ArticleApplied and environmental microbiology2012
TetR-based gene regulation systems for Francisella tularensis.
Article in Applied and environmental microbiology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 27 citations in OpenAlex.
- Development of inducible promoter and CRISPRi plasmids functional inJournal of bacteriology · 2024Article
- Mutation ofMicrobiology (Reading, England) · 2024Article
- Tularemia treatment: experimental and clinical data.Frontiers in microbiology · 2023Review
- Francisella tularensis Exploits AMPK Activation to Harvest Host-Derived Nutrients Liberated from Host Lipolysis.Infection and immunity · 2022Article
- Status quo of tet regulation in bacteria.Microbial biotechnology · 2022Review
- Genetic Determinants of Antibiotic Resistance inFrontiers in microbiology · 2021Review
- Article
- Global Analysis of Genes Essential for Francisella tularensis Schu S4 GrowthJournal of bacteriology · 2019Article
- Multiple domains of bacterial and human Lon proteases define substrate selectivity.Emerging microbes & infections · 2018Article
- Zinc Acquisition Mechanisms Differ between Environmental and Virulent Francisella Species.Journal of bacteriology · 2018Article
- Mobilizable Plasmids for Tunable Gene Expression inFrontiers in cellular and infection microbiology · 2018Article
- A Bioluminescent Francisella tularensis SCHU S4 Strain Enables Noninvasive Tracking of Bacterial Dissemination and the Evaluation of Antibiotics in an Inhalational Mouse Model of Tularemia.Antimicrobial agents and chemotherapy · 2016Article
- The Protease Locus of Francisella tularensis LVS Is Required for Stress Tolerance and Infection in the Mammalian Host.Infection and immunity · 2016Article
- Expansion of the tetracycline-dependent regulation toolbox for Helicobacter pylori.Applied and environmental microbiology · 2015Article
- Identifying Francisella tularensis genes required for growth in host cells.Infection and immunity · 2015Article
- Genetic engineering of Francisella tularensis LVS for use as a novel live vaccine platform against Pseudomonas aeruginosa infections.Bioengineered · 2015Article
- Extragenic suppressor mutations in ΔripA disrupt stability and function of LpxA.BMC microbiology · 2014Article
- The contribution of the glycine cleavage system to the pathogenesis of Francisella tularensis.Microbes and infection · 2014Article
- Synthetic promoters functional in Francisella novicida and Escherichia coli.Applied and environmental microbiology · 2014Article
- Francisella tularensis harvests nutrients derived via ATG5-independent autophagy to support intracellular growth.PLoS pathogens · 2013Article
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
There are a number of genetic tools available for studying Francisella tularensis, the etiological agent of tularemia; however, there is no effective inducible or repressible gene expression system. Here, we describe inducible and repressible gene expression systems for F. tularensis based on the Tet repressor, TetR. For the inducible system, a tet operator sequence was cloned into a modified F. tularensis groESL promoter sequence and carried in a plasmid that constitutively expressed TetR. To monitor regulation the luminescence operon, luxCDABE, was cloned under the hybrid Francisella tetracycline-regulated promoter (FTRp), and transcription was initiated with addition of anhydrotetracycline (ATc), which binds TetR and alleviates TetR association with tetO. Expression levels measured by luminescence correlated with ATc inducer concentrations ranging from 20 to 250 ng ml(-1). In the absence of ATc, luminescence was below the level of detection. The inducible system was also functional during the infection of J774A.1 macrophages, as determined by both luminescence and rescue of a mutant strain with an intracellular growth defect. The repressible system consists of FTRp regulated by a reverse TetR mutant (revTetR), TetR r1.7. Using this system with the lux reporter, the addition of ATc resulted in decreased luminescence, while in the absence of ATc the level of luminescence was not significantly different from that of a construct lacking TetR r1.7. Utilizing both systems, the essentiality of SecA, the protein translocase ATPase, was confirmed, establishing that they can effectively regulate gene expression. These two systems will be invaluable in exploring F. tularensis protein function.
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