关键词: 牛病毒性腹泻病毒, E2基因, E2蛋白,  , 原核表达, 多克隆抗体

Abstract: Using prokaryotic system to express bovine viral diarrhea virus (BVDV) E2 protein, we prepared mouse polyclonal antibody. RNA was extracted by MDBK cell proliferation virus, the fulllength E2 gene was amplified by RTPCR, truncated primers were designed to construct the optimal expression vector pET-30-E2 and transformed BL21 after bioinformatics analysis. The protein expression was induced by IPTG, the protein expression was analyzed by SDSPAGE electrophoresis, and mice were immunized with purified protein and Freund’s adjuvant. Enzyme linked immunosorbent assay (ELISA) was used to determinate the antibody titer level, western blotting (WB) and immuno fluorescence assay (IFA) were used to ver
ify the specificity of the antibody. The results show that E2 gene was successfully expressed in E.coli, the protein size is 32 000 and it
is insoluble. The protein content is 0.4 mg/mL, the titer of polyclonal antibody is 1∶256 000, and it can specifically bind to the recombin
ant protein and virus in cells.

Key words: Bovine viral diarrhea virus, E2 gene,  , E2 protein, prokaryotic expression, polyclonal antibody