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Viruses 2015, 7(11), 5933-5961; doi:10.3390/v7112915

Fluorescent Protein Approaches in Alpha Herpesvirus Research

Department of Molecular Biology & Princeton Neuroscience Institute, Princeton University, Princeton, NJ 08544, USA
Present address: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg 20251, Germany
Present address: Boston Biomedical, Inc., Cambridge, MA 02139, USA
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Author to whom correspondence should be addressed.
Academic Editor: Eric O. Freed
Received: 20 August 2015 / Revised: 12 October 2015 / Accepted: 14 October 2015 / Published: 19 November 2015
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Abstract

In the nearly two decades since the popularization of green fluorescent protein (GFP), fluorescent protein-based methodologies have revolutionized molecular and cell biology, allowing us to literally see biological processes as never before. Naturally, this revolution has extended to virology in general, and to the study of alpha herpesviruses in particular. In this review, we provide a compendium of reported fluorescent protein fusions to herpes simplex virus 1 (HSV-1) and pseudorabies virus (PRV) structural proteins, discuss the underappreciated challenges of fluorescent protein-based approaches in the context of a replicating virus, and describe general strategies and best practices for creating new fluorescent fusions. We compare fluorescent protein methods to alternative approaches, and review two instructive examples of the caveats associated with fluorescent protein fusions, including describing several improved fluorescent capsid fusions in PRV. Finally, we present our future perspectives on the types of powerful experiments these tools now offer. View Full-Text
Keywords: alpha herpesvirus; herpes simplex virus; HSV; pseudorabies virus; fluorescent protein; GFP; fluorescence microscopy; capsid alpha herpesvirus; herpes simplex virus; HSV; pseudorabies virus; fluorescent protein; GFP; fluorescence microscopy; capsid
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Hogue, I.B.; Bosse, J.B.; Engel, E.A.; Scherer, J.; Hu, J.-R.; del Rio, T.; Enquist, L.W. Fluorescent Protein Approaches in Alpha Herpesvirus Research. Viruses 2015, 7, 5933-5961.

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