环形泰勒虫表面抗原重组蛋白Tasp-Tams1-Spag1生物信息学分析与原核表达  

BIOINFORMATICS ANALYSIS PROKARYOTIC EXPRESSION OF RECOMBINANT SURFACE ANTIGEN TASP-TAMS1-SPAG1 OF THEILERIA ANNULATA

作　　者：任方[1,2] 易忠[2] 米晓云[2] 魏婕[2] 马文戈[2] 郭会玲[2] 苗书魁[2] 汪立群 王延[2] 薛英[2] 魏玉荣[2] 黄炯 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052 [2]新疆维吾尔自治区畜牧科学院兽医研究所,乌鲁木齐830000 [3]新疆维吾尔自治区动物卫生监督所,乌鲁木齐830063

出　　处：《中国动物传染病学报》2015年第2期60-67,共8页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金项目(31160505);公益性行业(农业)科研专项(201103008)

摘　　要：运用生物信息学软件对环形泰勒虫表面抗原串联基因Tasp-Tams1-Spag1进行分析,预测其编码蛋白的主要特性与抗原表位等,并对此串联基因进行克隆与原核表达。结果表明:串联重组蛋白Tasp-Tams1-Spag1属于不稳定非分泌型亲水性蛋白;编码346个氨基酸;有4个低复杂性的结构域,且含有Sorb与Btz、NL、NOT的同源区域;可能存在11个B细胞表位优势区段与17个T细胞表位优势区段,含有T、B细胞联合表位,表明Tasp-Tams1-Spag1重组蛋白可能具有良好的免疫原性。IPTG诱导重组蛋白原核表达,结果显示,该重组蛋白以包涵体形式存在;经Western blot检测,表明此串联蛋白反应原性良好,为后续深入研究免疫抗原奠定基础。The main characters and antigenic epitopes of the recombinant protein Tasp-tamsl-spag of Theileria annulata were analyzed and predicted in the present study by using the biological software and online software in order to clone the surface antigen Tasp-tamsl- spag gene and to construct prokaryotic expression vectors. The recombinant Tasp-tamsl-spag contained 346 amino acids and belonged to unstable hydrophilic non-secreted protein. It contained 4 low complexity domains as well as Sorb, Btz, NL and NOT homologous regions. There might be ll B-cell major epitope domains, 17 T-cell major epitope domains and two cross-reactive epitopes. The recombinant protein was predicted to have good immunogenicity in theory. Subsequently, the recombinant Tasp-tamsl-spag was expressed byinduction with IPTG and expression condition was optimized. The expressed protein was mainly obtained in the form of inclusion bodies. Western blot assay also revealed that the recombinant protein had good antigenicity.

关 键 词：环形泰勒虫 重组蛋白 原核表达 生物信息学 

分 类 号：S852.723[农业科学—基础兽医学]