1. Prediction of G gene epitopes of viral hemorrhagic septicemia virus and eukaryotic expression of major antigen determinant sequence.
- Author
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Sun T, Yin WL, Fang BH, Wang Q, Liang CZ, and Yue ZQ
- Subjects
- Amino Acid Sequence, Animals, Antigens, Viral immunology, Cell Line, DNA, Recombinant metabolism, Gene Expression, Plasmids metabolism, Protein Sorting Signals, Transfection, Viral Proteins chemistry, Viral Proteins genetics, Antigens, Viral chemistry, Antigens, Viral genetics, Epitopes chemistry, Genes, Viral, Novirhabdovirus genetics
- Abstract
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.
- Published
- 2017
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