Reconstituting the 4-Strand DNA Strand Exchange

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Resumen

Proteins of the Rad51 family play a key role in homologous recombination by carrying out DNA strand exchange. Here, we present the methodology and the protocols for the 4-strand exchange between gapped circular DNA and homologous linear duplex DNA promoted by human Rad51 and Escherichia coli RecA orthologs. This reaction includes formation of joint molecules and their extension by branch migration in a polar manner. The presented methodology may be used for reconstitution of the medial-to-late stages of homologous recombination in vitro as well as for investigation of the mechanisms of branch migration by helicase-like proteins, e.g., Rad54, BLM, or RecQ1.

Idioma originalEnglish (US)
Título de la publicación alojadaMethods in Enzymology
EditorialAcademic Press Inc.
Páginas285-305
Número de páginas21
DOI
EstadoPublished - 2018
Publicado de forma externa

Serie de la publicación

NombreMethods in Enzymology
Volumen600
ISSN (versión impresa)0076-6879
ISSN (versión digital)1557-7988

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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