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Live-Cell Imaging of the Life Cycle of Bacterial Predator Bdellovibrio bacteriovorus using Time-Lapse Fluorescence Microscopy
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Visualizing Bdellovibrio bacteriovorus by Using the tdTomato Fluorescent Protein
Somdatta Mukherjee1, Kimberly M Brothers2, Robert M Q Shanks2
1Department of Oral Biology, Rutgers School of Dental Medicine, Newark, New Jersey, USA.
Applied and Environmental Microbiology
|December 30, 2015
Summary
Bdellovibrio bacteriovorus, a predatory bacterium, can be tracked in real-time using a new fluorescent reporter plasmid. This innovation simplifies monitoring its growth and interactions with prey bacteria and biofilms.
Area of Science:
- Microbiology
- Bacteriology
- Proteobacteria
Background:
- Bdellovibrio bacteriovorus is a Gram-negative bacterium known for its predatory lifestyle.
- Its potential for controlling pathogenic bacteria and biofilms is gaining attention.
- Current methods for monitoring predation, like plaque analysis, are indirect.
Purpose of the Study:
- To develop a more convenient method for monitoring Bdellovibrio bacteriovorus proliferation.
- To introduce a fluorescent reporter system for real-time tracking of B. bacteriovorus.
Main Methods:
- A plasmid (pMQ414) expressing the tdTomato fluorescent protein was introduced into B. bacteriovorus strains.
- The fluorescent construct was used to monitor predator proliferation under various conditions.
- Visualization was performed in liquid cultures, biofilms, and co-culture with human epithelial cells.
Main Results:
- The tdTomato reporter allowed convenient, real-time monitoring of B. bacteriovorus proliferation.
- Predation dynamics were successfully replicated using fluorescence, validating the method.
- The fluorescent predator was visualized in diverse biological contexts, including biofilms and host cell association.
Conclusions:
- The pMQ414 plasmid provides a valuable tool for real-time monitoring of B. bacteriovorus.
- This fluorescent reporter simplifies the study of B. bacteriovorus predation and interactions.
- The method enhances research into the potential applications of B. bacteriovorus in controlling bacterial infections and biofilms.
