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

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 s...

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Detalles Bibliográficos
Autores: Miyano, Megumi, Tanaka, Kosei, Ishikawa, Shu, Takenaka, Shinji, Miguel-Arribas, Andrés, Meijer, Wilfried J. J., Yoshida, Ken-ichi
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2018
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/181643
Acceso en línea:http://hdl.handle.net/10261/181643
Access Level:acceso abierto
Palabra clave:Gram positive
Conjugation
Bacillus subtilis
Plasmid transfer
Descripción
Sumario: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 ) was eliminated to obtain the plasmid, pLS20catoriT. 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, pLS20catoriT was able to mobilize longer DNA segments, up to 113kb of chromosomal DNA containing oriT , after mixing the liquid cultures of the donor and recipient for only 15min. Conclusions: The chromosomal DNA mobilization mediated by pLS20catoriT will allow us to develop a novel genetic tool for the rapid, easy, and repetitive mobilization of longer DNA segments into a recipient chromosome.