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Background  The conjugative plasmid, pLS20, isolated from  Bacillus subtilis  natto, has an outstanding capacity for rapid self-transfer. In addition, it can function as a helper plasmid, mediating the mobilization of an independently replicating co-resident plasmid.    Results  In this study, the  oriT  sequence of pLS20cat ( oriT  LS20 ) was eliminated to obtain the plasmid, pLS20catΔ oriT . This resulted in the complete loss of the conjugative transfer of the plasmid but still allowed it to mobilize a co-resident mobilizable plasmid. Moreover, pLS20catΔ oriT  was able to mobilize longer DNA segments, up to 113 kb of chromosomal DNA containing  oriT  LS20 , after mixing the liquid cultures of the donor and recipient for only 15 min.    Conclusions  The chromosomal DNA mobilization mediated by pLS20catΔ oriT  will allow us to develop a novel genetic tool for the rapid, easy, and repetitive mobilization of longer DNA segments into a recipient chromosome.

microbial cell factories

Rapid conjugative mobilization of a 100 kb segment of Bacillus subtilis chromosomal DNA is mediated by a helper plasmid with no ability for self-transfer