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KMID : 0364820130490010001
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Korean Journal of Microbiology
2013 Volume.49 No. 1 p.1 ~ p.7
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Identification of Meiotic Recombination Intermediates in Saccharomyces cerevisiae
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Sung Young-Jin
Yoon Sang-Wook
Kim Keun-Pil
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Abstract
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During meiosis, genetic recombinants are formed by homologous recombination accompanying with the
programmed double-strand breaks (DSBs) and strand exchanges between homologous chromosomes. The
mechanism is generated by recombination intermediates such as single-end invasions (SEIs) and double-Holliday
junctions (dHJs), and followed by crossover (CO) or non-crossover (NCO) products. Our study was focused on the
analysis of meiotic recombination intermediates (DSBs, SEIs, and dHJs) and final recombination products (CO and
NCO). We identified these meiotic recombination intermediates using DNA physical analysis under HIS4LEU2 “hot
spot” system in budding yeast, Saccharomyces cerevisiae. For DNA physical analysis, when the hot spot locus is
recognized by restriction enzyme from synchronous meiotic cells, the fragmented DNA that are forming
recombination intermediates can be detected and quantified through Southern hybridization analysis. Our study
suggests that this system can analyze the structural change of recombination intermediates during DSB-SEI
transition, double-Holiday junctions and crossover/non-crossover products in meiosis
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KEYWORD
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Saccharomyces cerevisiae, crossover/non-crossover, double-strand breaks, meiotic recombination
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