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Fig. 1 | BMC Microbiology

Fig. 1

From: Transmating: conjugative transfer of a new broad host range expression vector to various Bacillus species using a single protocol

Fig. 1

Steps for transfer of pBACOV-sfGFP from E. coli to Bacillus species by transmating. Triparental conjugation (transmating) requires an acceptor strain (strain of a Bacillus species of choice), a donor strain (e. g. E. coli TOP10 carrying pBACOV-sfGFP) and a helper strain (E. coli HB101 pRK2013). After conjugation on agar plates without selection, the cells are transferred to appropriate antibiotic-containing plates (Pol concentration < MIC of acceptor, Kan concentration > MIC of acceptor) for selection of transmates (a). Plasmid content and the identity of the host strain were confirmed by colony PCR and 16S rRNA gene sequencing (b) before further characterization of the transmates (c). MIC: minimal inhibitory concentration, Kan: kanamycin, Pol: polymyxin B

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BMC Microbiology

ISSN: 1471-2180

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