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, 145 (1), 101-5

Electroporation at Elevated Temperatures Substantially Improves Transformation Efficiency of Slow-Growing Mycobacteria

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Electroporation at Elevated Temperatures Substantially Improves Transformation Efficiency of Slow-Growing Mycobacteria

B J Wards et al. FEMS Microbiol Lett.

Abstract

The effects 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 degree 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 degrees 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 degree C.

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