Li Jie, Wang Baiyang, Yang Qing, Si Han, Zhao Yuting, Zheng Yanli, Peng Wenfang
State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Engineering Research Center for Bio-enzyme Catalysis, Environmental Microbial Technology Center of Hubei Province, School of Life Sciences, Hubei University, Wuhan, China.
College of Life Science and Technology, Wuhan Polytechnic University, Wuhan, China.
Front Microbiol. 2022 Mar 3;13:848964. doi: 10.3389/fmicb.2022.848964. eCollection 2022.
species are emerging as important microorganisms of global concern with unique and increasingly significant ecological roles and represent a prominent source of bioactive natural products, but genetic engineering of these organisms for biotechnological applications is greatly hindered due to the limitation of efficient genetic manipulation tools. In this regard, we report here the establishment of an efficient genetic manipulation system for a newly isolated strain, Shahu, based on plasmid conjugal transfer from to . Conjugants were yielded upon determining the optimal ratio between the donor and recipient cells, and designed genome modifications were efficiently accomplished, including exogenous gene integration based on an integrative plasmid and chromosomal stretch removal by homologous recombination using a suicidal non-replicating vector. Collectively, this work has made the Shahu accessible for genetic engineering, and provided an important reference for developing genetic manipulation methods in other rare actinomycetes.
物种正成为全球关注的重要微生物，具有独特且日益重要的生态作用，是生物活性天然产物的重要来源，但由于高效基因操作工具的限制，这些生物在生物技术应用中的基因工程受到极大阻碍。在这方面，我们在此报告基于从 到 的质粒接合转移，为新分离的菌株沙湖建立了一种高效的基因操作系统。在确定供体和受体细胞之间的最佳比例后产生了接合子，并有效地完成了设计的基因组修饰，包括基于整合质粒的外源基因整合和使用自杀性非复制载体通过同源重组去除染色体片段。总的来说，这项工作使沙湖能够进行基因工程操作，并为开发其他稀有放线菌的基因操作方法提供了重要参考。
