口蹄疫病毒3AB基因的表达及活性分析  

EXPRESSION AND ACTIVITY ANALYSIS OF FMDV 3AB PROTEIN

作　　者：王生育[1] 孔繁德[1] 颜江华[2] 林祥梅[3] 韩雪清[3] 吴绍强[3] 黄印尧[1] 

机构地区：[1]厦门出入境检验检疫局,福建厦门361012 [2]厦门大学医学院抗癌研究中心,福建厦门361005 [3]中国检验检疫科学研究院动植物检疫研究所,北京100029

出　　处：《检验检疫科学》2007年第5期21-24,共4页Inspection and Quarantine Science

基　　金：国家质检总局科研项目(2007IK039)

摘　　要：通过重叠PCR合成口蹄疫病毒3AB基因,构建原核表达载体pGEX-5X-1/3AB,转化大肠杆菌BL21(DE3),IPTG诱导表达。重组蛋白主要以包涵体的形式表达,将包涵体洗涤溶解后,采用Na2+离子金属螯合亲和层析柱纯化,逐步透析法复性。ELISA实验表明,目的蛋白能与猪口蹄疫阳性血清发生特异性反应。本研究为建立以基因工程产品为抗原、鉴别诊断自然感染和免疫动物的方法提供了技术条件。The 3AB gene of FMDV was assembled by overlapping PCR. The expression vector pGEX-5X-1/3AB was constructed and transformed into E.coli BL21 （DE3）. The FMDV 3AB protein was expressed after IPTG induction. Recombinant proteins were expressed mainly as inclusion bodies in the system. After inclusion bodies was washed and dissolved, expressed protein was purified by Ni^2＋-affinity chromatographic column, and refolded by following dialysis. ELISA assay suggested that the purified protein react specifically with FMDV positive serum. This study provided technical support for developing recombinant antigen-based method to distinguish naturally infected animals from vaccinated.

关 键 词：口蹄疫病毒 3AB基因 表达 活性 

分 类 号：S852.65[农业科学—基础兽医学]