Conversion of homothallic yeast to heterothallism through HO gene disruption.
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.
Division of Food Science and Technology, CSIR, Pretoria, South Africa.
W H
WH
van Zyl
E J
EJ
Lodolo
M
M
Gericke
D000483:Alleles; D015825:Chromosomes, Fungal; D004720:Endonucleases / Q000235:genetics*; D005785:Gene Conversion; D005821:Genetic Techniques; D019868:Kanamycin Kinase; D010770:Phosphotransferases / Q000235:genetics; D010957:Plasmids; D011003:Ploidies; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D014170:Transformation, Genetic
D010770:Phosphotransferases; D019868:Kanamycin Kinase; D004720:Endonucleases