English   español  
Por favor, use este identificador para citar o enlazar a este item: http://hdl.handle.net/10261/168855
COMPARTIR / IMPACTO:
Estadísticas
logo share SHARE   Add this article to your Mendeley library MendeleyBASE
Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL
Exportar a otros formatos:
Título

A novel method for transforming the thermophilic bacterium Geobacillus kaustophilus

AutorMiyano, Megumi; Tanaka, Kosei; Ishikawa, Shu; Mori, Kotaro; Miguel-Arribas, Andrés; Meijer, Wilfried J J; Yoshida, Ken-ichi
Fecha de publicación17-ago-2018
EditorBioMed Central
CitaciónMicrobial Cell Factories. 2018 Aug 17;17(1):127
ResumenAbstract Background Bacterial strains of the genus Geobacillus grow at high temperatures of 50–75 °C and could thus be useful for biotechnological applications. However, genetic manipulation of these species is difficult because the current techniques for transforming Geobacillus species are not efficient. In this study, we developed an easy and efficient method for transforming Geobacillus kaustophilus using the conjugative plasmid pLS20cat. Results We constructed a transformation system comprising (i) a mobilizable Bacillus subtilis–G. kaustophilus shuttle plasmid named pGK1 that carries the elements for selection and replication in Geobacillus, and (ii) a pLS20cat-harboring B. subtilis donor strain expressing the dam methylase gene of Escherichia coli and the conjugation-stimulating rapLS20 gene of pLS20cat. This system can be used to efficiently introduce pGK1 into G. kaustophilus by mobilization in a pLS20cat-dependent way. Whereas the thermostable kanamycin marker and Geobacillus replication origin of pGK1 as well as expression of dam methylase in the donor were indispensable for mobilization, ectopic expression of rapLS20 increased its efficiency. In addition, the conditions of the recipient influenced mobilization efficiency: the highest mobilization efficiencies were obtained using recipient cells that were in the exponential growth phase. Furthermore, elimination of the origin of transfer from pLS20cat enhanced the mobilization. Conclusions We describe a novel method of plasmid mobilization into G. kaustophilus recipient from B. subtilis donor depending on the helper function of pLS20cat, which enables simple, rapid, and easy transformation of the thermophilic Gram-positive bacterium.
URIhttps://doi.org/10.1186/s12934-018-0969-9
http://hdl.handle.net/10261/168855
Aparece en las colecciones: Colección Biomed Central-Chemistry Central-Springer Open
Ficheros en este ítem:
Fichero Descripción Tamaño Formato  
12934_2018_Article_969.pdf1,22 MBAdobe PDFVista previa
Visualizar/Abrir
Mostrar el registro completo
 


NOTA: Los ítems de Digital.CSIC están protegidos por copyright, con todos los derechos reservados, a menos que se indique lo contrario.