重组杆状病毒表达HEV ORF2抗原片段  

Hepatitis E virus ORF2 gene fragment expressed by recombinant baculovirus

作　　者：杜瑞娟[1] 马雁冰[1] 唐浩[1] 庄俊英[1] 乐耕耘[1] 刘勇[1] 戴长柏[1] 孙茂盛[1] 

机构地区：[1]中国医学科学院中国协和医科大学医学生物学研究所,昆明650118

出　　处：《中华微生物学和免疫学杂志》2001年第5期560-563,共4页Chinese Journal of Microbiology and Immunology

摘　　要：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 Expression of hepatitis E virus (HEV) ORF2 gene fragment by recombinant baculovirus and the study of its immunological characters. Methods Gene fragment was inserted into transfective vector pVL1393 to construct recombinant plasmid. Mediated by Lipofectin Reagent, the recombinant vector and baculovirus linearized DNA (BaculoGold TM ) co transfected insect cell Sf9 to make recombinant baculovirus. We used immunofluorescence assay (IFA) and Western blot to analyze its immunological characters, then immunized Kunming mice. Results Recombinant baculovirus was highly expressed in insect cell, the recombinant protein was proved to be HEV specific protein by IFA, Western blot and animal experiment and it could stimulate mice to produce antibody. Conclusion HEV gene could be effectively expressed by recombinant baculovirus. The expressed product presented epitopes which were recognized by HEV antibody, and thereby possessed good antigenicity.

关 键 词：肝炎病毒戊型 杆状病毒 表达 HEV ORF2基因 

分 类 号：R392[医药卫生—免疫学]