Enhancement of targeted homologous recombination in Mycoplasma mycoides subsp. capri by inclusion of heterologous recA

Appl Environ Microbiol. 2010 Oct;76(20):6951-4. doi: 10.1128/AEM.00056-10. Epub 2010 Aug 27.

Abstract

A suicide plasmid, pExp1-ctpA::tetM-recAec, employing recA from Escherichia coli and tetM as a selection marker, was used to generate ctpA knockout mutants in Mycoplasma mycoides subsp. capri through targeted gene disruption. Inclusion of E. coli recA greatly enhanced both the consistency and the recovery of mutants generated by homologous recombination.

Publication types

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

MeSH terms

  • Algal Proteins
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Carboxypeptidases / genetics
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Gene Knockout Techniques
  • Gene Targeting / methods*
  • Genetic Vectors
  • Mycoplasma mycoides / genetics*
  • Plasmids
  • Proprotein Convertases / genetics
  • Rec A Recombinases / genetics
  • Rec A Recombinases / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Recombination, Genetic*
  • Selection, Genetic
  • Tetracycline Resistance

Substances

  • Algal Proteins
  • Bacterial Proteins
  • Recombinant Proteins
  • Tet M resistance protein, Bacteria
  • Rec A Recombinases
  • Carboxypeptidases
  • Proprotein Convertases
  • photosystem II, D1 polypeptide carboxy-terminal processing protease