A novel method for transforming Geobacillus kaustophilus with a chromosomal segment of Bacillus subtilis transferred via pLS20-dependent conjugation

Abstract

Background 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. Results 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. Conclusions This transformation approach described here will provide a new tool to facilitate the flexible genetic manipulation of G. kaustophilus.

A conjugative plasmid exploits flagella rotation as a cue to facilitate its transfer

“…The 65 Kb conjugative plasmid pLS20, initially isolated from the Gram-positive Bacillus subtilis (B. subtilis) natto strain 14 , possesses transmission capacity to other Bacilli, including the pathogenic species B. anthracis and B. cereus, highlighting its potential to serve as a vehicle for interspecies genetic exchange [15][16][17] . In fact, pLS20 serves as the archetype plasmid to a family of at least 35 members, dispersed across various Bacillus species residing in diverse ecological niches 18 .…”