通过破坏HO基因将同宗配合酵母转化为异宗配合。

Conversion of homothallic yeast to heterothallism through HO gene disruption.

作者信息

van Zyl W H, Lodolo E J, Gericke M

机构信息

Division of Food Science and Technology, CSIR, Pretoria, South Africa.

出版信息

Curr Genet. 1993;23(4):290-4. doi: 10.1007/BF00310889.

DOI:10.1007/BF00310889

PMID:8385580

Abstract

A simple method was developed for the conversion of homothallic Saccharomyces cerevisiae yeast strains to heterothallism through HO gene disruption. An integrative ho::neo disrupted allele was constructed by cloning a dominant selectable marker, the bacterial neo gene, within the HO gene. Transformation of a homothallic diploid S. cerevisiae strain with plasmid DNA containing the ho::neo allele yielded G418-resistant yeast transformants in which one of the HO alleles was replaced by the disrupted ho::neo allele. Meiotic tetrad analysis of four-spored asci from these G418-resistant transformants gave rise to haploid heterothallic and diploid homothallic tetrad progeny. The presence of the ho::neo and HO alleles in the heterothallic and homothallic progeny was confirmed by Southern-blot analysis.

摘要

通过破坏HO基因，开发了一种将同宗配合的酿酒酵母菌株转化为异宗配合的简单方法。通过将显性选择标记细菌新霉素基因克隆到HO基因中，构建了整合型ho::neo破坏等位基因。用含有ho::neo等位基因的质粒DNA转化同宗配合的二倍体酿酒酵母菌株，产生了对G418抗性的酵母转化体，其中一个HO等位基因被破坏的ho::neo等位基因所取代。对这些G418抗性转化体的四孢子子囊进行减数分裂四分体分析，产生了单倍体异宗配合和二倍体同宗配合的四分体后代。通过Southern杂交分析证实了异宗配合和同宗配合后代中ho::neo和HO等位基因的存在。