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Position: Home > Articles > Prokaryotic Expression and Immunogenicity Analysis of 4D8 Gene of Boophilus microplus Progress in Veterinary Medicine 2009,30 (10) 11-14

微小牛蜱4D8基因原核表达及免疫原性分析

作  者:
刘军龙;关贵全;李有全;马米玲;刘爱红;任巧云;殷宏;罗建勋
单  位:
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
关键词:
微小牛蜱;4D8基因;原核表达
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。
译  名:
Prokaryotic Expression and Immunogenicity Analysis of 4D8 Gene of Boophilus microplus
作  者:
LIU Jun-long,GUAN Gui-quan,LI You-quan,MA Mi-ling,LIU Ai-hong,REN Qiao-yun,YIN Hong,LUO Jian-xun(Key Laboratory of Veterinary Parasitology of Gansu Province,Key Laboratory of Grazing Animal Diseases MOA,State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou,Gansu,730046,China)
关键词:
Boophilus microplus;4D8 gene;prokaryotic expression
摘  要:
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.SDS-PAGE analysis showed that the recombinant protein was 45 ku in molecular weight.Western-blot analysis suggested that the recombinant protein couble be recognized by rabbit anti B.microplus gut serum,rabbit anti B.microplus salivary gland serum and rabbit anti B.microplus ovary serum.

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