@article{Sun_Yin_Fang_Wang_Liang_Yue_2017, title={Prediction of G gene epitopes of viral hemorrhagic septicemia virus and eukaryotic expression of major antigen determinant sequence}, volume={63}, url={https://cellmolbiol.org/index.php/CMB/article/view/1638}, DOI={10.14715/cmb/2017.63.7.14}, abstractNote={This study aims to express fish Viral hemorrhagic septicemia virus (VHSV) G main antigen domain by using Bac-to-bac expression system. Using bioinformatics tools, B cell epitope of VHSV G gene was predicted, and G main antigen domain was optimized. GM gene was inserted into pFastBac1 vector, then transferred recombinant plasmid into DH10Bac to get recombinant rBacmid-GM. Obtained shuttle plasmid rBacmid-GM was transfected into sf9 cells. GM expression was examined using by PCR and western-blot. Results indicated that G main antigen domain gene of VHSV was successfully cloned and sequenced which contains 1209 bp. PCR proved that shuttle plasmid rBacmid-GM was constructed correctly. SDS-PAGE electrophoresis analysis detected a band of protein about 45kD in expression product of G gene. Obtained recombinant G protein reacted with VHSV-positive serum that was substantiated by western-blot analysis. In conclusion, the main antigen domain of VHSV G was successfully expressed in the Bac-to-Bac baculovirus system.}, number={7}, journal={Cellular and Molecular Biology}, author={Sun, T. and Yin, W-L. and Fang, B-H. and Wang, Q. and Liang, C-Z. and Yue, Z-Q.}, year={2017}, month={Aug.}, pages={84–88} }