High efficiency electrotransformation of the yeast Schwanniomyces occidentalis
1994
Costaglioli, P. | Meilhoc, E. | Masson, J.M. (Centre National de la Recherche Scientifique, Toulouse (France). Inst. National des Sciences Appliquees. Lab. de Pharmacologie et Toxicologie Fondamentales)
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 200000 transformants/micro g of plasmid DNA was obtained with a squarewave electric pulse of 2.17 kV/cm during 18 ms. Small quantities of DNA (5 ng) can be used to transform 30000000 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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