpe and Δrne-710Spe mutant strains of Escherichia coli MG1655. The fragment lacking of hfq and rne-710 was ligated to the auxiliary plasmid and separately replace the spectinomycin box by homologous recombinase system to obtain the Δhfq and Δrne-710 mutant strains. The combination of two-plasmid scarless genetic modification and fusion PCR led to a new way for the long DNA fragment gene deletions.