A method of alphavirus replicon particle titration based on expression of functional replicase/transcriptase
| Autor: | Gerald R. Kovacs, Timothy J. Zamb, Jacek Kowalski, Seema S Gangolli, Nikolaos Vasilakis |
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| Rok vydání: | 2003 |
| Předmět: |
animal structures
viruses Green Fluorescent Proteins RNA-dependent RNA polymerase Vaccinia virus Alphavirus Vectors in gene therapy medicine.disease_cause complex mixtures Virus Green fluorescent protein Encephalitis Virus Venezuelan Equine Virology Chlorocebus aethiops medicine Animals Replicon Vero Cells biology Defective Viruses DNA-Directed RNA Polymerases biology.organism_classification Molecular biology Luminescent Proteins Togaviridae Venezuelan equine encephalitis virus |
| Zdroj: | Journal of Virological Methods. 109:133-138 |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/s0166-0934(03)00063-6 |
| Popis: | Alphavirus replicon particles are being exploited for a variety of purposes both in vitro as gene expression vectors, and in vivo as vaccines or gene therapy vectors. There is a need for a simple and universal method of titration of replicon particles that is independent of expression of the foreign protein. We devised a method that uses modified vaccinia virus Ankara (MVA) as an indicator virus, to deliver a Venezuelan equine encephalitis virus (VEE) defective helper RNA encoding green fluorescent protein (GFP). Co-infection of cells with the MVA-based indicator and Venezuelan equine encephalitis virus replicon particles (VRP) results in expression of the GFP gene. VRP titer is readily determined by counting fluorescent cells. |
| Databáze: | OpenAIRE |
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