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Published on: February 8, 2016
High-efficiency electrotransformation of the yeast Schwanniomyces occidentalis
P Costaglioli1, E Meilhoc, J M Masson
1Institut National des Sciences Appliquées, CNRS, Toulouse, France.
Abstract:
A method has been developed for introducing heterologous DNA rapidly and efficiently by electropermeabilization into the yeast Schwanniomyces occidentalis. A transformation efficiency as high as 2 x 10(5) transformants/microgram of plasmid DNA was obtained with a square-wave electric pulse of 2.17 kV/cm during 18 ms. Small quantities of DNA (5 ng) can be used to transform 3 x 10(8) cells. The main parameters which have been optimized are: presence of adenine in the culture medium, pretreatment of the cells with dithiothreitol during the exponential growth phase of the cells, amount of cells treated, and pulse-field strength and duration. Competent cells can be stored to allow electrotransformation whenever needed.
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