Molecular and General Genetics MGG

, Volume 223, Issue 2, pp 241–248

Transformation and recombination in rad mutants of Saccharomyces cerevisiae

  • John R. Simon
  • Peter D. Moore
Article

DOI: 10.1007/BF00265060

Cite this article as:
Simon, J.R. & Moore, P.D. Molec. Gen. Genet. (1990) 223: 241. doi:10.1007/BF00265060

Summary

Disruption/deletion mutations in genes of the RAD52 epistasis group of Saccharomyces cerevisiae were examined for their effects on recombination between single-and double-stranded circular DNA substrates and chromosomal genes in a transformation assay. In rad50 mutants there was a small reduction in recombination with single-stranded DNA at the leu2-3, 112 allele; in addition there was an almost complete elimination of recombination at trpl-1 for both single- and double-stranded DNA. Reintroduction of a wild-type RAD50 gene on a replicating plasmid carrying CEN4 restored recombinational competence at trpl-1, indicating that rad50 is defective in gene replacement of this allele. In rad52 mutants a reduction of 30%-50% in recombination involving either single- or double-stranded circular DNA was observed in each experiment when compared to the wild type. This reduction of recombination in rad52 mutants was similar for recombination at the ura352 mutant locus where only integration events have been observed, and at the trpl-1 mutant locus, where recombination occurs predominantly by gene replacement. Neither the rad54 nor the rad57 mutations had a significant effect on recombination with single- or double-stranded DNA substrates.

Key words

Homologous RecombinationTransformationSaccharomyces CerevisiaePlasmidsrad mutants

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • John R. Simon
    • 1
  • Peter D. Moore
    • 1
  1. 1.Department of Microbiology and ImmunologyUniversity of Illinois College of MedicineChicagoUSA
  2. 2.Department of Biological ChemistryUCLA School of MedicineLos AngelesUSA
  3. 3.Department of GeneticsUniversity of Illinois College of MedicineChicagoUSA