Red重组系统用于大肠杆菌基因修饰的研究进展

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。本文分别详细介绍了Red重组系统中Exo、Beta、Gam三种蛋白质的功能，Red重组系统运用在大肠杆菌基因敲除中的三种质粒及其功能，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。

Advances of Red Recombination System in Escherichia coli Gene Modification

As a new recombination system, Red recombination has been widely used in E.coli gene knockout. Comparing with Rec recombination, Red recombination has 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 in this paper. 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. We also summarized the concentration and inducing time of L-arabinose and the length of homologous arms by literature mining, and proposed the optimal parameters. 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.