Bioinformatics analysis and prokaryotic expression in E0 gene of bovine viral diarrhea virus and antigenic detection of the expressed protein

This paper studies the protein bioinformatics and antigenicity in E0 of bovine viral diarrhea virus. The laboratory cloning and sequencing of bovine viral diarrhea virus strain yak E0 gene is analyzed. At the same time, the E0 gene is re-amplified by primers containing restriction sites, start codon, stop codon, which is cloned into pMD18_T vector, and the positive recombinant clone is identified by restriction enzyme digestion, then the E0 is cloned into prokaryotic expression vector PET-28A and transferred into JM109. The positive recombinant is identified by digestion, plasmid PCR, and transferred into rosetta (DE3) strain for inducible expression. SDS-PAGE test results show that gene expression is higher, the expression of fusion protein of was about 33ku, and the western-blotting analysis shows that the purpose of protein has good antigenicity.