An improved transformation protocol for the human fungal pathogen Candida albicans

被引:176
作者
Walther, A
Wendland, J
机构
[1] Univ Jena, Hans Knoll Inst Naturstoff Forsch, Jr Res Grp Growth Control Fungal Pathogens, D-07745 Jena, Germany
[2] Univ Jena, Dept Microbiol, D-07745 Jena, Germany
关键词
transformation; lithium acetate; integration; heat shock; yeast;
D O I
10.1007/s00294-002-0349-0
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Commonly used protocols for the transformation of the dimorphic human fungal pathogen Candida albicans rely on established methods for the yeast Saccharomyces cerevisiae. With respect to transformation efficiency, however, there is a great difference between these two organisms when using the lithium acetate procedure. Here we present a modified version of this protocol for use with C. albicans. Among the different parameters tested, two turned out to be particularly relevant and, when combined, resulted in an up to 10-fold increase in transformation efficiency (400-500 integrative transformants) compared with previous protocols: first, adjusting the heat shock applied to the cells to 44 degreesC for C. albicans instead of 42 degreesC for S. cerevisiae and, second, treating C. albicans cells with lithium acetate in an overnight incubation instead of for 30 min as used for S. cerevisiae. With these modifications, the lithium acetate procedure becomes a very efficient and reliable tool for C. albicans transformation.
引用
收藏
页码:339 / 343
页数:5
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