A simple method for isolating disomic strains of Saccharomyces cerevisiae

Yeast. 2008 May;25(5):321-6. doi: 10.1002/yea.1590.

Abstract

A simple method to select disomic (N + 1) strains that should be applicable for almost any chromosome in Saccharomyces cerevisiae is presented. A diploid heterozygous for a KanMX knock-out mutation in an essential gene is sporulated and viable geneticin (G418)-resistant colonies selected. Disomic products of a missegregation or non-disjunction event containing a copy of both the wild-type essential gene and its complementary KanMX knock-out allele make up most of the viable colonies. This method has been used to isolate disomic haploids for a variety of chromosomes. It is appropriately named MARV (for missegregation-associated restoration of viability) and is easily adaptable to virtually any strain.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Chromosomes, Fungal / genetics*
  • Diploidy
  • Gentamicins / pharmacology
  • Haploidy
  • Mutagenesis, Insertional / methods*
  • Open Reading Frames
  • Polymerase Chain Reaction
  • Recombination, Genetic
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / physiology
  • Saccharomyces cerevisiae Proteins / genetics
  • Spores, Fungal / growth & development
  • Transformation, Genetic

Substances

  • Gentamicins
  • Saccharomyces cerevisiae Proteins
  • antibiotic G 418