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目的 用重组杆状病毒表达戊型肝炎病毒 (hepatitisEvirus,HEV)ORF2基因片段 ,并对其免疫学特性进行研究。方法 与中间转染载体pVL1 3 93相连构建重组质粒 ,在Lipofectin介导下将其与杆状病毒线性DNA(BaculoGoldTM)共转染Sf9昆虫细胞得到重组病毒 ,用免疫荧光 ,Westernblot等方法对表达产物进行免疫学特性初步研究 ,并进行动物免疫试验。结果 含ORF2结构基因片段的重组杆状病毒在Sf9昆虫细胞中获得了高效表达 ,经免疫荧光 ,Westernblot,动物免疫试验证实该重组蛋白为HEV特异性蛋白 ,可刺激机体产生相应抗体。结论 重组杆状病毒能有效表达HEV抗原基因 ,所表达的ORF2重组蛋白具有HEV抗体识别的抗原表位 ,具有较好的抗原性和免疫原性。
Objective To express the ORF2 gene fragment of hepatitis E virus (HEV) by recombinant baculovirus and study its immunological properties. Methods Recombinant plasmids were constructed by ligating with the intermediate transfection vector pVL1 3 93. The recombinant plasmids were co-transfected into baculovirus Sf9 insect cells with BaculoGoldTM under Lipofectin. The recombinant viruses were obtained by immunofluorescence and Western blot. Immunological characteristics of the initial study, and animal immunization test. Results The recombinant baculovirus containing ORF2 gene fragment was highly expressed in Sf9 insect cells. The recombinant protein was identified as HEV by immunofluorescence, Western blot and animal immunoassay, which could stimulate the body to produce the corresponding antibody. Conclusion Recombinant baculovirus can effectively express HEV antigen gene. The expressed ORF2 recombinant protein has the antigen epitope recognized by HEV antibody and has good antigenicity and immunogenicity.