Huang Cheng-Yen, Chang Wen
Academia Sinica, Institute of Molecular Biology, Taipei, Taiwan, ROC.
Methods Mol Biol. 2012;890:123-33. doi: 10.1007/978-1-61779-876-4_7.
The recently developed technique of live cell imaging has found numerous applications, including the detection of virus movements in living cells. To monitor virus motility, viruses or cellular proteins are fused with fluorescence markers and then detected by time-lapse fluorescence microscopy. These techniques allow kinetic analyses of individual virus particles in motion during the virus entry process as well as monitoring of dynamic interactions between viruses and cellular structures in real time. The methods presented here describe how to construct a fluorescent recombinant vaccinia virus expressing the core protein A4L fused to mCherry, and how to detect the virus movement on actin-EYFP-expressed HeLa cells.
最近开发的活细胞成像技术已得到广泛应用，包括检测活细胞中的病毒运动。为了监测病毒的运动性，将病毒或细胞蛋白与荧光标记物融合，然后通过延时荧光显微镜进行检测。这些技术能够对病毒进入过程中单个运动的病毒颗粒进行动力学分析，并实时监测病毒与细胞结构之间的动态相互作用。本文介绍的方法描述了如何构建表达与mCherry融合的核心蛋白A4L的荧光重组痘苗病毒，以及如何检测在表达肌动蛋白-EYFP的HeLa细胞上病毒的运动。
