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Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Tracking hantavirus nucleocapsid protein using intracellular antibodies | Li et al. Virology Journal 2010 7 339 http www.virologyj.eom content 7 1 339 VIROLOGY JOURNAL RESEARCH Open Access Tracking hantavirus nucleocapsid protein using intracellular antibodies 2t c 71 1t 11 1 2 Jiandong Li T Quanfu Zhang T Tao Wang Chuan Li Mifang Liang Dexin Li Abstract Background Hantavirus nucleocapsid N protein is a multifunctional viral macromolecule involved in multiple stages of the viral replication cycle. The intracellular trafficking of N protein during virus assembly remains unclear. Methods We used N protein-specific intracellular expressed antibodies to track the localization and distribution of Hantaan virus and Seoul virus N protein. The N protein-specific antibody single-chain variable antibody fragments scFvs which bind an N-terminal linear epitope L13F3 and C-terminal conformational domain H34 were intracellularly expressed in the endoplasmic reticulum ER by fusion of the SEKDEL retention signal peptide at the carboxyl terminus and in the cytoplasm Cyto by deletion of the ER membrane target signal peptide. Stable Vero-E6 cell lines expressing intracellular scFvs were either infected with hantavirus or transfected with an N protein expression plasmid virus replication and N protein intracellular localization were determined. Result N protein co-localized with scFvs in the ER and cytoplasm with or without viral membrane glycoproteins. Hantavirus replication was inhibited in both the scFvs-ER- and scFvs-Cyto-expressing stable cell lines. Conclusion N protein may be expressed in the ER retention signal peptide of KDEL circulating region ER cis-Golgi without the assistance of G protein and so expression of N protein in both the cytoplasm and within the ER cis-Golgi plays an important role in virus replication. 1. Background Hantaviruses are members of the Bunyaviridae family which contain three negative-sense single-stranded RNA genome segments designated large L medium M and small S 1 . The S M and L segments encode the nucleocapsid protein
