Fusion of a fluorescent protein to the pUL25 minor capsid protein of pseudorabies virus allows live-cell capsid imaging with negligible impact on infection

Authors

Kevin P. Bohannon, Northwestern University Feinberg School of Medicine
Patricia J. Sollars, University of Nebraska-LincolnFollow
Gary E. Pickard, University of Nebraska-LincolnFollow
Gregory A. Smith, Northwestern University Feinberg School of MedicineFollow

Date of this Version

1-2012

Citation

J Gen Virol. 93(Pt 1): pp. 124-129, DOI: 10.1099/vir.0.036145-0.

Comments

Copyright © 2012 SGM. Used by permission.

Abstract

In order to resolve the location and activity of submicroscopic viruses in living cells, viral proteins are often fused to fluorescent proteins (FPs) and visualized by microscopy. In this study, we describe the fusion of FPs to three proteins of pseudorabies virus (PRV) that allowed imaging of capsids in living cells. Included in this study are the first recombinant PRV strains expressing FP–pUL25 fusions based on a design applied to herpes simplex virus type 1 by Homa and colleagues. The properties of each reporter virus were compared in both in vitro and in vivo infection models. PRV strains expressing FP–pUL25 and FP–pUL36 preserved wild-type properties better than traditional FP–pUL35 isolates in assays of plaque size and virulence in mice. The utility of these strains in studies of axon transport, nuclear dynamics and viral particle composition are documented.