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.
Reference:
Van Zyl, WH, Lodolo, EJ and Gericke, M. 1993. Conversion of homothallic yeast to heterothallism through to gene disruption. Current Genetics, vol. 23(4), pp 290-294
Van Zyl, W., Lodolo, E., & Gericke, M. (1993). Conversion of homothallic yeast to heterothallism through to gene disruption. http://hdl.handle.net/10204/831
Van Zyl, WH, EJ Lodolo, and M Gericke "Conversion of homothallic yeast to heterothallism through to gene disruption." (1993) http://hdl.handle.net/10204/831
Van Zyl W, Lodolo E, Gericke M. Conversion of homothallic yeast to heterothallism through to gene disruption. 1993; http://hdl.handle.net/10204/831.
