【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。更多还原

【Abstract】 To explore the characteristics of the Theileriaannulatasurface antigens,the surface antigen Tasp-Spag1 genes were co-expressed in prokaryotic expression system.To bioinformatically analyze the surface antigen Tasp-Spag1 genes.Tasp-Spag1 genes were amplified by PCR method to construct recombinant plasmid of pET-28a-Tasp-Spag1.After the inducible expression by IPTG,SDS-PAGE,Western blot analyses were used.Bioinformatics software was used to analyze the Tasp-Spag1 genes,the main characters and antigenic epitopes were predicted.Tasp-Spag1 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-spag1 contains 236amino acids and belongs to unstable hydrophilic nonsecreted non-transmembrane protein.It contains 33 glycosylation sites and 21 phosphoric sites,3low 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.更多还原