环形泰勒虫表面抗原Tams1-Spag1串联表达及其蛋白的生物信息学分析

Expression and bioinformatics analysis of Theileria annulata surface antigen Tams1-Spag1

为了研究环形泰勒虫表面抗原特性,以原核表达系统串联表达表面抗原Tams1-Spag1基因,并对其蛋白进行生物信息学分析。通过PCR技术扩增Tams1-Spag1基因片段后构建重组质粒p ET-28a-Tams1-Spag1,IPTG诱导重组蛋白表达,SDS-PAGE与Western-blot检测显示,得到大小与预期分子量相当的目的蛋白;生物信息学分析发现,此串联重组蛋白Tams1-Spag1具有219个氨基酸,属于非分泌型亲水性蛋白,分别具有13个与11个可能的糖基化与磷酸化位点,存在两个低复杂性的结构域,一个NOT的结构域,预测有6个B细胞表位优势区段与10个T细胞表位优势区段,存在两段交叉反应性表位肽。本研究为深入探讨串联重组蛋白的免疫原性提供理论依据。

To investigate the characteristics of the Theileria annulata surface antigens,the genes of the surface antigens Tams1 and Spag1 were co-expressed in bacterial expression system. The Tams1-Spag1 gene was amplified by PCR method and the recombinant plasmid p ET-28a-Tams1-Spag1 was constructed. After the recombinant protein was induced to express by IPTG,the results of SDS-PAGE and Westernblot showed that the target protein is of the expected size. Based on the bioinformatics analysis,the recombinant protein tams1-spag1 contains 219 amino acids and belongs to stable hydrophilic non-secreted protein. It contains 13 glycosylation sites,11 phosphoric sites,2 low complexity domain and NOT homologous regions. There may be 6 B-cell major epitopes,10 T-cell major epitopes and two cross-reactive epitopes. This study provides theoretical basis for investigation of the immunogenicity of the recombinant protein.