【摘要】 用RT-PCR从感染烟草花叶病毒(Tobacco mosaic virus,TMV)的烟草样品中克隆该病毒的外壳蛋白基因,病毒外壳蛋白基因再亚克隆到原核表达载体pET-30a(+)中,构建成重组原核表达载体,转化大肠杆菌BL21,经IPTG诱导、Ni+NTA亲和柱纯化获重组蛋白。以纯化的重组蛋白为抗原免疫家兔,制备TMV外壳蛋白多克隆抗体,并用制备的多克隆抗体建立了更简便、经济、特异性更强的DOT-ELISA检测法。更多还原

【Abstract】 The full length cDNA of tobacco mosaic virus( TMV) which encoded coat protein was cloned from the virus infected tobacco samples by RT-PCR,and subcloned into a prokaryotic expression vector pET-30a( +). The recombinant prokaryotic expression vector was used to transform Escherichia coli BL21. With induction of IPTG and purification of Ni + NTA affinity column the purified recombinant protein was used to immunize rabbits for production of polyclonal antibodies against the coat protein of TMV. Using polyclonal antibodies and DOTELISA were established for reliable,sensitive and specific detection of TMV.更多还原