Genetic and Molecular Analysis of Mitotic Recombination in Saccharomyces cerevisiae

  • Belén Gómez-González
  • José F. Ruiz
  • Andrés Aguilera
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 745)

Abstract

Many systems have been developed for the study of mitotic homologous recombination (HR) in the yeast Saccharomyces cerevisiae at both genetic and molecular levels. Such systems are of great use for the analysis of different features of HR as well as of the effect of mutations, transcription, etc., on HR. Here we describe a selection of plasmid- and chromosome-borne DNA repeat assays, as well as plasmid–chromosome recombination systems, which are useful for the analysis of spontaneous and DSB-induced recombination. They can easily be used in diploid and, most importantly, in haploid yeast cells, which is a great advantage to analyze the effect of recessive mutations on HR. Such systems were designed for the analysis of a number of different HR features, which include the frequency and length of the gene conversion events, the frequency of reciprocal exchanges, the proportion of gene conversion versus reciprocal exchange, or the molecular analysis of sister chromatid exchange.

Key words

Homologous recombination direct repeats inverted repeats sister chromatid exchange gene conversion reciprocal exchange yeast 

Notes

Acknowledgments

We would like to thank H. Gaillard and M. Moriel-Carretero for reading the manuscript and D. Haun for style supervision. We apologize for not citing our colleague’s work due to space limitation. Research in the A. A. laboratory was funded by research grants from the Spanish Ministry of Science and Innovation and the Junta de Andalucía.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Belén Gómez-González
    • 1
  • José F. Ruiz
    • 1
  • Andrés Aguilera
    • 1
  1. 1.Centro Andaluz de Biología Molecular y Medicina RegenerativaUniversidad de Sevilla-CSICSevillaSpain

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