Department of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657 5801, Japan.
School of Computing, Newcastle University, 1 Science Square, Science Central, Newcastle upon Tyne, NE4 5TG, UK.
Microb Cell Fact. 2022 Mar 8;21(1):34. doi: 10.1186/s12934-022-01759-8.
Geobacillus kaustophilus is a thermophilic Gram-positive bacterium. Methods for its transformation are still under development. Earlier studies have demonstrated that pLS20catΔoriT mobilized the resident mobile plasmids from Bacillus subtilis to G. kaustophilus and transferred long segments of chromosome from one cell to another between B. subtilis.
In this study, we applied mobilization of the B. subtilis chromosome mediated by pLS20catΔoriT to transform G. kaustophilus. We constructed a gene cassette to be integrated into G. kaustophilus and designed it within the B. subtilis chromosome. The pLS20catΔoriT-mediated conjugation successfully transferred the gene cassette from the B. subtilis chromosome into the G. kaustophilus allowing for the desired genetic transformation.
This transformation approach described here will provide a new tool to facilitate the flexible genetic manipulation of G. kaustophilus.
解淀粉芽孢杆菌是一种嗜热革兰氏阳性细菌。其转化方法仍在开发中。早期的研究表明，pLS20catΔoriT 可将枯草芽孢杆菌中的常驻移动质粒转移到解淀粉芽孢杆菌中，并在枯草芽孢杆菌之间将染色体的长片段从一个细胞转移到另一个细胞。
在本研究中，我们应用 pLS20catΔoriT 介导的枯草芽孢杆菌染色体的动员来转化解淀粉芽孢杆菌。我们构建了一个基因盒，将其整合到解淀粉芽孢杆菌中，并在枯草芽孢杆菌染色体内进行设计。pLS20catΔoriT 介导的共轭成功地将基因盒从枯草芽孢杆菌染色体转移到解淀粉芽孢杆菌中，从而实现了所需的遗传转化。
这里描述的这种转化方法将为解淀粉芽孢杆菌的灵活遗传操作提供新的工具。
