【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。更多还原

【Abstract】 The main characters and antigenic epitopes of the recombinant protein Tasp-tams1-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-tams 1-spag gene and to construct prokaryotic expression vectors. The recombinant Tasp-tams1-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 11 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-tams1-spag was expressed by induction 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.更多还原