【摘要】 目的 :获得戊型肝炎病毒 (HEV )中和表位 p166的编码基因 ,构建基于该中和表位的HEV基因免疫的真核表达载体。方法 :应用RT PCR方法从粪便标本中扩增获得HEVORF2中G64 61～G70 3 5片段 ,通过比较核酸序列同源性鉴定其基因型 ,PCR方法扩增该片段中HEV中和表位 p166的编码基因 ,经酶切后克隆于真核表达载体 pTR42 1质粒 ,并以PCR、酶切和DNA测序加以鉴定。结果 :成功克隆了 p166的编码基因 ,其基因型别为Ⅳ型 ;经酶切鉴定和DNA测序证实p166的编码基因已成功插入pTR42 1质粒 ,且核酸序列、读码框架正确。结论 :成功构建含Ⅳ型HEV中和表位编码基因的重组质粒 ,为后续基于HEV中和表位基因免疫的研究奠定基础。更多还原

【Abstract】 Objective To construct an eukaryotic expression vector used for gene immunization based on HEV neutralization epitope (p166).Methods The gene encoding HEV neutralization epitope was obtained from stool specimens by reverse transcript polymerase chain reaction (RT?PCR) and sequenced and analyzed for HEV genotype.The gene was finally cloned into expression vector-pTR421 after digested by restriction endonucleases.Results The targeted gene was successifully amplified and clustered into HEV genotype IV.The recombinant eukaryotic expression vector (pTR421 p166) based on this gene was constructed and identified as the same as designed originally by using restriction analysis and DNA sequencing.Conclusion An eukaryotic expression vector containing HEV neutralization epitope (p166) encoding gene has been successfully constructed.更多还原