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为了获得反应原性较好的猪流行性腹泻病毒(PEDV)S1蛋白,利用原核表达系统对S1基因(60~845 bp)进行扩增并构建原核表达质粒,转化至大肠杆菌BL21(DE3),以1 mmol/L IPTG诱导表达4 h,进行SDS-PAGE和Western blot分析。结果显示:成功构建了pET-30A-S1质粒,重组蛋白得到了表达且具有良好的反应原性,能够被兔PEDV多抗血清识别。本研究为进一步建立ELISA检测方法和亚单位疫苗的研制奠定了物质基础。
In order to obtain S1 gene of porcine epidemic diarrhea virus (PEDV) with good reactivity, the S1 gene (60-845 bp) was amplified by prokaryotic expression system and the prokaryotic expression plasmid was constructed and transformed into Escherichia coli BL21 (DE3) The cells were induced with 1 mmol / L IPTG for 4 h and analyzed by SDS-PAGE and Western blot. The results showed that the plasmid pET-30A-S1 was successfully constructed, the recombinant protein was expressed and had good reactivity and could be recognized by the rabbit polyclonal antiserum against PEDV. This study lays the material foundation for the further establishment of ELISA detection methods and the development of subunit vaccines.