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DNA Recombination

Volume 745 of the series Methods in Molecular Biology pp 151-172

Date:

Genetic and Molecular Analysis of Mitotic Recombination in Saccharomyces cerevisiae

  • Belén Gómez-GonzálezAffiliated withCentro Andaluz de Biología Molecular y Medicina Regenerativa, Universidad de Sevilla-CSIC Email author 
  • , José F. RuizAffiliated withCentro Andaluz de Biología Molecular y Medicina Regenerativa, Universidad de Sevilla-CSIC
  • , Andrés AguileraAffiliated withCentro Andaluz de Biología Molecular y Medicina Regenerativa, Universidad de Sevilla-CSIC

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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