Gene replacement in Staphylococcus carnosus and Staphylococcus xylosus

FEMS Microbiol Lett. 1997 Jun 1;151(1):1-8. doi: 10.1111/j.1574-6968.1997.tb10387.x.

Abstract

A system for high-efficiency gene replacement in Staphylococcus carnosus and Staphylococcus xylosus has been developed, that is based on temperature-sensitive Escherichia coli-Staphylococcus shuttle vectors for fragment delivery and erythromycin resistance cassettes to facilitate selection of genomic copies of disrupted genes. The approach was tested by constructing a phosphotransferase-deficient mutant of S. carnosus and an S. xylosus mutant strain unable to utilize sucrose. Allelic replacements were observed at rather high frequencies, ranging from approximately 10% for the ptsI gene in S. carnosus up to 50% for the scrB gene in S. xylosus. These differences most likely reflect the length of homology rather than strain-specific variations in recombination efficiencies. Apart from the staphylococcal species tested in this study, the system appears to be applicable in other staphylococci.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Drug Resistance, Microbial / genetics
  • Erythromycin / pharmacology
  • Gene Transfer Techniques*
  • Genetic Vectors*
  • Glycoside Hydrolases / genetics
  • Mutagenesis, Site-Directed*
  • Phosphoenolpyruvate Sugar Phosphotransferase System / genetics
  • Phosphotransferases (Nitrogenous Group Acceptor) / genetics
  • Staphylococcus / genetics*
  • beta-Fructofuranosidase

Substances

  • Erythromycin
  • Phosphoenolpyruvate Sugar Phosphotransferase System
  • Phosphotransferases (Nitrogenous Group Acceptor)
  • phosphoenolpyruvate-protein phosphotransferase
  • Glycoside Hydrolases
  • beta-Fructofuranosidase