高致病性猪繁殖与呼吸综合征病毒(JX0601)NSP2基因主要抗原区域的原核表达与纯化  

Prokaryotic Expression and Purification of NSP2 Major Antigen Region from a Highly Pathogenic PRRSV Strain JX0601

作　　者：李相钊[1] 张志[2] 王菲[1] 吴发兴[2] 路希山[1] 张燕霞[2] 李晓成[2] 单虎[1] 

机构地区：[1]青岛农业大学,山东青岛266109 [2]中国动物卫生与流行病学中心,山东青岛266032

出　　处：《动物医学进展》2008年第10期13-17,共5页Progress In Veterinary Medicine

基　　金：国家科技支撑计划项目资助(2007BAD86B05)

摘　　要：本研究采用PCR方法从高致病性猪繁殖与呼吸综合征病毒(JX0601)质粒扩增得到NSP2基因的主要抗原决定区域dNSP2,克隆目的基因连接到pGEM-TEasy,再将其亚克隆到pET-32a上,构建表达载体pET-dNSP2,然后转化大肠埃希菌BL-21(DE3),分别用不同浓度的IPTG诱导表达,经SDS-PAGE电泳分析,所表达融合蛋白的分子质量约为38.3ku,用HisBindPurificationKit试剂盒纯化蛋白,Westernblot结果表明重组蛋白可被PRRSV阳性血清所识别。本试验为进一步研究PRRSV诊断试剂盒奠定了基础。The major antigen region of NSP2 gene, dNSP2, was successfully amplified by PCR and cloned into pGEM-T Easy vector from a plasmid of highly pathogenic PRRSV （JX0601 strain）. After verified by sequencing,this fragment was subcloned into prokaryotic expression vector pET-32a, which constructed the recombinant expression vector pET-dNSP2. Its expression character was induced under different concentrations of IPTG at 37 ℃ in E. coli BL21（DE3）,and analyzed by SDS-PAGE. The results showed that the recombinant protein was about 38.3 ku when purified by His Bind Purification Kit and could react positively with the swine antiserum of PRRSV by Western blot. This research provided fundamental data on diagnostic kit development of PRRSV.

关 键 词：猪繁殖与呼吸综合征病毒 NSP2基因 原核表达 

分 类 号：S852.659.6[农业科学—基础兽医学]