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Reporter Gene-Based Recombination Lines for Studies of Genome Stability

  • Palak Kathiria
  • Igor Kovalchuk
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 631)

Abstract

Homologous recombination is a double-strand break repair mechanism operating in somatic cells and involved in meiotic crossovers in plants. It is responsible for the maintenance of genome stability and thus plays a crucial role in adaptation to stress. Recombination between homologous loci is believed to be regulated in part by epigenetic machinery such as methylation. Therefore, the recombination frequency at a specific locus can reflect the chromatin status.

Several reporter gene-based recombination constructs have been developed to study HR frequencies in plants. Among them, the luciferase and beta-glucuronidase-based recombination reporter systems are the most widely used. Here, we explain how reporter gene recombination assays operate and in which applications they are used. We also present a conceptually new system for analysis of sequence-specific recombination frequency. These assays can be effectively used for analysis of locus-specific endogenous and stress-induced recombination frequencies.

Key words

Homologous recombination Reporter gene recombination constructs Luciferase Beta-glucuronidase Intramolecular recombination Intermolecular recombination 

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Palak Kathiria
    • 1
  • Igor Kovalchuk
    • 1
  1. 1.Department of Biological SciencesUniversity of LethbridgeLethbridgeCanada

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