Targeted gene disruption in Francisella tularensis by group II introns

Stephen A. Rodriguez, Greg Davis, Karl E. Klose

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Francisella tularensis is a highly infectious Gram-negative bacterium that is the causative agent of tularemia. Very little is known about the molecular mechanisms responsible for F. tularensis virulence, in part due to the paucity of genetic tools available for the study of F. tularensis. We have developed a gene knockout system for F. tularensis that utilizes retargeted mobile group II introns, or "targetrons". These targetrons disrupt both single and duplicated target genes at high efficiency in three different F. tularensis subspecies. Here we describe in detail the targetron-based method for insertional mutagenesis of F. tularensis genes, which should facilitate a better understanding of F. tularensis pathogenesis. Group II introns can be adapted to inactivate genes in bacteria for which few genetic tools exist, thus providing a powerful tool to study the genetic basis of bacterial pathogenesis.

Original languageEnglish (US)
Pages (from-to)270-274
Number of pages5
JournalMethods
Volume49
Issue number3
DOIs
StatePublished - Nov 1 2009
Externally publishedYes

Keywords

  • Francisella tularensis
  • Group II intron
  • Mutagenesis
  • Targetron
  • Tularemia

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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