口蹄疫病毒非结构蛋白3AB基因的表达及反应原性鉴定  

Prokaryotic Expression and Reactogenicity Identification of Nonstructural Protein 3AB Gene from Foot-and-Mouth Disease Virus

作　　者：孙靖[1,2,3] 向华[3] 靳野[1] 蔡建平[3] 王贵平[3] 尚佑军[1] 宣华[2,3] 刘湘涛[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点实验室国家口蹄疫参考实验室,甘肃兰州730046 [2]吉林大学畜牧兽医学院,吉林长春130062 [3]广东省农业科学院兽医研究所,广东广州510640

出　　处：《安徽农业科学》2009年第14期6372-6374,共3页Journal of Anhui Agricultural Sciences

基　　金：国家"十一五"科技支撑计划项目(2006BAD06A17);广东省科技攻关项目(2005A20902002);广东省社会发展计划项目(2006B20801002)

摘　　要：[目的]研究口蹄疫病毒非结构蛋白3AB基因的表达及反应原性鉴定。[方法]以含有FMDV 3ABC基因的质粒为模板,应用RT-PCR技术扩增口蹄疫病毒非结构蛋白3AB基因并进行序列测定,将3AB基因克隆至表达载体pET-28a(+),选取阳性克隆转化BL21感受态用IPTG诱导表达,并进行SDS-PAGE鉴定与W estern-bolt检测分析。[结果]经SDS-PAGE和W estern-bolt分析表明,在大肠杆菌中成功表达了3AB蛋白,表达的目的蛋白能与FMDV阳性血清发生特异性反应。[结论]该项研究为应用以重组FMDV非结构蛋白为抗原建立感染和免疫动物的鉴别诊断方法提供了依据。[ Objective] The aim was to study the prokaryotic expression and reactogenicity identification of nonstructural protein 3AB gene from foot-and-mouth disease virus （FMDV）. [ Method] With the plasmid containing FMDV 3ABC gene as the template, RT-PCR technique was used to amplify FMDV 3AB gene. After sequencing the amplified FMDV 3AB was cloned into the gene expression vector named pET-28a （ ＋ ） , and then transferred to E. coli BL21. It was induced and expressed by IPTG and then made for SDS-PAGE identification and Western-blot detecting analysis. [ Result] Analysis by SDS-PAGE and Western-blot indicated that 3AB protein was successfully expressed in E. coli BL21 and the expressed target protein could be specifically reacted with the positive serum of FMDV. [ Conclusion] This study would provide the basis for establishing the identification and diagnosis method which can distinguish the infected animals from the vaccinated animals with the recombinant FMDV nonstructural protein as the antigen.

关 键 词：FMDV 非结构蛋白 表达 反应原性 

分 类 号：S188[农业科学—农业基础科学]