SOE-LRed: A simple and time-efficient method to localize genes with point mutations onto the Escherichia coli chromosome

J Microbiol Methods. 2011 Mar;84(3):479-81. doi: 10.1016/j.mimet.2010.12.020. Epub 2010 Dec 24.

Abstract

We use a powerful method to replace wild-type genes on the chromosome of Escherichia coli. Using a unique form of PCR, we generate easily constructible gene fusions bearing single point mutations. Used in conjunction with homologous recombination, this method eliminates cloning procedures previously used for this purpose.

MeSH terms

  • Artificial Gene Fusion
  • Chromosome Mapping / methods*
  • Chromosomes, Bacterial
  • Escherichia coli / genetics*
  • Escherichia coli Proteins / genetics*
  • Genes, Bacterial*
  • Genetics, Microbial / methods*
  • Molecular Biology / methods
  • Mutant Proteins / genetics*
  • Point Mutation*
  • Polymerase Chain Reaction / methods
  • Recombination, Genetic

Substances

  • Escherichia coli Proteins
  • Mutant Proteins