Li Ruijuan, Li Aiying, Zhang Youming, Fu Jun
State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China.
Chinese Academy of Sciences (CAS) Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
Eng Microbiol. 2023 May 24;3(3):100097. doi: 10.1016/j.engmic.2023.100097. eCollection 2023 Sep.
Recombineering is a valuable technique for generating recombinant DNA , primarily in bacterial cells, and is based on homologous recombination using phage-encoded homologous recombinases, such as Redαβγ from the lambda phage and RecET from the Rac prophage. The recombineering technique can efficiently mediate homologous recombination using short homologous arms (∼50 bp) and is unlimited by the size of the DNA molecules or positions of restriction sites. In this review, we summarize characteristics of recombinases, mechanism of recombineering, and advances in recombineering for DNA manipulation in and other bacteria. Furthermore, the broad applications of recombineering for mining new bioactive microbial natural products, and for viral mutagenesis, phage genome engineering, and understanding bacterial metabolism are also reviewed.
重组工程是一种用于产生重组DNA的重要技术，主要应用于细菌细胞中，它基于使用噬菌体编码的同源重组酶进行同源重组，例如来自λ噬菌体的Redαβγ和来自Rac原噬菌体的RecET。重组工程技术可以使用短同源臂（约50bp）高效介导同源重组，并且不受DNA分子大小或限制酶切位点位置的限制。在本综述中，我们总结了重组酶的特性、重组工程的机制以及在大肠杆菌和其他细菌中进行DNA操作的重组工程进展。此外，还综述了重组工程在挖掘新型生物活性微生物天然产物、病毒诱变、噬菌体基因组工程以及理解细菌代谢方面的广泛应用。
