Quantitative Live-Cell Imaging of Human Immunodeficiency Virus (HIV-1) Assembly 英文参考文献.docVIP

Viruses 2012, 4, 777-799; doi:10.3390/v4050777 OPEN ACCESS viruses ISSN 1999-4915 /journal/viruses Review Quantitative Live-Cell Imaging of Human Immunodeficiency Virus (HIV-1) Assembly Viola Baumg?rtel 1, Barbara Müller 2,* and Don C. Lamb 1,3,* 1 Department of Chemistry, Center for NanoScience (CeNS) and Center for Integrated Protein Science, Munich (CIPSM), Ludwig-Maximilians-Universit?t München, Butenandtstr. 5-13, D-81377 München, Germany; E-Mail: viola.baumgaertel@cup.uni-muenchen.de 2 Department of Infectious Diseases, Virology, University Hospital Heidelberg, Im Neuenheimer Feld 324, D-69120 Heidelberg, Germany 3 Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61820, USA * Authors to whom correspondence should be addressed; E-Mails: barbara.mueller@med.uni-heidelberg.de (B.M.); d.lamb@lmu.de (D.C.L.); Tel.: +49-6221-56-1325 (B.M.); +49-89-2180-77564 (D.C.L.); Fax: +49-6221-56-5003 (B.M.); +49-89-2180-77560 (D.C.L.). Received: 26 March 2012 / Accepted: 24 April 2012 / Published: 4 May 2012 Abstract: Advances in fluorescence methodologies make it possible to investigate biological systems in unprecedented detail. Over the last few years, quantitative live-cell imaging has increasingly been used to study the dynamic interactions of viruses with cells and is expected to become even more indispensable in the future. Here, we describe different fluorescence labeling strategies that have been used to label HIV-1 for live cell imaging and the fluorescence based methods used to visualize individual aspects of virus-cell interactions. This review presents an overview of experimental methods and recent experiments that have employed quantitative microscopy in order to elucidate the dynamics of late stages in the HIV-1 replication cycle. This includes cytosolic interactions of the main structural protein, Gag, with itself and the viral RNA genome, the recr