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

作　　者：司微[1] 刘慧芳[1] 王春来[1] 彭玮[1] 赫明雷[1] 杜艳芬[1] 赵海玲[1] 王聃[1] 杨金国[1] 林靖凯[1] 倪洪波[1] 刘思国[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室动物细菌病研究室,哈尔滨150001

出　　处：《黑龙江畜牧兽医》2011年第8期28-30,共3页Heilongjiang Animal Science And veterinary Medicine

基　　金：中国农业科学院哈尔滨兽医研究所中央级公益性科研院所基本科研业务费专项(2008-14)

摘　　要：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.

关 键 词：RED重组系统 ClpP基因 基因敲除 

分 类 号：S852.65[农业科学—基础兽医学]