Construction and characterization of a fluorescently labeled infectious human immunodeficiency virus type 1 derivative

Journal of Virology
Barbara MüllerHans-Georg Kräusslich

Abstract

The introduction of a label which can be detected in living cells opens new possibilities for the direct analysis of dynamic processes in virus replication, such as the transport and assembly of structural proteins. Our aim was to generate a tool for the analysis of the trafficking of the main structural protein of human immunodeficiency virus type 1 (HIV-1), Gag, as well as for the analysis of virus-host cell interactions in an authentic setting. We describe here the construction and characterization of infectious HIV derivatives carrying a label within the Gag polyprotein. Based on our initial finding that a short epitope tag could be inserted near the C terminus of the matrix domain of Gag without affecting viral replication, we constructed HIV derivatives carrying the egfp gene at the analogous position, resulting in the expression of a Gag-EGFP fusion protein in the authentic viral context. Particles displaying normal viral protein compositions were released from transfected cells, and Gag-EGFP was efficiently processed by the viral protease, yielding the expected products. Furthermore, particles with mature morphology were observed by thin-section electron microscopy. The modified virus was even found to be infectious, al...Continue Reading

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Citations

Apr 11, 2008·European Biophysics Journal : EBJ·Thomas EndressDon C Lamb
Feb 1, 2007·Virus Genes·Evan Burkala, Mary Poss
Mar 5, 2011·Journal of Neuroimmune Pharmacology : the Official Journal of the Society on NeuroImmune Pharmacology·Kate L JonesJohnson Mak
Dec 18, 2012·Current HIV/AIDS Reports·Anupriya AggarwalStuart G Turville
Aug 7, 2013·Trends in Microbiology·Barbara Müller, Mike Heilemann
Jun 10, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Nadia RuthardtChristoph Bräuchle
Mar 12, 2011·Nature Cell Biology·Viola BaumgärtelDon C Lamb
Dec 7, 2007·Nature Methods·Stuart G TurvilleMelissa Robbiani
Sep 23, 2006·Nature Methods·Nathalie ArhelPierre Charneau
May 16, 2012·Proceedings of the National Academy of Sciences of the United States of America·Mickaël LelekChristophe Zimmer
May 20, 2009·The Journal of Biological Chemistry·Dorothée MolleEdouard Bertrand
Aug 12, 2009·The Journal of Biological Chemistry·Barbara MüllerHans-Georg Kräusslich
Jun 26, 2009·The Journal of Biological Chemistry·Maier LorizateHans-Georg Kräusslich
Oct 21, 2011·Human Gene Therapy·Christine VoelkelChristopher Baum
Sep 30, 2010·Antimicrobial Agents and Chemotherapy·Stephan KrembRuth Brack-Werner
Oct 1, 2011·Journal of Virology·Christine E EngelandHans-Georg Kräusslich
Dec 30, 2011·Journal of Virology·Herwig KoppensteinerMichael Schindler
Apr 20, 2011·The Journal of Veterinary Medical Science·Hsu-Chung Gabriel ChenShiow-Suey Lai
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Feb 25, 2011·PloS One·Cândida F PereiraJohnson Mak
Dec 5, 2012·PloS One·Jonas HelmaHeinrich Leonhardt
Mar 29, 2008·PLoS Pathogens·Karine GoussetEric O Freed
Nov 7, 2009·PLoS Pathogens·Sergey IvanchenkoDon C Lamb

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