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目的选择及组合戊肝病毒 (HEV)抗原基因片段 ,进行表达与免疫学特性研究。方法根据已公布的 HEV序列 ,选择三段优势抗原表位 ,经 RT- PCR扩增基因片段 ,产物以单独或经甘氨酸连接肽串联组合的方式克隆到载体 pThioHis C中 ,经 DNA序列分析后 ,将构建的重组质粒转入大肠杆菌中进行表达 ,并以 Western印迹分析初步考察其免疫学特性。结果构建的六个重组质粒在大肠杆菌中获得了不同程度的表达 ,表达的目的蛋白具有与戊型肝炎患者阳性血清特异性结合的能力。结论 HEV抗原片段及其组合在大肠杆菌中得到了高效表达 ,并为优良诊断试剂及基因工程疫苗的研究提供了基础。
Objective To select and combine HEV antigen gene fragments for expression and immunological characterization. Methods According to published HEV sequences, three dominant epitopes were selected and amplified by RT-PCR. The products were cloned into vector pThioHis C either alone or in combination with glycine linker. After DNA sequence analysis, The constructed recombinant plasmid was transformed into E. coli for expression, and its immunological properties were investigated by Western blot analysis. Results The constructed six recombinant plasmids were expressed in different degrees in E. coli, and the expressed protein had the ability to specifically bind to the positive sera of hepatitis E patients. Conclusion HEV antigen fragments and their combinations have been highly expressed in E. coli and provided the basis for the study of excellent diagnostic reagents and genetically engineered vaccines.