Reconstituted System for the Examination of Repair DNA Synthesis in Homologous Recombination

Youngho Kwon, James M. Daley, Patrick Sung

Producción científica: Chapter

8 Citas (Scopus)

Resumen

In homologous recombination (HR), DNA polymerase δ-mediated DNA synthesis occurs within the displacement loop (D-loop) that is made by the recombinase Rad51 in conjunction with accessory factors. We describe in this chapter the reconstitution of the D-loop and repair DNA synthesis reactions using purified Saccharomyces cerevisiae HR (Rad51, RPA, and Rad54) and DNA replication (PCNA, RFC, and DNA polymerase δ) proteins and document the role of the Pif1 helicase in DNA synthesis via a migrating DNA bubble intermediate. These reconstituted systems are particularly valuable for understanding the conserved mechanism of repair DNA synthesis dependent on DNA polymerase δ and its cognate helicase in eukaryotic organisms.

Idioma originalEnglish (US)
Título de la publicación alojadaMethods in Enzymology
EditorialAcademic Press Inc.
Páginas307-325
Número de páginas19
DOI
EstadoPublished - 2017
Publicado de forma externa

Serie de la publicación

NombreMethods in Enzymology
Volumen591
ISSN (versión impresa)0076-6879
ISSN (versión digital)1557-7988

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

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