Visualization of herpes simplex virus type 1 virions using fluorescent colors
| Autor: | Poorval Joshi, Prashant Desai, Laura Dingle, Lyns Etienne, Peter Grzesik, Eugene Huang |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cerulean viruses Recombinant Fusion Proteins Green Fluorescent Proteins Color Herpesvirus 1 Human Biology medicine.disease_cause Protein Engineering Virus Replication Virus Article Green fluorescent protein Cell Line 03 medical and health sciences Virology Chlorocebus aethiops medicine Animals Gene Vero Cells Fluorescent Dyes Cell Membrane Luminescent Proteins 030104 developmental biology Herpes simplex virus Capsid Capsid Proteins mCherry Fluorescent tag |
| Popis: | Our laboratory was one of the first to engineer a live fluorescent tag, enhanced green fluorescent protein (eGFP), that marked the capsid of herpes simplex virus type 1 (HSV-1) and subsequently maturing virus as the particle made its way to the cell surface. In the present study we sought to increase the repertoire of colors available as fusion to the small capsid protein, VP26, so that they can be used alone or in conjunction with other fluorescent tags (fused to other HSV proteins) to follow the virus as it enters and replicates within the cell. We have now generated viruses expressing VP26 fusions with Cerulean, Venus, mOrange, tdTomato, mCherry, and Dronpa3 fluorescent proteins. These fusions were made in a repaired UL35 gene (VP26) background. These fusions do not affect the replication properties of the virus expressing the fusion polypeptide and the fusion tag was stably associated with intranuclear capsids and mature virions. Of note we could not isolate viruses expressing fusions with fluorescent proteins that have a tendency to dimerize. |
| Databáze: | OpenAIRE |
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