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Meiosis pp 235-252 | Cite as

Using Schizosaccharomyces pombe Meiosis to Analyze DNA Recombination Intermediates

  • Randy W. Hyppa
  • Gerald R. Smith
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 557)

Abstract

The fission yeast Schizosaccharomyces pombe has many biological characteristics that make it an ideal model organism for the study of meiosis. A nearly synchronous meiosis is one of the most important. Under certain environmental and genetic conditions, large cultures of S. pombe can be induced to undergo meiosis in a timely and predictable manner that allows for changes in the DNA to be observed and analyzed by gel electrophoresis. Initiation of meiotic recombination via programmed DNA double-strand breaks, the formation of joint molecule recombination intermediates, and the resolution of these intermediates into crossover DNA products can all be seen with consistent timing during the progression of a synchronous meiotic induction. The timing of recombination events, the genetic requirements for the formation and disappearance of recombination intermediates, and the analysis of the DNA structures of those intermediates allow a comparison of meiotic recombination in fission yeast with that in the only other species similarly studied, the budding yeast Saccharomyces cerevisiae.

Key words

Fission yeast Schizosaccharomyces pombe meiotic induction DNA double-strand breaks joint molecules Holliday junctions intersister interhomolog crossover pulsed-field gel electrophoresis two-dimensional gel electrophoresis 

Notes

Acknowledgments

We are grateful to Sue Amundsen and Gareth Cromie for helpful comments on the manuscript. Our laboratory is supported by research grants GM031693 and GM032194 from the National Institutes of Health.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Randy W. Hyppa
    • 1
  • Gerald R. Smith
    • 1
  1. 1.Division of Basic Sciences, Fred Hutchinson Cancer Research CenterSeattleUSA

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