细粒棘球绦虫Eg95-5蛋白生物信息学分析及鉴定  

Bioinformatics analysis and identification of Eg95-5 protein of Echinococcus granulosus

作　　者：胡尊楠 马忠臣 何金科 徐明国 陈创夫[1,3] 王震 HU Zunnan;MA Zhongchen;HE Jinke;XU Mingguo;CHEN Chuangfu;WANG Zhen(College of Animal Science and Technology, Shihezi University, Shihezi 832000, China;College of Life Sciences, Shihezi University, Shihezi 832000, China;Co-innovation Center for Zoonotic Infectious Diseases in the Western Region, Shihezi 832000, China)

机构地区：[1]石河子大学动物科技学院,新疆石河子832000 [2]石河子大学生命科学学院,新疆石河子832000 [3]西部地区高发人兽共患传染性疾病防治协同创新中心,新疆石河子832000

出　　处：《黑龙江畜牧兽医》2019年第5期26-30,176,177,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：兵团重大科技项目(2017AA003)

摘　　要：为了制备一定纯度和浓度细粒棘球绦虫(Echinococcus granulosus, Eg)Eg95-5蛋白,进一步开发新型疫苗和建立新的疾病诊断方法,试验采用生物信息学分析、pMD19-T-Eg95-5与pET30a-Eg95-5载体的构建、Eg95-5蛋白诱导表达及纯化、免疫印迹技术(Western-blot)对Eg95-5蛋白序列和反应原性进行研究。结果表明:Eg95-5蛋白无信号肽和跨膜结构,蛋白含有6个抗原决定簇和12个磷酸化位点,二级结构以无规则卷曲(39.25%)、延伸链(33.64%)和α-螺旋(21.50%)为主,并获得了Eg95-5的三级结构模型;成功克隆并构建Eg95-5重组表达载体,获得了高纯度的Eg95-5蛋白,该蛋白可与患病动物血清发生特异性反应。说明Eg95-5蛋白结构较为保守,是无信号肽和非跨膜蛋白,作为抗原可以引起良好的免疫反应。In this study, the Eg95~5 protein sequence and reactogenicity were studied with the utilization of bioinformatics software and construction of pET30a-Eg95 -5 vector, Eg95-5 protein induced expression and purification, and Western-blott in order to prepare Eg95-5 prot& n of Echinococcus granulosus (Eg) with certain purity and concentration, further develop new vaccines and establish new disease diagnosis methods. The results showed that the Eg95-5 protein had no signal peptide and transmembrane structure, the protein contained 6 antigenic determinants and 12 phosphorylation sites, and the secondary structure was randomly coiled (39.25%), extended chain ( 33. 64%) and α-Helix (21.50%) dominated and obtained the tertiary structure model of Eg95-5. Eg95~5 recombinant expression vector were successfully cloned and constructed, and high purity Eg95-5 protein were obtained, which can be correlated specific reaction with serum of sick animals. It indicates that the Eg95-5 protein structure is relatively conservative, and it is a signal-free peptide and a non-transmembrane protein. As an antigen, it can cause a good immune response.

关 键 词：棘球蚴 细粒棘球绦虫 Eg95-5 生物信息学 原核表达 WESTERN-BLOT 

分 类 号：S852.734[农业科学—基础兽医学]