利用Red重组系统敲除大肠杆菌菌株ClpP基因方法的研究

The research on the deletion of ClpP gene in chromosome of E. coli by Red recombination system

为了研究ClpP基因的功能,试验利用Red系统的重组功能,通过PCR扩增出两端与大肠杆菌菌株ClpP基因上、下游序列同源,中间为氯霉素抗性基因cat的DNA片段,电击转化含质粒pKD46的大肠杆菌菌株DH091,筛选替换ClpP基因的阳性转化子,再导入质粒pCP20去除氯霉素抗性基因。结果表明:成功敲除大肠杆菌菌株ClpP基因。

With the help of Red recombination system, the chloramphenicol resistance gene flanked by homologues of the ClpP gene was amplified by PCR. The PCR products were electro - transformed into E. coli DH091 strain with pKD46. After ClpP gene was replaced by the ehloramphenicol gene, the resistance gene was then eliminated by using pCP20, then the CIpP gene was completely deleted by this procedure. This research can provide a basis on the function of the ClpP gene.