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

Author: Jing WANG ¹ ; Ruichen LV ² ; Yanping HAN ² ; Ruifu YANG ²
Author Information

1. College of Animal Science and Technology, Hebei North University, Zhangjiakou 075131, Hebei, China.
2. State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.
Publication Type:Journal Article
Keywords: Escherichia coli; endonucleaseⅠ-SceⅠ; fusion PCR; hfq; rne-710; two-plasmid scarless genetic modification
From: Chinese Journal of Biotechnology 2018;34(4):602-612
CountryChina
Language:Chinese
Abstract: Gene modification is an important technique to understand gene function. We firstly constructed Δhfq::Spe and Δrne-710::Spe 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.