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猪流行性腹泻病毒(PEDV)是引起猪腹泻的重要病原之一,给世界养猪业带来严重危害。本研究对PEDV分离株JS-2纤突蛋白(S)进行抗原表位分析,选择3个富含表位片段的区域。通过RT-PCR扩增基因序列,克隆入p ET32a(+)载体,转化大肠杆菌BL21(DE3)中并进行诱导表达,SDS-PAGE电泳显示,成功表达了3个重组蛋白,分子质量与预期大小一致。将蛋白进行割胶回收,与油佐剂乳化后免疫小鼠,制备超免疫血清,利用PEDV包被抗原进行ELISA检测,结果免疫小鼠血清D450值达1.0左右,证实筛选制备的重组蛋白具有良好的免疫原性和反应原性。
Porcine epidemic diarrhea virus (PEDV) is one of the important pathogens causing swine diarrhea, which brings serious harm to the world pig industry. In this study, antigenic epitopes of JS-2 spike protein (S) of PEDV isolates were analyzed. Three epitope-rich regions were selected. The gene sequence was amplified by RT-PCR, cloned into p ET32a (+) vector and transformed into E.coli BL21 (DE3) for expression. SDS-PAGE electrophoresis showed that three recombinant proteins were successfully expressed with the expected molecular weight Consistent. The protein was recovered by tapping and emulsified with oil adjuvant to immunize mice to prepare hyperimmune serum. ELISA was performed using PEDV-coated antigen. The results showed that serum D450 of immunized mice reached about 1.0, which confirmed that the recombinant protein prepared by screening had good Immunogenicity and reactivity.