| 利用PCR产物一步法敲除大肠埃希菌的外排泵基因acrAB |
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| 引用本文： | 纪芳,鄢志雄,李文丹,祝贺,梁勇. 利用PCR产物一步法敲除大肠埃希菌的外排泵基因acrAB[J]. 江汉大学学报(自然科学版), 2012, 40(5): 76-79 |
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| 作者姓名： | 纪芳 鄢志雄 李文丹 祝贺 梁勇 |
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| 作者单位： | 江汉大学医学院,湖北武汉,430056 |
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| 基金项目： | 武汉市科技局青年科技晨光计划项目，环境化学与生态毒理学国家重点实验室开放基金课题 |
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| 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。
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| 关 键 词： | Red重组 PCR 基因敲除 外排泵基因 |
Deletion of Efflux Pump Gene acrAB in E. coli with PCR Product One-step Method |
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| JI Fang,YAN Zhi-xiong,LI Wen-dan,ZHU He,LIANG Yong. Deletion of Efflux Pump Gene acrAB in E. coli with PCR Product One-step Method[J]. Journal of Jianghan University(Natural Sciences), 2012, 40(5): 76-79 |
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| Authors: | JI Fang YAN Zhi-xiong LI Wen-dan ZHU He LIANG Yong |
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| Affiliation: | (School of Medicine,Jianghan University,Wuhan 430056,Hubei,China) |
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| Abstract: | 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. |
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| Keywords: | Red recombination PCR gene knock-out efflux pump gene |
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