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Visualizing hepatitis B virus with biarsenical labelling in living cells
Author(s) -
Sun Shuzhen,
Yan Jingjun,
Xia Chao,
Lin Yuanyuan,
Jiang Xiaorui,
Liu Haojing,
Ren Huanping,
Yan Junwei,
Lin Jusheng,
He Xingxing
Publication year - 2014
Publication title -
liver international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.873
H-Index - 110
eISSN - 1478-3231
pISSN - 1478-3223
DOI - 10.1111/liv.12419
Subject(s) - green fluorescent protein , hepatitis b virus , transfection , fluorescence microscope , fluorescence , virology , biology , virus , cytoplasm , recombinant dna , chemistry , microbiology and biotechnology , cell culture , gene , biochemistry , genetics , physics , quantum mechanics
Background Study on viruses has greatly benefited from visualization of viruses tagged with green fluorescent protein ( GFP ) in living cells. But GFP tag, as a large inserted fragment, is not suitable for labelling Hepatitis B virus ( HBV ) that is a compact virion with limited internal space. Aim To visualize HBV in living cells, we constructed several recombinant HBV fluorescently labelled with biarsenical dye to track the behaviour of HBV in the cytoplasm of infected cells. Methods By mutagenesis, a smaller size tetracysteine ( TC ) tag (C‐C‐P‐G‐C‐C) that could be bound with a biarsenical fluorescent dye was genetically inserted at different cell epitopes of HBV core protein expressed in transfected cells. Result Confocal microscopy and transmission electron microscopy ( TEM ) observations showed that TC ‐tagged core proteins bound with biarsenical dye could specifically fluoresce in cells and be incorporated into nucleocapsid to form fluorescent virions. The recombinant fluorescent HBV virions retained their infectivity as wild‐type ones. Moreover, tracking of fluorescent HBV particles in living cells reveals microtubule‐dependent motility of the intracellular particles. Conclusion To the best of our knowledge, this is the first time to generate fluorescent HBV virions with biarsenical labelling and to visualize their trafficking in living cells. The fluorescent HBV may become one highly valuable tool for further studying detailed dynamic processes of HBV life cycle and interaction of HBV with host in live‐imaging approach.