Fluorescent Protein Approaches in Alpha Herpesvirus Research. Author Ian Hogue, Jens Bosse, Esteban Engel, Julian Scherer, Jiun-Ruey Hu, Tony Del Rio, Lynn Enquist Publication Year 2015 Type Journal Article 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. Keywords Recombinant Fusion Proteins, Luminescent Proteins, Host-Pathogen Interactions, Alphaherpesvirinae, Biomedical Research, Staining and Labeling, Virology Journal Viruses Volume 7 Issue 11 Pages 5933-61 Date Published 2015 Nov 19 ISSN Number 1999-4915 DOI 10.3390/v7112915 Alternate Journal Viruses PMCID PMC4664988 PMID 26610544 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML