%0 Journal Article %T Interaction of Tetraspanin-3 in Fenneropenaeus chinensis with WSSV in vitro %A GUAN,Guangkuo %A LIU,Qinghui %A HUANG,Jie %J 渔业科学进展en %@ 2095-9869 %V 36 %N 6 %D 2015 %P 56-62 %K Tetraspanin-3; Fenneropenaeus chinensis; WSSV; Interaction %X White spot syndrome virus (WSSV) is a widespread and disastrous viral pathogen of cultured shrimp that infects many different species of crustaceans. Tetraspanin-3 protein belongs to the tetraspanin superfamily, which plays an important role in signal transduction and immune process. Increasing evidence has shown that tetraspanins play important roles in virus invasion, penetration and fusion events. Antibody to the large extracellular loop (LEL) of Tetraspanin-3 of Fenneropenaeus chinensis can block the WSSV infection. However, whether the F. chinensis Tetraspanin-3 is involved directly with WSSV remained largely unknown. In this research, we cloned the LEL fragment of Tetraspanin-3 from shrimp F. chinensis (FcT3L) and ligated it with prokaryotic expression vector pBAD/gⅢA to get recombinant expression vector pBAD/gⅢA-FcT3L. After transformed into E. coli (TOP 10) and induced using the L-arabinose, we successfully obtained the pure recombinant protein using Co2+ affinity chromatography purification. Mass spectrometry analysis showed the correctness of the recombinant protein. Moreover, FcT3L was labeled with DIG and incubated with WSSV and the results showed that FcT3L can interact with VP26. Furthermore, recombinant VP26, VP28N, VP28C and VP37 were separated by SDS-PAGE and transferred to PVDF membranes. After incubating with DIG-labeled FcT3L, a positive band was shown with VP26. To confirm the interaction of FcT3L with VP26, FcT3L was coated with 96-well plates and different amount of VP26 was added. ELISA assay showed that the interaction between FcT3L and VP26 grew stronger with the increasing concentration of FcT3L. Results suggested that VP26 might bind to the Tetraspanin-3 and act to help the WSSV diffuse and penetrate into the cytoplasm. The detailed mechanism needs further investigation. %R 10.11758/yykxjz.20150609 %U http://journal.yykxjz.cn/yykxjzen/ch/reader/view_abstract.aspx %1 JIS Version 3.0.0