Red重组系统用于大肠杆菌基因修饰研究进展  

Advances of Red Recombination System in Escherichia coli Gene Modification

作　　者：张雪[1,2] 温廷益[2] 

机构地区：[1]天津科技大学生物工程学院,天津300457 [2]中国科学院微生物研究所,北京100101

出　　处：《中国生物工程杂志》2008年第12期89-93,共5页China Biotechnology

基　　金：国家"863"计划(2006AA02Z216)资助项目

摘　　要：Red重组作为一种新的重组系统已经被广泛用于大肠杆菌的基因敲除、基因替换。与传统的Rec重组相比,Red重组具有同源臂短,重组效率高等优点。分别详细介绍了Red重组系统中Exo、Beta、Gam3种蛋白质的功能,Red重组系统运用于大肠杆菌基因敲除的3种质粒及其功能,同时概括了Red重组的技术要点及技术难点,分析了文献报道的阿拉伯糖诱导浓度和诱导时间、转化后的复苏温度及时间、引物同源臂长度对于重组率的影响,总结出了Red重组的最佳条件。As a new recombination system, Red recombination has knockout. Comparing with Rec recombination, Red recombination has been widely used in E. coli gene the advantages of exploring short homologous arms and high recombination rate. The length of homologous arms usually are 36 - 60 nt. The function of Exo, Beta and Gam proteins in Red recombination, three kind of plasmids and their functions in E. coli gene knockout system were reviewed. The three kind of plasmids are plasmid which has exo, bet, gam for Red recombination, plasmid which has resistance tag and FRT site for PCR template and plasmid which has FLP recombinase for eliminating resistance tag. The concentration and inducing time of L-arabinose and the length of homologous arms by literature mining were summarized, and the optimal parameters was proposed. The optimal concentration and inducing time of L-arabinose are 100 mmoL/L and 90 min, and the optimal length of homologous arms is 56 nt.

关 键 词：RED重组 大肠杆菌 基因敲除 同源臂 L-阿拉伯糖 

分 类 号：Q819[生物学—生物工程]