title: Self-biotinylation and site-specific double labeling of baculovirus using quantum dots for single-virus in-situ tracking.
abstract: 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.
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
authors.forename: Fuxian
authors.initials: F
authors.lastname: Zhang
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Zheng
authors.forename: Shu-Lin
authors.initials: SL
authors.lastname: Liu
authors.forename: Wen
authors.initials: W
authors.lastname: Lu
authors.forename: Zhenfeng
authors.initials: Z
authors.lastname: Zhang
authors.forename: Cuiling
authors.initials: C
authors.lastname: Zhang
authors.forename: Peng
authors.initials: P
authors.lastname: Zhou
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Gang
authors.initials: G
authors.lastname: Long
authors.forename: Zhike
authors.initials: Z
authors.lastname: He
authors.forename: Dai-Wen
authors.initials: DW
authors.lastname: Pang
authors.forename: Qinxue
authors.initials: Q
authors.lastname: Hu
authors.forename: Hanzhong
authors.initials: H
authors.lastname: Wang
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000378:metabolism* / Q000472:pathogenicity; D019921:Biotinylation; D002460:Cell Line; D049452:Green Fluorescent Proteins / Q000378:metabolism; D045663:Quantum Dots; D011995:Recombination, Genetic / Q000235:genetics; D013050:Spectrometry, Fluorescence; D013194:Staining and Labeling; D014759:Viral Envelope Proteins / Q000378:metabolism
chemical_list: D014759:Viral Envelope Proteins; D049452:Green Fluorescent Proteins
keywords: Autographa californica multiple nucleopolyhedrovirus; Double-labeled virus; Long-term tracking; Quantum dot; Self-biotinylated; Single-virus tracking