Department of Chemistry, Center for NanoScience (CeNS) and Center for Integrated Protein Science, Munich (CIPSM), Ludwig-Maximilians-Universität München, Butenandtstr. 5-13, D-81377 München, Germany.
Viruses. 2012 May;4(5):777-99. doi: 10.3390/v4050777. Epub 2012 May 4.
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 recruitment of Gag and RNA to the plasma membrane, virion assembly at the membrane and the recruitment of cellular proteins involved in HIV-1 release to the nascent budding site.
荧光方法学的进步使得以前所未有的细节研究生物系统成为可能。在过去的几年中，定量活细胞成像越来越多地被用于研究病毒与细胞的动态相互作用，预计在未来将变得更加不可或缺。在这里，我们描述了用于标记 HIV-1 进行活细胞成像的不同荧光标记策略，以及用于可视化病毒-细胞相互作用各个方面的荧光方法。本综述介绍了定量显微镜在阐明 HIV-1 复制周期晚期动力学方面的实验方法和最近的实验的概述。这包括主要结构蛋白 Gag 与其自身和病毒 RNA 基因组的胞质相互作用，Gag 和 RNA 向质膜的募集，病毒组装在膜上以及涉及 HIV-1 释放的细胞蛋白募集到新生出芽部位。
