小亚璃眼蜱4D8基因在毕赤酵母中的表达及免疫原性分析  

Expression of 4D8 gene of Hyalomma anatolicum anatolicum in Pichia pastorisand immunogenicity analysis of the expressed product

作　　者：赵金国[1] 刘爱红[2] 殷宏[2] 罗建勋[2] 

机构地区：[1]天水市动物疫病预防控制中心,甘肃天水741000 [2]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草食动物疫病重点实验室甘肃省动物寄生虫病重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2011年第4期375-379,共5页Chinese Veterinary Science

基　　金：国家自然科技资源共享平台项目(2005DKA21100);国家高技术研究发展计划(863)项目(2006AA10A207);国家“十一五”科技支撑计划项目(2007BAD40B06)

摘　　要：为进行抗小亚璃眼蜱疫苗的研究,采用PCR方法从重组质粒pGEM-T-pHyaa4D8中克隆了pHyaa4D8分子的基因片段,构建了真核融合表达载体pPIC9K-pHyaa4D8,在毕赤酵母中进行表达,并对重组酵母菌的发酵条件进行了优化。结果表明,表达出约19ku的目的蛋白,诱导表达的最佳时间是72h。Western-blot分析结果显示,表达产物能被半饱血小亚璃眼蜱雌性成蜱唾液腺抗原免疫兔血清识别,在19ku左右出现目的条带,说明表达的目的蛋白具有反应活性。To develope the vaccines against Hyalomma anatolicum anatolicum,a 4D8 molecule extracellular domain gene fragment（pHyaa4D8） was amplified by PCR from the recombinant plasmid pGEM-T-pHyaa4D8,and cloned into pPIC9K vector to express pHyaa4D8 in Pichia pastoris.The expression conditions were optimized.In result,the recombinant strain expressed recombinant protein with a molecular weight of 19 ku and the optimal induction time was 72 h.Western-blot analysis showed that there was a specific band of 19 ku and the recombinant protein was recognized by rabbit anti-H.anatolicum anatolicum salivary gland serum.The result showed that the expressed protein possessed reactogenicity.

关 键 词：小亚璃眼蜱 4D8基因片段 真核表达 毕赤酵母 

分 类 号：S852.746[农业科学—基础兽医学]