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Bioluminescent Bacterial Imaging In Vivo
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In Vivo Bacterial Imaging Using Bioluminescence.

Mariette Barbier1, Justin Bevere1, F Heath Damron2

  • 1Department of Microbiology, Immunology and Cell Biology, West Virginia University School of Medicine, Morgantown, WV, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 3, 2018
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Summary

Bacterial luminescence offers a way to track gene activity noninvasively. This study shows how the lux reporter system can quantify promoter activity in real-time within bacteria, plants, insects, and mice.

Keywords:
BacteriaBacterial pathogenesisBioluminescenceIn vivo imagingPathogen-host interactionPromoter activityReporter system

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Bacterial luminescence provides a powerful tool for real-time monitoring of biological processes.
  • The lux reporter system, utilizing bacterial luciferase, is a well-established method for assessing gene expression.
  • Quantifying promoter activity is crucial for understanding gene regulation in various biological contexts.

Purpose of the Study:

  • To detail the application of the lux reporter system for measuring bacterial promoter activity.
  • To describe the construction of promoter fusions with bacterial luciferase.
  • To demonstrate real-time quantification of promoter activity in diverse in vitro and in vivo models.

Main Methods:

  • Utilizing the pUC18T-mini-Tn7T-lux-Tp vector for constructing promoter-lux fusions.
  • Employing bacterial luminescence for real-time monitoring of promoter activity.
  • Applying the system across plant, insect, and murine infection models for in vivo validation.

Main Results:

  • Successful construction of promoter fusions with bacterial luciferase.
  • Demonstrated real-time quantification of promoter activity.
  • Validation of the lux reporter system in multiple complex biological systems.

Conclusions:

  • The lux reporter system, when integrated with the pUC18T-mini-Tn7T-lux-Tp vector, is effective for noninvasive, continuous monitoring of bacterial promoter activity.
  • This methodology enables robust real-time quantification of gene expression in various in vitro and in vivo settings.
  • The system's versatility makes it valuable for studying bacterial gene regulation in diverse model organisms and infection scenarios.