Conversion of homothallic yeast to heterothallism through HO gene disruption |
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| Authors: | W H van Zyl E J Lodolo M Gericke |
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| Institution: | (1) Division of Food Science and Technology, CSIR, P.O. Box 395, 0001 Pretoria, South Africa;(2) Present address: Microbiology Department, Stellenbosch University, 7600 Stellenbosch, South Africa;(3) Present address: Research and Development, The South African Breweries, P.O. Box 782178, 2146 Sandton, South Africa |
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| 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 Souther-blot analysis. |
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| Keywords: | S cerevisiae Homothallism/heterothallism HO gene disruption G418 resistance |
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