| 口蹄疫病毒结构蛋白基因vp1的表达与应用研究 |
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| 引用本文： | 邢继兰,潘洁,陈波,饶忠,尤永进,徐泉兴,刘惠莉.口蹄疫病毒结构蛋白基因vp1的表达与应用研究[J].中国预防兽医学报,2007,29(4):299-302. |
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| 作者姓名： | 邢继兰 潘洁 陈波 饶忠 尤永进 徐泉兴 刘惠莉 |
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| 作者单位： | 上海市农业科学院,畜牧兽医研究所,上海,201106 |
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| 基金项目： | 上海市农业科学院科技发展基金 |
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| 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。
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| 关 键 词： | 口蹄疫病毒 结构蛋白 vp1基因表达 |
| 文章编号： | 1008-0589（2007）04-0299-04 |
| 收稿时间： | 2006-07-24 |
| 修稿时间： | 2006年7月24日 |
Expression and application of foot-and-mouth disease virus vp1 gene |
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| XING Ji-lan,PAN Jie,CHEN Bo,RAO Zhong,YOU Yong-jin,XU Quan-xing,LIU Hui-li.Expression and application of foot-and-mouth disease virus vp1 gene[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(4):299-302. |
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| Authors: | XING Ji-lan PAN Jie CHEN Bo RAO Zhong YOU Yong-jin XU Quan-xing LIU Hui-li |
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| Abstract: | 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 elec- trophoresis,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 indi- rect ELISA based on recombinant VP1 protein was positively correlated with liquid phase blocking ELISA,but with no signifi- cance.In conclusion,the indirect ELISA using recombinant vp1 as antigen may be useful for monitoring FMDV antibody. |
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| Keywords: | foot-and-mouth disease virus (FMDV) structural protein expression of vpl gene |
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