数字农科院2.0

伪狂犬病病毒gE基因主要抗原表位区的原核表达及其在疫苗接种和自然感染鉴别诊断中的应用

文献类型： 中文期刊

作者： 倪健强;张春玲;童光志;仇华吉;王云峰;田志军

作者机构：

关键词： PRV;gE;鉴别诊断

期刊名称：生物工程学报

ISSN： 1000-3061

年卷期： 2004 年 20 卷 04 期

页码： 58-63

收录情况： 北大核心 ; CSCD

摘要： 根据伪狂犬病病毒 (PRV)Min A株gE基因序列 ,利用PCR方法扩增了PRVgE基因不含信号肽、胞内区和跨膜区的主要抗原表位区 ,并克隆到原核表达载体pGEX 6p 1中 ,获得的重组质粒命名为pGEX tgE。经SDS PAGE电泳分析证实克隆的部分gE基因获得了表达 ,融合表达产物大小约为 6 3kD ,并在终浓度为 0 6mmol L的IPTG诱导下 ,3 5h其表达量达到高峰。通过改变诱导条件 ,有效抑制了包涵体形成 ,提高了重组蛋白的溶解性。Westernblot分析证实表达的重组gE蛋白具有抗原反应活性。将表达产物利用亲和层析法纯化后作为ELISA抗原 ,通过对其特异性、敏感性及工作条件的优化试验 ,和对 4 8份PRV阴性血清样品的检测结果的统计学分析 ,建立了猪伪狂犬病tgE ELISA鉴别诊断方法。通过对 4 0 0份送检血清样品的检测结果分析 ,表明其与PRV全病毒ELISA试验的符合率高达 95 %以上 ,与基于抗gE蛋白单抗竞争性ELISA的符合率达 94 %。此方法可用于gE基因缺失PRV疫苗免疫动物和PRV自然感染动物的鉴别诊断

分类号： S852.65

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