Electroporation at elevated temperatures substantially improves transformation efficiency of slow-growing mycobacteria

• Published in: FEMS microbiology letters Vol. 145; no. 1; pp. 101 - 105
• Main Authors: Wards, Barry J., Collins, Des M.
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 15.11.1996
• Subjects: Electroporation; Mycobacteria; Transformation efficiency; Tuberculosis; Electroporation; Tuberculosis; Mycobacteria; Transformation efficiency
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1016/0378-1097(96)00394-1

The effect of electroporation temperature, biochemical pretreatment of cells and stage of culture on electroporation efficiency for slow-growing mycobacteria were investigated. The efficiency of transformation into Mycobacterium tuberculosis, Mycobacterium bovis and Mycobacterium intracellulare increased markedly with temperature. In contrast, the efficiency of transformation into Mycobacterium smegmatis, a fast-growing species, was higher at 0°C and decreased with temperature. While stage of culture had little effect, a further increase in efficiency of 2–4-fold was obtained following glycine or ethionamide pretreatment. Electroporation at 37°C has been chosen as a standard condition for slow-growing species as it usually resulted in a transformation efficiency several orders of magnitude higher than that obtained at 0°C.