环形泰勒虫表面抗原Tasp-Spag1基因串联表达及其蛋白生物信息学分析  

Expression and Bioinformatics Analysis of Tasp-Spag1 Genes of Theileria annulata

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

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

出　　处：《动物医学进展》2015年第5期63-68,共6页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31160505)

摘　　要：研究环形泰勒虫表面抗原特性,以原核表达系统串联表达其表面抗原Tasp-Spag1,并对其蛋白进行生物信息学分析。通过PCR技术扩增Tasp-Spag1基因片段后构建重组质粒pET-28a-Tasp-Spag1,IPTG诱导重组蛋白表达,SDS-PAGE、Western blot检测;运用生物信息学软件对Tasp-Spag1基因片段进行分析,并预测其编码蛋白的主要特性与抗原表位。结果显示,扩增得到Tasp-Spag1基因长度为729bp,原核表达后通过SDS-PAGE与Western blot检测显示,得到大小与预期分子质量相当的目的蛋白;生物信息学分析发现,此串联重组蛋白Tasp-Spag1具有236个氨基酸,属于不稳定的非分泌型、非跨膜亲水蛋白,分别具有33个与21个可能的糖基化与磷酸化位点,有三段低复杂性的结构域,并且含有Sorb、NL的同源区域,可能有10个B细胞表位优势区段与7个T细胞表位优势区段,具有两段交叉反应性表位肽。To explore the characteristics of the Theileria annulata surface antigens, the surface antigen Ta sp-Spagl genes were co-expressed in prokaryotic expression system. To bioinformatically analyze the sur face antigen Tasp-Spagl genes. Tasp-Spaglgenes were amplified by PCR method to construct recombinant plasmid of pET-28a-Tasp-Spag1. After the inducible expression by IPTG, SDS-PAGE,Western blot analy ses were used. Bioinformatics software was used to analyze the Tasp-Spagl genes,the main characters and antigenic epitopes were predicted. Tasp-Spagl genes were about 729 bp amplified by PCR, then, the result of SDS-PAGE,Western blot showed that the target protein was obtained with a molecular weight same as the expected size; Based on the bioinformatics analysis, the recombinant Tasp-spaglcontains 236 amino acids and belongs to unstable hydrophilic nonsecreted non-transmembrane protein. It contains 33 glycosy lation sites and 21 phosphoric sites, 3 low complexity domain and Sorb, NL homologous regions. There may be 10 B-cell major epitope domains and 7 T-cell major epitope domains and two cross-reactive epitopes. The predicted results indicated that two different recombinant proteins have good immunogenicity in theory.

关 键 词：环形泰勒虫 串联重组蛋白 原核表达 生物信息学 

分 类 号：S852.723[农业科学—基础兽医学]