戊型肝炎病毒ORF3基因在杆状病毒系统中的表达与免疫学性质  

Expression of Hepatitis E Virus ORF3 Gene Fragment in Baculovirus System and Its Immunological Character

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 医学生物学研究所分子生物室,昆明650118

出　　处：《中国医学科学院学报》2001年第4期378-381,共4页Acta Academiae Medicinae Sinicae

摘　　要：目的 利用杆状病毒系统表达戊型肝炎病毒(HEV)ORF3，为进一步研究免疫学性质与功能提供基础。方法 通过RT-PCR方法获得HEV ORF3基因的目的基因片段，插入中间转染载体质粒pVL1393，构建重组质粒，再与杆状病毒线性DNA(BaculoGoldTM)共转染Sf9昆虫细胞，通过同源重组得到重组杆状病毒。以此重组杆状病毒感染昆虫细胞后，对表达的重组蛋白进行免疫学检测并免疫小鼠。结果 表达的重组蛋白约占昆虫细胞总蛋白的2%，可被戊型肝炎患者与实验猴阳性抗血清有效识别，并能诱导小鼠产生对HEV特异性免疫应答。结论 重组杆状病毒可高效表达HEV ORF3基因片段，表达产物具有HEV特异的免疫原性。Objective To express hepatitis E virus(HEV)ORF3 protein by baculovirus system and provide basis for immunological character research． Methods Hepatitis E Virus ORF3 gene fragment was obtained by RT-PCR, ligated with vector pThioHisA for sequencing and then inserted into transfective vector pVL1393 to construct recombinant plasmid． Mediated by Lipofectin Reagent, the recombinant vector and baculovirus linearized DNA(BaculoGoldT M)co-transfected insect cell Sf9 to make recombinant baculovirus． Expressed ORF3 was analyzed for its immunological charater by Western blotting, and immunized Kunming Mice． Results Recombinant ORF3 protein could be recognized by the known positive serum and promoted organism to produce HEV-specific antibody． Conclusions Recombinant baculovirus can express effectively HEV ORF3, which has HEV specific immunogenic character．

关 键 词：戊型肝炎病毒 基因表达 杆状病毒 免疫应答 

分 类 号：R373.21[医药卫生—病原生物学]