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Usage of a Bioluminescence Reporter System to Image Promoter Activity During Host Infection

  • Gili Aviv
  • Ohad Gal-MorEmail author
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 1734)

Abstract

Bioluminescence is the process of production and emission of light by a living organism, usually as the by-product of the oxidative enzyme, luciferase. Currently available technology allows for the exploitation of a bioluminescent reporter system to study bacterial gene regulation during rodent infection, in real time, over a large dynamic range. Here we show how this imaging system can be used to study virulence gene regulation during Salmonella enterica infection in the mouse model. To demonstrate this technique we show the ex vivo expression pattern of the gene dksA, encoding a conserved and pleotropic regulator, which plays a key role in Salmonella pathogenicity [1].

Key words

Luciferase Bioluminescence Reporter gene In vivo imaging Transcriptional regulation Host infection 

Notes

Acknowledgments

The research in Gal-Mor lab is supported by a grant number 1096.39.11/2010 from the German-Israeli Foundation for Scientific Research and Development (GIF); by a grant number 999/14 from the Israel Science Foundation (ISF) and by grant number 3-0000-12435 from Infect-ERA and the Chief Scientist’s Bureau in the Israeli Ministry of Health.

References

  1. 1.
    Azriel S, Goren A, Rahav G, Gal-Mor O (2016) The stringent response regulator DksA is required for Salmonella enterica serovar Typhimurium growth in minimal medium, motility, biofilm formation, and intestinal colonization. Infect Immun 84:375–384CrossRefGoogle Scholar
  2. 2.
    Meighen EA, Szittner RB (1992) Multiple repetitive elements and organization of the lux operons of luminescent terrestrial bacteria. J Bacteriol 174:5371–5381CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Bjarnason J, Southward CM, Surette MG (2003) Genomic profiling of iron-responsive genes in Salmonella enterica serovar Typhimurium by high-throughput screening of a random promoter library. J Bacteriol 185:4973–4982CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Meighen EA (1991) Molecular biology of bacterial bioluminescence. Microbiol Rev 55:123–142PubMedPubMedCentralGoogle Scholar
  5. 5.
    Gal-Mor O, Gibson DL, Baluta D et al (2008) A novel secretion pathway of Salmonella enterica acts as an antivirulence modulator during salmonellosis. PLoS Pathog 4:e1000036CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Nguyen M, Rizvi J, Hecht G (2015) Expression of enteropathogenic Escherichia coli map is significantly different than that of other type III secreted effectors in vivo. Infect Immun 83:130–137CrossRefPubMedGoogle Scholar
  7. 7.
    Trcek J, Fuchs TM, Trulzsch K (2010) Analysis of Yersinia enterocolitica invasin expression in vitro and in vivo using a novel luxCDABE reporter system. Microbiology 156:2734–2745CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.The Infectious Diseases Research LaboratorySheba Medical CenterTel-Hashomer, Ramat GanIsrael
  2. 2.Department of Clinical Microbiology and Immunology, Sackler Faculty of MedicineTel Aviv UniversityTel AvivIsrael

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