Lampe Marko, Briggs John A G, Endress Thomas, Glass Bärbel, Riegelsberger Stefan, Kräusslich Hans-Georg, Lamb Don C, Bräuchle Christoph, Müller Barbara
Department of Virology, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 324, 69120 Heidelberg, Germany.
Virology. 2007 Mar 30;360(1):92-104. doi: 10.1016/j.virol.2006.10.005. Epub 2006 Nov 9.
Human immunodeficiency virus (HIV) delivers its genome to a host cell through fusion of the viral envelope with a cellular membrane. While the viral and cellular proteins involved in entry have been analyzed in detail, the dynamics of virus-cell fusion are largely unknown. Single virus tracing (SVT) provides the unique opportunity to visualize viral particles in real time allowing direct observation of the dynamics of this stochastic process. For this purpose, we developed a double-coloured HIV derivative carrying a green fluorescent label attached to the viral matrix protein combined with a red label fused to the viral Vpr protein designed to distinguish between complete virions and subviral particles lacking MA after membrane fusion. We present here a detailed characterization of this novel tool together with exemplary live cell imaging studies, demonstrating its suitability for real-time analyses of HIV-cell interaction.
人类免疫缺陷病毒（HIV）通过病毒包膜与细胞膜融合将其基因组传递至宿主细胞。虽然参与病毒进入过程的病毒蛋白和细胞蛋白已得到详细分析，但病毒-细胞融合的动力学在很大程度上仍不清楚。单病毒追踪（SVT）提供了独特的机会来实时可视化病毒颗粒，从而直接观察这一随机过程的动力学。为此，我们开发了一种双色HIV衍生物，其携带附着于病毒基质蛋白的绿色荧光标记，并与融合至病毒Vpr蛋白的红色标记相结合，旨在区分完整病毒体和膜融合后缺乏基质蛋白的亚病毒颗粒。我们在此展示了这种新型工具的详细特性以及示例性的活细胞成像研究，证明了其适用于HIV-细胞相互作用的实时分析。
