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Journal of Virology, May 2003, p. 6070-6075, Vol. 77, No. 10
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.10.6070-6075.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Green Fluorescent Protein-Tagged Retroviral Envelope Protein for Analysis of Virus-Cell Interactions

Dirk Spitzer,1,{dagger} Kurt E. J. Dittmar,1 Manfred Rohde,2 Hansjörg Hauser,1* and Dagmar Wirth1

Department of Gene Regulation and Differentiation,1 Department of Microbial Pathogenesis and Vaccine Research, German Research Center for Biotechnology, D-38124 Braunschweig, Germany2

Received 30 April 2002/ Accepted 11 February 2003

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.

* Corresponding author. Mailing address: Department of Gene Regulation and Differentiation, German Research Center for Biotechnology, D-38124 Braunschweig, Germany. Phone: 531 6181 250. Fax: 531 6181 262. E-mail: hha{at}gbf.de.

{dagger} Present address: Division of Rheumatology, Washington University School of Medicine, St. Louis, MO 63110.

Journal of Virology, May 2003, p. 6070-6075, Vol. 77, No. 10
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.10.6070-6075.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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