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Green fluorescent protein-tagged retroviral envelope protein for analysis of virus-cell interactions
Dirk Spitzer1, Kurt E J Dittmar, Manfred Rohde
1Department of Gene Regulation and Differentiation, German Research Center for Biotechnology, D-38124 Braunschweig, Germany.
Abstract:
Fluorescent retroviral envelope (Env) proteins were developed for direct visualization of viral particles. By fusing the enhanced green fluorescent protein (eGFP) to the N terminus of the amphotropic 4070A envelope protein, extracellular presentation of eGFP was achieved. Viruses incorporated the modified Env protein and efficiently infected cells. We used the GFP-tagged viruses for staining retrovirus receptor-positive cells, thereby circumventing indirect labeling techniques. By generating cells which conditionally expressed the GFP-tagged Env protein, we could confirm an inverse correlation between retroviral Env expression and infectivity (superinfection). eGFP-tagged virus particles are suitable for monitoring the dynamics of virus-cell interactions.