f Transformation of mycobacterial species using hygromycin resistance as selectable marker
- Authors: Thomas R. Garbe†, Jaya Barathi, Simona Barnini, Ying Zhang, Christiane Abou-Zeid, Dan Tang, Rama Mukherjee, Douglas B. Young‡
- Author for correspondence: Douglas B. Young. Tel: +44 71 723 1252. Fax: +44 71 262 6299.
- Microbiology, January 1994 140: 133-138, doi: 10.1099/13500872-140-1-133
- Subject: Genetics And Molecular Biology
- Published Online:
Summary: Electroporation with shuttle plasmids carrying a kanamycin resistance gene as a selectable marker failed to generate transformants in two mycobacterial species currently being used in human vaccine trials (Mycobacterium w and Mycobacterium vaccae). In contrast, efficient transformation [103-105 transformants (μg DNA)−1] was obtained using novel vectors with selection based on expression of resistance to hygromycin. The hygromycin resistance vector was also found to be more efficient than kanamycin resistance vectors for transformation of Mycobacterium smegmatis and Mycobacterium bovis BCG. The hygromycin resistance vector was used to overexpress superoxide dismutase of Mycobacterium tuberculosis in M. vaccae in a form suitable for detailed structural analysis. The potential use of this approach for generation of novel recombinant mycobacterial vaccines is discussed.
Present address: Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati Medical Center, Cincinnati, Ohio, USA.
Present address: Department of Medical Microbiology, St '1s Hospital Medical School, Norfolk Place, London W2 1PG, UK.
© Society for General Microbiology 1994 | Published by the Microbiology Society
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