Development of novel plasmid vectors and a promoter trap system in Francisella tularensis compatible with the pFLN10 based plasmids

David A. Rasko, Carlos D. Esteban, Vanessa Sperandio

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Francisella tularensis is a category A bioterror pathogen which in some cases can cause a severe and fatal human infection. Very few virulence factors are known in this species due to the difficulty in working with it as well as the lack of tools for genetic manipulation. This work describes the construction of a shuttle vector that can replicate in Escherichia coli and F. tularensis as well as two distinct promoter trap constructs based on the shuttle vector backbone. Replication in F. tularensis is based on the promiscuous origin of replication from the Staphylococcus aureus plasmid pC194. We demonstrate the novel plasmids can coexist with established F. tularensis vectors based on the pFNL10 plasmid, the current workhorse of F. tularensis genetics. Our promoter trap can identify promoters that are activated during intracellular growth and survival. These new vectors provide additional tools for the genetic manipulation of F. tularensis.

Original languageEnglish (US)
Pages (from-to)159-166
Number of pages8
JournalPlasmid
Volume58
Issue number2
DOIs
StatePublished - Sep 1 2007

Keywords

  • Francisella tularensis
  • Promoter trap
  • Shuttle vector

ASJC Scopus subject areas

  • Molecular Biology

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