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Meiosis pp 55-63 | Cite as

Measurement of Spatial Proximity and Accessibility of Chromosomal Loci in Saccharomyces cerevisiae Using Cre /loxP Site-Specific Recombination

  • Doris Lui
  • Sean M. Burgess
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 557)

Abstract

Several methods have been developed to measure interactions between homologous chromosomes during meiosis in budding yeast. These include cytological analysis of fixed, spread nuclei using fluorescence in situ hybridization (FISH) (1, 2), visualization of GFP-labeled chromosomal loci in living cells (3), and Chromosome-Conformation Capture (3C) (4). Here we describe a quantitative genetic assay that uses exogenous site-specific recombination to monitor the level of homolog associations between two defined loci in living cells of budding yeast (5). We have used the Cre/loxP assay to genetically dissect nuclear architecture and meiotic homolog pairing in budding yeast. Data obtained from this assay report on the relative spatial proximity or accessibility of two chromosomal loci located within the same strain and can be compared to measurements from different mutated strains.

Key words

budding yeast meiosis homolog pairing chromosome collision assay site-specific recombination Cre/loxP 

Notes

Acknowledgments

The authors would like to give a special thanks to Tamara Peoples-Holst, Eric Dean, and Joshua Chang Mell for their contribution to the optimization of this protocol over the years. This work has been supported by the National Institutes of Health (NIH) grant NIH R01 GM075119 (S.M.B), the American Cancer Society RSG-01-053-01-CCG (S.M.B), and the NIH-Environmental Health Sciences training grant NIH T 32 ES07059 (D.Y.L).

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Copyright information

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

Authors and Affiliations

  • Doris Lui
    • 1
  • Sean M. Burgess
    • 1
  1. 1.Section of Molecular and Cellular BiologyCollege of Biological Sciences, University of CaliforniaDavisUSA

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