Red同源重组技术发展迅速,已经广泛应用于大肠杆菌基因的敲除、插入与替换。与传统的DNA有痕重组技术相比,基于Red重组原理的DNA无痕重组技术,能够更为精确、快速、高效地修饰大肠杆菌基因组中的目标基因,且在基因组中不残留任何外源片段,因此不会影响后续的基因操作与基因表达。从Red同源重组的原理出发,简要综述了近年来在大肠杆菌中广泛使用的无痕重组技术的原理及操作策略,并对比分析了各种方法的优势与不足;同时,还介绍了DNA无痕重组技术在大肠杆菌基因修饰中的应用情况。
Red recombination technology is developing rapidly,and has been widely used in Escherichia coli gene modification,such as deletions,insertions and substitutions. Compared with the traditional scarred recombination,the scarless recombination technology based on Red recombination is able to make more precise,rapid and efficient modification of targeted genes without the introduction of any exogenous sequences in the genome. From the basic mechanism of Red recombination,the principles and operation strategies about scarless recombination for Escherichia coli in recent years were reviewed,and also the advantages and disadvantages were analyzed. Finally,the applications of scarless recombination technology were introduced.