Self-biotinylation and site-specific double labeling of baculovirus using quantum dots for single-virus in-situ tracking
Self-biotinylation and site-specific double labeling of baculovirus using quantum dots for single-virus in-situ tracking
复制标题
DOI：
10.1016/j.biomaterials.2013.06.030
复制
发表时间：
2013-10-01
期刊：
影响因子：
14
通讯作者：
Wang, Hanzhong
中科院分区：
文献类型：
--
作者：
Zhang, Fuxian;Zheng, Zhenhua;Wang, Hanzhong
单病毒标记和追踪是研究病毒细胞相互作用的有力工具。本文以杆状病毒为模型，发展了一种标记病毒包膜和病毒衣壳的生物化学方法。在病毒复制过程中，AcMNPV的病毒囊膜被生物素标记，并与量子点（QD）结合，而病毒核衣壳则被绿色荧光蛋白（GFP）标记。建立的标记程序不影响病毒的感染性，表明双标记的病毒保留了功能结构，并可以跟踪病毒在宿主细胞中的定位和移动。双标记的病毒还显示出用于原位和实时可视化单个病毒颗粒内化到宿主细胞中的潜力。此外，病毒包膜和病毒核衣壳的解体过程可以长时间监测（长达2小时）。利用所建立的方法，已经揭示了标记的杆状病毒和宿主细胞之间的一些相互作用的细节。该方法具有高效、特异、简便和保持病毒感染性等优点，为研究病毒与细胞的相互作用提供了一种新的手段。(C)2013爱思唯尔有限公司版权所有。
Single-virus labeling and tracking represent a powerful tool to study virus cell interactions. Using baculovirus as a model, here we developed a biochemical method for labeling both the viral envelope and the viral capsid of a virus. Viral envelope of the baculovirus AcMNPV was self-biotinylated and site-specifically conjugated with quantum dots (QDs) following one-step binding reaction, while the viral nucleocapsid was site-specifically labeled with green fluorescent protein (GFP) during viral replication. The established procedure of labeling did not affect viral infectivity, showing that the double-labeled virus retained functional structure and could be tracked for viral localization and movement in the host cells. The double-labeled virus also demonstrated the potential to be used for in-situ and real-time visualizing the internalization of a single viral particle into the host cells. Furthermore, the disassembly processes of the viral envelope and the viral nucleocapsid could be monitored for a long period of time (up to 2 h). Using the established method, several interaction details between the labeled baculoviruses and the host cells have been revealed. Given its advantages in high efficiency, high specificity, convenience and the maintenance of viral infectivity, the established approach provides a promising means for elucidating virus cell interactions. (C) 2013 Elsevier Ltd. All rights reserved.