Mutation detection with MutH, MutL, and MutS mismatch repair proteins.

Published

Journal Article

Escherichia coli methyl-directed mismatch repair is initiated by MutS-, MutL-, and ATP-dependent activation of MutH endonuclease, which cleaves at d(GATC) sites in the vicinity of a mismatch. This reaction provides an efficient method for detection of mismatches in heteroduplexes produced by hybridization of genetically distinct sequences after PCR amplification. Multiple examples of transition and transversion mutations, as well as one, two, and three nucleotide insertion/deletion mutants, have been detected in PCR heteroduplexes ranging in size from 400 bp to 2.5 kb. Background cleavage of homoduplexes is largely due to polymerase errors that occur during amplification, and the MutHLS reaction provides an estimate of the incidence of mutant sequences that arise during PCR.

Full Text

Duke Authors

Cited Authors

  • Smith, J; Modrich, P

Published Date

  • April 30, 1996

Published In

Volume / Issue

  • 93 / 9

Start / End Page

  • 4374 - 4379

PubMed ID

  • 8633074

Pubmed Central ID

  • 8633074

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.93.9.4374

Language

  • eng

Conference Location

  • United States