Costaglioli P, Meilhoc E, Masson J M
Institut National des Sciences Appliquées, CNRS, Toulouse, France.
Curr Genet. 1994 Dec;27(1):26-30. doi: 10.1007/BF00326575.
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.
已开发出一种通过电穿孔将异源DNA快速高效地导入西方许旺酵母的方法。在18毫秒内施加2.17 kV/cm的方波电脉冲，可获得高达2×10⁵转化子/微克质粒DNA的转化效率。少量DNA（5纳克）可用于转化3×10⁸个细胞。已优化的主要参数包括：培养基中腺嘌呤的存在、在细胞指数生长期用二硫苏糖醇对细胞进行预处理、处理的细胞量以及脉冲场强度和持续时间。感受态细胞可储存起来，以便在需要时进行电转化。
