Liu Qingshu, Li Ruijuan, Shi Hongbo, Yang Runyu, Shen Qiyao, Cui Qingwen, Wang Xiuling, Li Aiying, Zhang Youming, Fu Jun
State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China.
Hunan Institute of Microbiology, Xinkaipu Lu 18, Tianxin District, Changsha 410009, China.
Eng Microbiol. 2023 Jun 21;3(3):100099. doi: 10.1016/j.engmic.2023.100099. eCollection 2023 Sep.
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 have low efficiency. In this work, our goal was to develop a simple recombineering system for . We showed that genome editing can be achieved in 1A751 through co-expression of YqaJ/YqaK, a native phage recombinase pair found in 168, and the competence master regulator ComK using a double-stranded DNA substrate with short homology 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 1A751. As far as we know, this is the first recombineering system using the native phage recombinase pair YqaJ/YqaK in . In conclusion, this new recombineering system provides a simple and fast tool for genetic manipulation of , and it will promote studies of genome function, construction of production strains, and genome mining in .
在基础研究和应用研究中发挥着重要作用，并且它已被广泛用作细胞工厂，用于生产酶、抗菌材料以及农业、医药和工业用化学品。然而，用于[具体对象]的基因操作工具效率较低。在这项工作中，我们的目标是为[具体对象]开发一种简单的重组工程系统。我们表明，通过共表达YqaJ/YqaK（在168中发现的一对天然噬菌体重组酶）和感受态主调节因子ComK，利用具有短同源臂（100 bp）且5'-端有硫代磷酸酯修饰的双链DNA底物，可以在1A751中实现基因组编辑。使用这种新的重组工程系统在1A751中实现了高效的基因敲除和大DNA插入。据我们所知，这是第一个在[具体对象]中使用天然噬菌体重组酶对YqaJ/YqaK的重组工程系统。总之，这种新的重组工程系统为[具体对象]的基因操作提供了一种简单快速的工具，并且它将促进[具体对象]中的基因组功能研究、生产菌株构建和基因组挖掘。
