大肠杆菌基因无痕敲除技术及策略

Approaches and Strategies of Gene Scarless Knockout in the Esche richia coli Genome

基因敲除技术是大肠杆菌基因组减小和代谢途径改造的有效手段,其中基于同源重组原理的基因无痕敲除技术显现出其他技术所不具备的应用优势和发展潜力。该技术可以快速敲除大肠杆菌基因组中的目标基因,并且在基因组中不残留任何外源片段,所以不会干扰后续的基因操作。我们分类介绍了无痕敲除技术中所涉及载体的结构、功能及其相应的敲除策略,着重介绍了无痕敲除技术的原理及载体构建方法。

Gene knockout has been broadly applied in genome reduction and metabolic engineering of Escherich-ia coli. The scarless deletion technology based on homologous recombination provides clear application advantages over other knockout methods. This technology allows rapid and sequential deletion of targeted genes without intro-duction of any exogenous sequences in the genome. In this article we presented the structures and functions of dif-ferent types of vectors applied in scarless deletion, the principle and the procedure of scarless knockout technolo-gy were also described in detail.