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Updated: Aug 6, 2026
Methodology for the Efficient Generation of Fluorescently Tagged Vaccinia Virus Proteins
Published on: January 17, 2014
Propagation of fluorescent viruses in growing plaques
Luis J Alvarez1, Philippe Thomen, Tatyana Makushok
1Laboratoire de Physique Statistique, Ecole Normale Supérieure, Paris, France.
Abstract:
To study virus propagation, we have developed a method by which the propagation of the Lambda bacteriophage can be observed and quantified. This is done by creating a fusion protein of the capsid protein gpD and the enhanced yellow fluorescent protein (EYFP). We show that this fusion allows capsid formation and that the modified viruses propagate on a surface covered with host bacteria thus forming fluorescent plaques. The intensity of fluorescence in a growing plaque determines the distribution of phages. This provides a new tool to study the propagation of infection at the microscopic level.
Insights
Researchers developed a new method to observe virus propagation using a fluorescent bacteriophage. This technique allows for quantifying viral spread and infection distribution at the microscopic level.
Area of Science:
- Microbiology
- Virology
- Biotechnology
Background:
- Studying virus propagation is crucial for understanding infection dynamics.
- Existing methods for observing bacteriophage propagation have limitations in real-time quantification.
- Lambda bacteriophage is a model organism for studying viral infection mechanisms.
Purpose of the Study:
- To develop a novel method for observing and quantifying Lambda bacteriophage propagation.
- To create a tool for visualizing viral spread at the microscopic level.
Main Methods:
- Engineered a fusion protein combining the Lambda bacteriophage capsid protein gpD with enhanced yellow fluorescent protein (EYFP).
- Verified that the fusion protein allows for proper capsid formation.
- Demonstrated virus propagation on a surface with host bacteria, resulting in fluorescent plaques.
Main Results:
- The developed fusion protein enables the formation of functional Lambda bacteriophage capsids.
- Modified bacteriophages successfully propagated on host bacteria, forming visible fluorescent plaques.
- Fluorescence intensity within plaques correlates with phage distribution, providing a quantifiable measure of viral spread.
Conclusions:
- The EYFP-gpD fusion protein serves as an effective tool for visualizing and quantifying bacteriophage propagation.
- This method offers a new approach to study infection dynamics at the microscopic level.
- The fluorescent plaque assay provides insights into phage distribution and infection spread.
