IBDV VP3结构蛋白在大肠杆菌中的表达与鉴定

Expression and Identification of IBDV VP3 Structural Protein in E.Coli

应用RT-PCR技术从鸡IBDV总RNA中克隆出893bp的VP3基因,并将其进一步克隆于pET-28a载体T7启动子的下游,构建了pETVP3原核表达载体。经IPTG诱导,在其宿主菌BL21(DE3)中成功表达了35.5,33kDa的鸡IBDVVP3蛋白。经SDS-PAGE和Westernblotting分析,VP3表达产物以包涵体形式存在,并与鸡IBDV抗血清特异性反应。

The VP3 gene about 893bp was obtained from the total RNA of the chicken IBDV by RT-PCR, then the VP3 gene was subcloned into the downstream of T7 promoter of the vector pET-28a to construct expression vector of pETVP3. The recombinant plasmid was transformed into in E. coli BL21（DE3） and induced with IPTG. SDS-PAGE and Western blotting analyses showed that the chicken IBDV VP3 protein of 35.5 and 33 kDa were efficiently expressed and mainly existed as inclusion bodies,with specific reactivity to anti-IBDV antibodies.