口蹄疫病毒结构蛋白基因vp1的表达与应用研究  

Expression and application of foot-and-mouth disease virus vp1 gene

作　　者：邢继兰[1] 潘洁[1] 陈波[1] 饶忠[1] 尤永进[1] 徐泉兴[1] 刘惠莉[1] 

机构地区：[1]上海市农业科学院畜牧兽医研究所,上海201106

出　　处：《中国预防兽医学报》2007年第4期299-302,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：上海市农业科学院发展基金项目资助

摘　　要：体外克隆口蹄疫病毒vp1基因,构建重组表达载体pET28a-vp1。将此重组质粒转化到受体菌BL21(DE3)中,进行诱导表达,SDS-PAGE和蛋白质印迹分析表明,诱导5h后表达量达到最高,表达产物大小约为33Ku～40Ku,表达蛋白能与口蹄疫病毒阳性血清产生特异性免疫反应。经HPLC纯化后,以重组蛋白为抗原,建立检测VP1蛋白抗体的ELISA方法,检测猪牛血清样品,免疫抗体检测结果与口蹄疫液相阻断ELISA检测结果呈正相关,能反映出免疫抗体动态变化,对临床样品口蹄疫病毒血清抗体检测,两种方法有一定相关性,但不显著。所以以重组VP1蛋白为检测抗原的ELISA方法有望用于口蹄疫免疫抗体监测。Structural protein of （Foot-and-mouth disense, FMDV） vp1 gene was amplified by PCR and cloned into pET28a（＋） vector. The recombinant plasmid pETVP1 was transformed into BL21 （DE3） E.coli strain and the protein expression was induced by IPTG. Expression of protein was examined and identified by SDS-PAGE, Western blot and Enzyme-linked immunosorbent assay （ELISA）. The results showed that VP1 protein expressed in E.coli had a molecular weight of about 33 Ku in SDS-PAGE electrophoresis, and showed immunoreaction with FMDV positive serum. Enzyme linked immunosorbent assay （ELISA） method was established by using recombinant VP1 protein as antigen for detection of FMDV vaccine antibody. Results showed that indirect ELISA based on recombinant VP1 protein was positively correlated with liquid phase blocking ELISA, but with no significance. In conclusion, the indirect ELISA using recombinant vp1 as antigen may be useful for monitoring FMDV antibody.

关 键 词：口蹄疫病毒 结构蛋白 vp1基因表达 

分 类 号：S852.695.6[农业科学—基础兽医学] Q786[农业科学—兽医学]