Red两步同源重组法在大肠杆菌基因敲除中的应用

doi:10.16431/j.cnki.1671-7236.2017.07.005

Abstract

Abstract:

Knock-out is the most direct way to explore gene's function. The traditional method of Escherichia coli (E.coli) gene knock-out is to use its own RecA reorganization system, which relies on the specific given cleavage sites of restriction enzymes, requires long homologous arm, furthermore, the operation is complex. Modification of the genome has become simple and rapid since the advent of Red homologous recombination method. And its application in E. coli gene knock-out has been more mature. The structure and functional principle of Red homologous recombination is introduced, besides, this review expounds the operating steps and notices of the common two-step Red-mediated recombination. The Red homologous recombination technology in E.coli has made great contribution in the area of modifications of engineering strain, pathogenicity and bacterial resistance of the pathogenic bacteria. Additionally,the advantages and shortcomings of the method are set forth. According to the disadvantage that the traditional method do leave recombinase recognition site scars and has low efficiency, a sum of classical methods for scar-less gene replacement are described in the final part of the article. These approach can provide better applications for the construction of E.coli genome.

Key words: Escherichia coli; Red homologous recombination; gene knock-out; two-step

CLC Number:

Q784

LI Xin, LI Ya-xin, DAI Jian-jun. Usage of Two-step Red Homologous Recombination Method to Knockout the Gene of Escherichia coli[J]. , 2017, 44(7): 1934-1940.

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Usage of Two-step Red Homologous Recombination Method to Knockout the Gene of Escherichia coli

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