| A型口蹄疫病毒结构蛋白VP1的表达·纯化及活性检测(英文) |
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| 引用本文： | 李菁,林彤,高闪电,丛国正,独军政,邵军军,常惠芸.A型口蹄疫病毒结构蛋白VP1的表达·纯化及活性检测(英文)[J].农业科学与技术,2010,11(2). |
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| 作者姓名： | 李菁 林彤 高闪电 丛国正 独军政 邵军军 常惠芸 |
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| 作者单位： | 李菁,林彤,邵军军,常惠芸,LI Jing,LIN Tong,SHAO Jun-jun,CHANG Hui-yun(中国农业科学院兰州兽医研究所国家口蹄疫参考实验室,甘肃兰州,730046);高闪电,丛国正,GAO Shan-dian,CONG Guo-zheng(中国农业科学院兰州兽医研究所农业部畜禽病毒学重点开放实验室,甘肃兰州,730046);独军政,DU Jun-zheng(中国农业科学院兰州兽医研究所家畜疾病病原生物学国家重点开放实验室,甘肃兰州,730046) |
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| 基金项目： | 国家科技支撑计划，转基因重大专项 |
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| 摘 要： | 目的]表达口蹄疫病毒结构蛋白VP1,并对其进行纯化和相关活性的检测。方法]以A型口蹄疫病毒结构蛋白VP1重组质粒pGEM-VP1为模板,用特异性表达引物扩增其编码区,经酶切后与pGEX-4T-1、pPROExHTb等原核表达载体相连,转化大肠杆菌BL21(DE3)pLysS表达菌株,经IPTG诱导表达,以实现VP1蛋白的表达,SDS-PAGE鉴定融合蛋白的表达并对2种载体重组菌进行超声裂解,取上清和沉淀电泳,用金属螯合亲合层析法对His-VP1进行纯化。结果]SDS-PAGE电泳分析显示pPRO-VP1诱导后目的条带为33Ku,与预期结果相符;目的蛋白纯化后,在电泳图片上显示较为清晰的单一条带;His-VP1可与A型口蹄疫病毒豚鼠灭活阳性血清发生特异性的反应。结论]表达及纯化了具有高特异性的A型口蹄疫病毒结构蛋白VP1,为深入研究结构蛋白VP1在口蹄疫病毒致病机制中的作用奠定了基础。
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| 关 键 词： | A型口蹄疫病毒 结构蛋白VP1 表达及纯化 |
Expression, Purification and Activity Detection of VP1 of A-type FMDV |
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| LI Jing,LIN Tong,GAO Shan-dian,CONG Guo-zheng,DU Jun-zheng,SHAO Jun-jun,CHANG Hui-yun.Expression, Purification and Activity Detection of VP1 of A-type FMDV[J].Agricultural Science ＆ Technology,2010,11(2). |
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| Authors: | LI Jing LIN Tong GAO Shan-dian CONG Guo-zheng DU Jun-zheng SHAO Jun-jun CHANG Hui-yun |
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| Institution: | 1.National Foot-and-mouth Disease Reference Laboratory; Lanzhou Veterinary Research Institute; Chinese Academy of Agricultural Sciences; Lanzhou; 730046; 2.Chinese Academy of Agricultural Sciences; Key Laboratory of Animal Virology of Ministry of Agriculture; Lanzhou 730046; 3.Chinese Academy of Agricultural Sciences; State Key Laboratory of Veterina...; |
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| Abstract: | Objective] The research aimed to induce the expression of FMDV structural protein VP1 in E.coli and purify the protein,then detect the activity.Method] The fragment coding VP1 was amplified by PCR and doubly digested with BamH Ⅰ and XhoⅠ,then cloned into expression vector pGEX-4T-1 and pPROExHTb respectively to get recombinant plasmid pGEX-4T-1-VP1 and pPROExHTb-VP1.The recombinant plasmid pGEX-4T-1-VP1 and pPROExHTb-VP1 was transformed into E.coli BL21(DE3)and induced by IPTG,fusion protein was identifie... |
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| Keywords: | A-type FMDV Structural protein VP1 Expression and purification |
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