Use of bacteriophage lambda recombination functions to promote gene replacement in Escherichia coli

J Bacteriol. 1998 Apr;180(8):2063-71. doi: 10.1128/JB.180.8.2063-2071.1998.

Abstract

Replacement of Escherichia coli's RecBCD function with phage lambda's Red function generates a strain whose chromosome recombines with short linear DNA fragments at a greatly elevated rate. The rate is at least 70-fold higher than that exhibited by a recBC sbcBC or recD strain. The value of the system is highlighted by gene replacement with a PCR-generated DNA fragment. The deltarecBCD::Plac-red kan replacement allele can be P1 transduced to other E. coli strains, making the hyper-Rec phenotype easily transferable.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacteriophage lambda / genetics*
  • Calcium / pharmacology
  • Chromosomes, Bacterial / genetics*
  • Cloning, Molecular / methods
  • DNA Primers
  • DNA, Superhelical
  • Escherichia coli / drug effects
  • Escherichia coli / genetics*
  • Escherichia coli / virology
  • Gene Targeting / methods*
  • Genotype
  • Operon
  • Plasmids
  • Polymerase Chain Reaction
  • Recombinant Fusion Proteins / biosynthesis
  • Recombination, Genetic*
  • Restriction Mapping

Substances

  • DNA Primers
  • DNA, Superhelical
  • Recombinant Fusion Proteins
  • Calcium