title: The phage integrase vector pIPI03 allows RecA-independent, site-specific labelling of Staphylococcus lugdunensis strains.
abstract: Staphylococcus lugdunensis is a coagulase negative staphylococcus that is a commensal of man and an opportunistic pathogen. A site-specific integrative plasmid for the use in S. lugdunensis was constructed and validated. The integrase gene ccrB of bacteriophage ϕSL01 together with its attachment site was cloned into the thermosensitive plasmid pIMAY. The resulting plasmid pIPI03 integrated RecA-independently, site-specifically and irreversibly into the S. lugdunensis chromosome. Two IPTG-inducible antibiotic resistance determinants were cloned into pIPI03 and the derivatives were used to construct strains suitable for competitive growth experiments in both in vitro and in vivo.
authors.affiliation: Microbiology Department, Moyne Institute of Preventive Medicine, School of Genetics and Microbiology, Trinity College Dublin, Dublin 2, Ireland.
authors.forename: Simon
authors.initials: S
authors.lastname: Heilbronner
authors.forename: Ian R
authors.initials: IR
authors.lastname: Monk
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Foster
mesh_terms: D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D024881:Drug Resistance, Bacterial / Q000235:genetics; D005818:Genetic Engineering; D005822:Genetic Vectors; D019426:Integrases / Q000235:genetics*; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D057787:Staphylococcus lugdunensis / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D014764:Viral Proteins; D007544:Isopropyl Thiogalactoside; D019426:Integrases
keywords: Bacteriophage; Plasmid vector; Staphylococcus lugdunensis; recA mutant