Visualization of the African swine fever virus infection in living cells by incorporation into the virus particle of green fluorescent protein-p54 membrane protein chimera
| Autor: | Bruno Hernáez, Covadonga Alonso, José M. Escribano |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2006 |
| Předmět: |
Cell Survival
viruses β-gal β-galactosidase Recombinant Fusion Proteins Green Fluorescent Proteins Biology Recombinant virus GFP African swine fever virus Virus Article MW molecular weight Green fluorescent protein Viral factory Viral life cycle ORF open reading frame Virology Chlorocebus aethiops Morphogenesis ASFV African swine fever virus Animals Vero Cells MOI multiplicity of infection Viral Structural Proteins Ara-c cytosine β-arabinofuranoside Viral culture biology.organism_classification Fusion protein EGFP enhanced green fluorescent protein African Swine Fever Virus wb western blot Fluorescent tagged virus |
| Zdroj: | Virology Repositorio de Resultados de Investigación del INIA Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria INIA INIA: Repositorio de Resultados de Investigación del INIA |
| ISSN: | 1096-0341 0042-6822 |
| Popis: | Many stages of African swine fever virus infection have not yet been studied in detail. To track the behavior of African swine fever virus (ASFV) in the infected cells in real time, we produced an infectious recombinant ASFV (B54GFP-2) that expresses and incorporates into the virus particle a chimera of the p54 envelope protein fused to the enhanced green fluorescent protein (EGFP). The incorporation of the fusion protein into the virus particle was confirmed immunologically and it was determined that p54-EGFP was fully functional by confirmation that the recombinant virus made normal-sized plaques and presented similar growth curves to the wild-type virus. The tagged virus was visualized as individual fluorescent particles during the first stages of infection and allowed to visualize the infection progression in living cells through the viral life cycle by confocal microscopy. In this work, diverse potential applications of B54GFP-2 to study different aspects of ASFV infection are shown. By using this recombinant virus it was possible to determine the trajectory and speed of intracellular virus movement. Additionally, we have been able to visualize for first time the ASFV factory formation dynamics and the cytophatic effect of the virus in live infected cells. Finally, we have analyzed virus progression along the infection cycle and infected cell death as time-lapse animations. © 2006 Elsevier Inc. All rights reserved. |
| Databáze: | OpenAIRE |
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