微小牛蜱4D8基因原核表达及免疫原性分析  

Prokaryotic Expression and Immunogenicity Analysis of 4D8 Gene of Boophilus microplus

作　　者：刘军龙[1] 关贵全[1] 李有全[1] 马米玲[1] 刘爱红[1] 任巧云[1] 殷宏[1] 罗建勋[1] 

机构地区：[1]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中国农业科学院兰州兽医研究所,甘肃兰州730046

出　　处：《动物医学进展》2009年第10期11-14,共4页Progress In Veterinary Medicine

基　　金：国家自然科技资源共享平台项目(2005DKA21100);国家“十一五”科技支撑计划项目(2007BAD40B06)

摘　　要：按GenBank已知微小牛蜱的4D8基因核苷酸序列设计1对表达引物。提取微小牛蜱幼蜱总RNA,反转录合成双链cDNA,用设计的表达引物扩增出4D8基因,扩增得到的核苷酸长486 bp,编码161个氨基酸,该蛋白预计分子质量为18 ku。将该基因亚克隆到原核表达载体pGEX4T-1,构建重组原核表达载体pGEX4T-1-4D8,转化BL21宿主菌,经IPTG诱导,可成功表达。重组融合蛋白分子质量为45 ku左右,与预期大小一致。Western blot显示,兔抗微小牛蜱唾液腺、肠道和卵巢抗体能够识别重组表达蛋白。One pair of primers were designed based on the 4D8 gene of Boophilus microplus published in GenBank. 4D8 gene was amplified from total RNA of larval Boophilus microplus, the sequence result showed that the 4D8 gene has 486 bp encoding 161 amino acids, the deduced molecular weight is 18 ku. The gene was subcloned into pGEX4T-1 vector and expressed in E. coli BL21 induced by IPTG. SDSPAGE analysis showed that the recombinant protein was 45 ku in molecular weight. Western-blot analysis suggested that the recombinant protein eouble be recognized by rabbit anti B. microplus gut serum, rabbit anti B. microplus salivary gland serum and rabbit anti B. microplus ovary serum.

关 键 词：微小牛蜱 4D8基因 原核表达 

分 类 号：Q786[生物学—分子生物学] S852.746[农业科学—基础兽医学]