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

Simon Heilbronner1, Ian R Monk, Timothy J Foster

  • 1Microbiology Department, Moyne Institute of Preventive Medicine, School of Genetics and Microbiology, Trinity College Dublin, Dublin 2, Ireland.

Plasmid
|September 18, 2013
PubMed

Insights

Researchers developed a novel integrative plasmid for Staphylococcus lugdunensis, enabling precise genetic modification. This tool facilitates the study of this opportunistic pathogen

Area of Science:

  • Microbiology
  • Bacteriology
  • Molecular Biology

Background:

  • Staphylococcus lugdunensis is a common human commensal and opportunistic pathogen.
  • Genetic manipulation of S. lugdunensis is crucial for understanding its pathogenic mechanisms.

Purpose of the Study:

  • To construct and validate a site-specific integrative plasmid for S. lugdunensis.
  • To develop tools for studying S. lugdunensis in various experimental settings.

Main Methods:

  • Cloning of bacteriophage ϕSL01 integrase gene (ccrB) and attachment site into a thermosensitive plasmid (pIMAY).
  • Construction of the integrative plasmid pIPI03, demonstrating RecA-independent, site-specific chromosomal integration.
  • Introduction of IPTG-inducible antibiotic resistance determinants into pIPI03.

Main Results:

  • The plasmid pIPI03 achieved irreversible, site-specific integration into the S. lugdunensis chromosome.
  • Engineered strains facilitated competitive growth experiments.
  • The developed system allows for precise genetic manipulation and strain construction.

Conclusions:

  • A functional site-specific integrative plasmid system for Staphylococcus lugdunensis has been established.
  • This novel tool enhances the capability for genetic studies of S. lugdunensis.
  • The system supports in vitro and in vivo competitive growth experiments.
Keywords:
BacteriophagePlasmid vectorStaphylococcus lugdunensisrecA mutant

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