A recombineering system for Bacillus subtilis based on the native phage recombinase pair YqaJ/YqaK

查看全文 作　　者：Qingshu [1,2]Liu;Ruijuan [1]Li;Hongbo [1]Shi;Runyu [1]Yang;Qiyao [1]Shen;Qingwen [1]Cui;Xiuling [1]Wang;Aiying [1]Li;Youming [1]Zhang;Jun [1]Fu 机构地区：[1]State Key Laboratory of Microbial Technology,Shandong University,Qingdao 266237,China;[2]Hunan Institute of Microbiology,Xinkaipu Lu 18,Tianxin District,Changsha 410009,China 出　　处：《Engineering Microbiology》索引2023年第3卷第3期,共9页 基　　金：supported by the National Key R&D Program of China(2019YFA0904000);the National Natural Science Foundation of China(31570094,81502962,32270088);the 111 Project(B16030);the Shan-dong Provincial Natural Science Foundation of China(ZR2020MC015,ZR2018ZC2261);the Taishan Scholar Program of Shandong Province;the Fundamental Research Funds of Shandong University(2018GN021);the Open Project Program of the State Key Laboratory of Bio-based Material and Green Papermaking(KF201825);the Science and Technology Project of Hunan Province(2021NK1040);Natural Science Foundation of Changsha(KQ2208130). 摘　　要：Bacillus subtilis plays an important role in fundamental and applied research,and it has been widely used as a cell factory for the production of enzymes,antimicrobial materials,and chemicals for agriculture,medicine,and industry.However,genetic manipulation tools for B.subtilis have low efficiency.In this work,our goal was to develop a simple recombineering system for B.subtilis.We showed that genome editing can be achieved in B.subtiliis 1A751 through co-expression of YqaJ/YqaK,a native phage recombinase pair found in B.sub-tilis 168,and the competence master regulator ComK using a double-stranded DNA substrate with short ho-mology arms(100 bp)and a phosphorothioate modification at the 5′-end.Efficient gene knockouts and large DNA insertions were achieved using this new recombineering system in B.subtilis 1A751.As far as we know,this is the first recombineering system using the native phage recombinase pair YqaJ/YqaK in B.subtilis.In conclusion,this new recombineering system provides a simple and fast tool for genetic manipulation of B.sub-tilis,and it will promote studies of genome function,construction of production strains,and genome mining in B.subtilis. 关 键 词：Bacillus subtilis RECOMBINEERING YqaJK system Genome editing