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Visualization of Human Dmc1 Presynaptic Filaments

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DNA Recombination

Part of the book series: Methods in Molecular Biology ((MIMB,volume 745))

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Abstract

Meiosis is initiated by the programmed formation of DNA double-strand breaks (DSBs). These DSBs are repaired by homologous recombination to promote crossover formation that ensures proper chromosomal segregation in meiosis. hRad51 and hDmc1 are two human recombinases present during meiosis that are homologous to the RecA recombinase from Escherichia coli. The hRad51 and hDmc1 recombinases bind the nucleolytically processed ends of the DSB forming a presynaptic filament. Formation of the presynaptic filament is necessary for the search for homology and the progression of recombination. In this chapter, we provide a method to purify hDmc1 and prepare samples for visualizing hDmc1 nucleoprotein presynaptic filaments via transmission electron microscopy.

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Abbreviations

HR:

homologous recombination

ss:

single stranded

DSB:

DNA double-strand break

NTA:

nitro triacetic acid

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Acknowledgments

This work was supported by National Science Foundation/EPSCoR grant 2004 RII-EPS-0447660 and Clemson University.

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Authors and Affiliations

  1. Department of Genetics and Biochemistry, Clemson University, Clemson, SC, 29634, USA

    Michael G. Sehorn & Hilarie A. Sehorn

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  1. Michael G. Sehorn
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  2. Hilarie A. Sehorn
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Corresponding author

Correspondence to Michael G. Sehorn .

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Editors and Affiliations

  1. MRC Genome Damage and Stability Centre, University of Sussex, Science Park Road, Brighton, BN1 9RQ, East Sussex, United Kingdom

    Hideo Tsubouchi

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Sehorn, M.G., Sehorn, H.A. (2011). Visualization of Human Dmc1 Presynaptic Filaments. In: Tsubouchi, H. (eds) DNA Recombination. Methods in Molecular Biology, vol 745. Humana Press. https://doi.org/10.1007/978-1-61779-129-1_28

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  • DOI: https://doi.org/10.1007/978-1-61779-129-1_28

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-128-4

  • Online ISBN: 978-1-61779-129-1

  • eBook Packages: Springer Protocols

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