School of Life Science, Beijing Institute of Technology, Beijing 100081, China.
Anal Chem. 2013 May 21;85(10):5263-70. doi: 10.1021/ac4008144. Epub 2013 May 9.
An alternative method for labeling fully replicative enveloped viruses was developed, in which both the biosynthesis and metabolic incorporation of phospholipids in host cells were simultaneously utilized to introduce an azide group to the envelope of the vaccinia virus by taking advantage of the host-derived lipid membrane formation mechanism. Such an azide group could be subsequently used to fluorescently label the envelope of the virus via a bioorthogonal reaction. Furthermore, simultaneous dual-labeling of the virus through the virus replication was realized skillfully by coupling this envelope labeling strategy with "replication-intercalation labeling" of viral nucleic acid. For the first time, it is by natural propagation of the virus in its host cells in the presence of fluorophores that simultaneous dual-labeling of living viruses can be mildly realized with high efficiency in facile and mild conditions.
一种用于标记完全复制包膜病毒的替代方法已经被开发出来，该方法同时利用宿主细胞中磷脂的生物合成和代谢掺入，利用宿主来源的脂质膜形成机制，将叠氮基团引入痘苗病毒的包膜中。通过生物正交反应，这种叠氮基团可以随后被用来荧光标记病毒的包膜。此外，通过将这种包膜标记策略与病毒核酸的“复制嵌入标记”相结合，巧妙地实现了病毒的同时双重标记。这是首次在荧光染料存在的情况下，通过病毒在宿主细胞中的自然繁殖，温和、高效地在简单温和的条件下实现了活病毒的同时双重标记。
