传染性法氏囊病病毒VP2基因原核表达及抗原性分析

Prokaryotic Expression of VP2 Gene of Infectious Bursal Disease Virus and Antigenicity of Expressed Products

从辽宁省某鸡场分离到一株传染性法氏囊病病毒(IBDV)。以该毒株基因组核酸为模板,应用RT-PCR扩增得到VP2基因,构建表达质粒pET28a-VP2,再将其转化至大肠埃希菌BL21(DE3)用IPTG进行诱导表达。表达产物经SDS-PAGE分析在48ku处出现特异性蛋白条带;经Western blot分析VP2蛋白可以与IBDV阳性血清发生特异性反应。用VP2蛋白制备的油乳剂疫苗免疫接种SPF鸡,2周后体内可以检测到特异性抗体。证明VP2蛋白具有重要的应用价值,为IBDV基因工程亚单位疫苗的研制奠定了基础。

An infectious bursal disease virus（IBDV） strain was isolated from a chicken farm of Liaoning province.The cDNA fragment of protective antigen VP2 gene of the strain was amplified by RT-PCR,subcloned into pMD18-T vector and then into pET28a vector,designated as pET28a-VP2 plasmid,The recombinant plasmid pET28a-VP2 was transformed into competent E.coli BL21（DE3） cells and induced with IPTG.The results of SDS-PAGE indicated that the expressed VP2 protein was about 48 ku;The results of Western blot indicated that the expressed VP2 protein has immunological reactive activity.Using the oil emulsion vaccine which made of the expressed VP2 protein to immunize SPF chicken,the results showed that the specific anti-IBDV antibody could be detected in 2 weeks.The results demonstrated that VP2 protein has important applications and provides a theoretical basis for genetically engineered IBDV subunit vaccine production.