Gene replacement in Staphylococcus carnosus and Staphylococcus xylosus.
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.
Brückner
R
R
Mikrobielle Genetik, Universität Tübingen, Germany. reinhold.brueckner@uni-tuebingen.de
D000483:Alleles; D004352:Drug Resistance, Microbial / Q000235:genetics; D004917:Erythromycin / Q000494:pharmacology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006026:Glycoside Hydrolases / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics; D017852:Phosphotransferases (Nitrogenous Group Acceptor) / Q000235:genetics; D013210:Staphylococcus / Q000235:genetics*; D043324:beta-Fructofuranosidase
D004917:Erythromycin; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; D017852:Phosphotransferases (Nitrogenous Group Acceptor); C022360:phosphoenolpyruvate-protein phosphotransferase; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase