酿酒酵母 2μmDNA 的转化修复。
Curing of Saccharomyces cerevisiae 2-μm DNA by transformation.
机构信息
Institut für Mikrobiologie, Universität Düsseldorf, Universitätsstraße 1, D-4000, Düsseldorf, Germany.
出版信息
Curr Genet. 1981 May;3(2):83-9. doi: 10.1007/BF00365710.
A general procedure for the curing of 2-μm in Saccharomyces cerevisiae is described. The method is based on the displacement of endogenous 2-μm DNA by the recombinant plasmid pMP78-1, which carries the yeast leu2 gene and the 2 -μm DNA replicon, but cannot be maintained stably in a yeast cell without endogenous 2-μm DNA. After transformation with pMP78-1 cells are grown selectively to displace 2-μm DNA. During the non-selective growth which follows, plasmid pMP78-1 is lost and up to 100% of the cells completely lack plasmids. In conjunction with a kanamycin resistance marker, as present in plasmid pMP81, this method should be applicable to cure any wild-type yeast strain. The stability of recombinant plasmids in cir (+) and cir (0) strains has been compared.
描述了一种在酿酒酵母中修复 2μm 缺陷的通用程序。该方法基于通过携带酵母亮氨酸基因和 2μmDNA 复制子的重组质粒 pMP78-1 替换内源性 2μmDNA，但如果没有内源性 2μmDNA，该质粒在酵母细胞中不能稳定维持。在用 pMP78-1 转化后，细胞进行选择性生长以替换 2μmDNA。在随后的非选择性生长中，质粒 pMP78-1 丢失，高达 100%的细胞完全缺乏质粒。与存在于质粒 pMP81 中的卡那霉素抗性标记结合使用，该方法应该适用于修复任何野生型酵母菌株。已经比较了重组质粒在 cir (+) 和 cir (0) 菌株中的稳定性。
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