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

Construction and expression of in vitro transcription vector of HEV Ch-S-1 fulllength gene

目的:构建出HEV Ch-S-1株的全长c DNA克隆,并转染至Hep G2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p KS-HEV,成功获得HEV基因组RNA,并转染至Hep G2细胞,通过RT-n PCR、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c DNA体外转录载体p KS-HEV,并证明了该c DNA克隆在Hep G2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p KS-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。

Objective： HEV Ch-S-1 full-length c DNA clone was constructed,and transfected into Hep G2 cells to analyze its expression. Methods： HEV Ch-S-1 full length PCR product was obtained by using overlapping PCR; HEV genome RNA was obtained by constructing transcription vector p KS-HEV,and transfected into Hep G2 cells then expression of virus was detected by RT-n PCR,nested PCR and indirect immunofluorescence. Results： The transcription vector p KS-HEV was constructed,and show that the c DNA clone was successfully expressed in Hep G2 cells. Conclusion： The transcription vector p KS-HEV was successfully constructed,and made a preliminary study of expression of HEV Ch-S-1. It provides experimental basis for future research of HEV and HEV vaccine at the molecular level.