摘要
Recombineering is a valuable technique for generating recombinant DNA in vivo,primarily in bacterial cells,and is based on homologous recombination using phage-encoded homologous recombinases,such as Red βγ from the lambda phage and RecET from the Rac prophage.The recombineering technique can efficiently mediate homol-ogous 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 Escherichia coli 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.
基金
supported by the National Key R&D Program of China(2019YFA0904000)
the National Natural Science Foundation of China(31570094,81502962,32170038,32270088)
the Taishan Scholar Pro-gram of Shandong Province,the Shandong Provincial Natural Science Foundation of China(ZR2020MC015,ZR2022MC142)
the Funda-mental Research Funds of Shandong University(2018GN021).