多头带绦虫黏着蛋白TmAdh1基因的克隆及原核表达  

Cloning and prokaryotic expression of TmAdh1 gene fromTaenia multiceps

作　　者：岳龙[1] 李文卉[1] 张念章[1] 曲自刚[1] 贾万忠[1,2] 殷宏[1,2] 付宝权[1,2] 

机构地区：[1]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兽医公共卫生重点开放实验室甘肃省动物寄生虫病重点实验室,甘肃兰州730046 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《中国兽医科学》2015年第2期126-131,共6页Chinese Veterinary Science

基　　金：甘肃省创新研究群体计划项目(1210RJIA006);国家肉牛牦牛产业技术体系项目(CARS-38)

摘　　要：为研究多头带绦虫黏着蛋白TmAdh1的功能,利用PCR分别以多头带绦虫基因组DNA和cDNA为模板扩增TmAdh1的基因组序列和cDNA序列,并利用生物信息学软件对该基因进行预测分析。将TmAdh1基因片段连接至原核表达载体pGEX4T-1进行原核表达,并利用Western-blot分析该蛋白的抗原性。结果显示,TmAdh1的基因组序列长1 848bp,由2个内含子和3个外显子组成,其开放阅读框的长度为402bp,编码133个氨基酸残基。预测分析表明,TmAdh1蛋白含有1个N端信号肽序列、1个纤连蛋白三型(FnⅢ)结构域、1个糖基化位点和8个线性B细胞抗原表位。在原核系统成功表达了约40ku的重组蛋白TmAdh1,用其免疫小鼠制备超免疫血清。Western-blot试验表明,重组蛋白TmAdh1可以与免疫小鼠血清反应。以上研究结果表明,多头带绦虫TmAdh1基因是带科绦虫宿主保护性抗原的同源基因,可作为脑多头蚴病疫苗的候选分子。In order to study the function of Taenia multiceps TmAdh1 protein,the genomic DNA sequence and cDNA sequence of TmAdh1 gene were cloned by PCR and analyzed by bioinformatics software.The TmAdh1 gene fragment was ligated into pGEX4T-1 vector and the antigenicity of recombinant TmAdh1 protein was analyzed by Western-blot.In result,the TmAdh1 genomic DNA sequence was 1 848 bp in length,consisting of 2introns and 3extrons.The open reading frame of this gene is 402 bp encoding133amino acids.Prediction analysis of the TmAdh1 protein showed that it contains a N-terminal signal peptide,a fibronectin typeⅢ domain,aglycosylation site and 8linear B epitopes.The recombinant protein TmAdh1(40ku)was successfully expressed in prokaryotic system and used to immunize mice for preparation of hyperimmune serum.Western-blot analysis showed the recombinant TmAdh1 protein can react with the immune serum.This result indicated that TmAdh1 is homologous to host protective antigen genes of Taeniidae and can be used as a potential vaccine candidate molecule for cerebral coenurosis.

关 键 词：多头带绦虫 TmAdh1 纤连蛋白三型(FnⅢ)结构域 原核表达 

分 类 号：S852.734[农业科学—基础兽医学]