The phage integrase vector pIPI03 allows RecA-independent, site-specific labelling of Staphylococcus lugdunensis strains

Plasmid. 2013 Nov;70(3):377-84. doi: 10.1016/j.plasmid.2013.09.001. Epub 2013 Sep 13.

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.

Keywords: Bacteriophage; Plasmid vector; Staphylococcus lugdunensis; recA mutant.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophages / genetics*
  • Base Sequence
  • Chromosomes, Bacterial*
  • Drug Resistance, Bacterial / genetics
  • Genetic Engineering
  • Genetic Vectors
  • Integrases / genetics*
  • Isopropyl Thiogalactoside / pharmacology
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Plasmids*
  • Staphylococcus lugdunensis / genetics*
  • Viral Proteins / genetics*

Substances

  • Viral Proteins
  • Isopropyl Thiogalactoside
  • Integrases

Associated data

  • GENBANK/KF573190