【摘要】 参考GenBank中收录的猪生殖与呼吸综合征病毒(PRRSV)JXA1株Nsp2基因序列,设计了1对引物用于扩增截短的Nsp2(deleting Nsp2,dNsp2)基因,用BamHⅠ+XhoⅠ双酶切后将目的基因插入到原核表达载体pET-32a(+)中,转化大肠杆菌BL21(DE3)进行表达,经SDS-PAGE电泳和Western-blot检测,dNsp2基因得到了高效表达,融合蛋白的分子质量约为36.5 ku,表达产物以可溶性方式存在于表达上清液中,表达量可达菌体总蛋白的30%以上,表达产物能被PRRSV变异株抗体所识别,具有较好的生物学活性。PRRSV变异株dNsp2基因的高效表达为该病毒ELISA检测方法的建立及应用奠定了基础。更多还原

【Abstract】 The specific primers were synthesized according to the sequence of Nsp2 gene from porcine reproductive and respiratory syndrome virus published in GenBank.Then the amplified deleting Nsp2(dNsp2) fragment with XhoⅠ and BamHⅠ sites was cloned into the vector pET-32a(+) and the recombinant fusion protein was expressed in Escherichia coli BL21(DE3).The total mass of the fusion protein with 36.5ku was confirmed by SDS-PAGE and Western-blotting.The protein also existed in the supernatant and contained 30% of the total somatic protein.Western-blotting verified that the recombinant protein could be recognized by the porcine variant strain polyclonal antibody against PRRSV,and it had a greater biological activity.The high level expression of dNsp2 gene laid a basis for the establishment of an ELISA for detection of PRRSV.更多还原