Eukaryotic DNA mismatch repair in vitro

Fenghua Yuan, Limin Song, Fengsong Liu, Liya Gu, Yanbin Zhang

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Mismatch repair corrects biosynthetic errors generated during DNA replication. Mismatch repair deficiency causes a mutator phenotype and directly underlies hereditary nonpolyposis colorectal cancer and some sporadic cancers. Because of remarkably high conservation of the mismatch repair machinery between the budding yeast (Saccharomyces cerevisiae) and humans, the study of mismatch repair in yeast has provided tremendous insights into the mechanisms of this repair pathway in humans. Here we describe a set of practical protocols for how to prepare the yeast and HeLa cell-free nuclear extracts and site-specific DNA mismatch substrates, and how to carry out the in vitro mismatch repair assay. We validated the yeast cell-free system by the mismatch repair deficient strain (Δmsh2) and the complementation assay with purified yeast MutSα.

Original languageEnglish (US)
Title of host publicationDNA Repair Protocols
EditorsLotte Bjergbaek
Pages149-162
Number of pages14
DOIs
StatePublished - Nov 12 2012

Publication series

NameMethods in Molecular Biology
Volume920
ISSN (Print)1064-3745

Keywords

  • Cancer
  • Cell-free nuclear extract
  • Human
  • In vitro DNA mismatch repair
  • MMR
  • Saccharomyces cerevisiae
  • Yeast

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Yuan, F., Song, L., Liu, F., Gu, L., & Zhang, Y. (2012). Eukaryotic DNA mismatch repair in vitro. In L. Bjergbaek (Ed.), DNA Repair Protocols (pp. 149-162). (Methods in Molecular Biology; Vol. 920). https://doi.org/10.1007/978-1-61779-998-3-11