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一种用于制备通过电穿孔进行转化的酵母细胞的改进方案。

An improved protocol for the preparation of yeast cells for transformation by electroporation.

作者信息

Thompson J R, Register E, Curotto J, Kurtz M, Kelly R

机构信息

Department of Biochemistry, Merck & Co., Rahway, NJ 07065, USA.

出版信息

Yeast. 1998 Apr 30;14(6):565-71. doi: 10.1002/(SICI)1097-0061(19980430)14:6<565::AID-YEA251>3.0.CO;2-B.

Abstract

Pretreatment of yeast cells with lithium acetate (LiAc) and dithiothreitol (DTT) enhances the frequency of transformation by electroporation. The method shows improvements of 6-67-fold in wild-type strains derived from commonly used Saccharomyces cerevisiae genetic backgrounds. In addition, 15-300-fold improvement in transformation frequency was achieved with several mutant strains of S. cerevisiae that transformed poorly by conventional procedures. Both DTT and lithium acetate were necessary for maximal transformation frequencies. Pretreatment with lithium and DTT also resulted in an approximately 3-5-fold increase in the electroporation transformation frequency of the pathogenic fungus Candida albicans.

摘要

用醋酸锂(LiAc)和二硫苏糖醇(DTT)对酵母细胞进行预处理可提高电穿孔转化的频率。该方法在源自常用酿酒酵母遗传背景的野生型菌株中显示出6至67倍的改进。此外,对于几种通过传统方法转化效率较低的酿酒酵母突变菌株,转化频率提高了15至300倍。DTT和醋酸锂对于实现最大转化频率都是必需的。用锂和DTT进行预处理还使致病真菌白色念珠菌的电穿孔转化频率提高了约3至5倍。

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