摘要
通过电转化将一段PCR产物引入宿主菌BW25113/pIJ790细胞内,PCR产物两端有与染色体同源的30个核苷酸序列,中间是抗生素抗性基因。pIJ790上编码λ噬菌体的3个重组蛋白:Exo、Bet和Gam组成Red重组系统,可实现线性片段的一步法高效重组,以PCR产物中的抗生素抗性基因取代靶基因。通过该方法得到了大肠埃希菌的外排泵基因acrAB敲除突变株,该菌株在抗菌物质的抗菌机制研究方面能发挥一定的作用。
A PCR fragment including 30nt of flanking DNA homologous to the chromosome in each end and a antibiotic-resistant gene in the middle is introduced into the host cell of BW25113/ pIJ790. The three proteins of h phage---Exo,Bet and Gam, expressed by pIJ790, composing the Red recombination system has the advantage of high one-step recombination rate of linear fragment. The antibiotic-resistant gene replaces the target gene as a result. This research obtaines a derivative of E. coli whose efflux pump gene aerAB is deleted. The mutant strain will help to research of mechanism of antimierobial materials.
出处
《江汉大学学报（自然科学版）》
2012年第5期76-79,共4页
Journal of Jianghan University：Natural Science Edition
基金
武汉市科技局青年科技晨光计划项目(201271031394)
环境化学与生态毒理学国家重点实验室开放基金课题(KF2011-06)
