戊型肝炎病毒Ch-S-1株全长体外转录载体的构建及表达初步研究

摘要

Objective:HEV Ch-S-1 full-length cDNA clone was constructed ,and transfected into HepG 2 cells to analyze its ex-pression.Methods:HEV Ch-S-1 full length PCR product was obtained by using overlapping PCR;HEV genome RNA was obtained by constructing transcription vector pKS-HEV,and transfected into HepG2 cells then expression of virus was detected by RT-nPCR,nested PCR and indirect immunofluorescence .Results:The transcription vector pKS-HEV was constructed ,and show that the cDNA clone was successfully expressed in HepG2 cells.Conclusion:The transcription vector pKS-HEV was successfully constructed ,and made a preli-minary study of expression of HEV Ch-S-1.It provides experimental basis for future research of HEV and HEV vaccine at the molecular level.%目的:构建出HEV Ch-S-1株的全长cDNA克隆,并转染至HepG2细胞并对其表达进行初步研究. 方法:利用重叠PCR方法获得HEV Ch-S-1株全长PCR产物;构建体外转录载体pKS-HEV,成功获得HEV基因组RNA,并转染至HepG2细胞,通过RT-nPCR、套式PCR和间接免疫荧光实验检测病毒的表达. 结果:成功构建了HEV基因组全长cDNA体外转录载体pKS-HEV,并证明了该cDNA克隆在HepG2细胞中成功进行了表达. 结论:成功构建了HEV Ch-S-1株全长体外转录载体pKS-HEV,并对其表达进行了初步研究,为将来在分子水平上研究HEV及研发疫苗提供了一些实验基础.