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期刊论文

Baculovirus surface display of NS3 nonstructural protein of classical swine fever virus

童德文Xin-Gang Xua De-Wen Tonga Ming-Tang Chioub Yao-Ching HsiehdWen-Ling Shihc Ching-Dong Changb Ming-Huei Liaob Yan-Ming Zhanga Hung-Jen Liubc

Journal of Virological Methods 159 (2009) 259-264,-0001,():

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摘要/描述

Classical swine fever virus (CSFV) causes significant losses in the pig industry in many countries. NS3 proteins of CSFV, which include serine protease and RNA helicase/nucleotide triphosphatase (NTPase) activities, are multifunctional proteins involved in polyprotein processing and viral replication. Previous reports showed that NS3 protein can induce apoptosis in host cells that present cytopathic effects (CPE). Baculovirus/insect cell systems are used widely for recombinant protein production. In this study, one recombinant baculovirus BacSC-NS3 expressing histidine-tagged NS3 with the transmembrane domain (TM) and cytoplasmic domain (CTD) derived from baculovirus envelope protein gp64 of baculovirus was constructed. After infection, NS3 was expressed and anchored to the plasma membrane of Sf-9 cells, as demonstrated byWestern blot assay and confocal microscopy. Immunogold electron microscopy demonstrated that the NS3 glycoprotein successfully displayed on the baculoviral envelope. Animal vaccine tests showed that recombinant baculovirus BacSC-NS3 elicited significantly higher NS3 antibody titers in the treatedmousemodels than the control group. This reportdemonstrated the potential of NS3-pseudotyped baculovirus expression of NS3 protein successfully.

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