汉坦病毒囊膜糖蛋白G1基因重组腺病毒载体的构建及其在Vero E6细胞中的表达

拟构建汉坦病毒Gl基因重组腺病毒载体并在VeroE6细胞中表达，为汉坦病毒基因疫苗的研究提供实验基础。PCR法从含汉坦病毒-76118株M基因的M56质粒扩增糖蛋白G1基因片段，利用穿梭质粒pShuttle，将其克隆入Adeno—X病毒DNA，获得重组腺病毒DNA，转染HEK293细胞，包装、扩增后得到汉坦病毒Gl基因重组腺病毒原种，感染VetoE6细胞，用IFA法和ELISA法检测表达产物。得到了含汉坦病毒G1基因的重组腺病毒，其滴度约为10^11pfu／ml，感染VeroE6细胞后检测到汉坦病毒糖蛋白G1的表达。

Construction of adenoviral vector carrying Hantaan virus glycoprotein G1 gene and it's expression in Vero E6 cells

To construct a recombinant adenoviral vector carrying Hantaan virus glycoprotein G1gene and to express it in Vero E6cells.Hantaan virus glycoprotein G1gene fragment was amplified from plasmid M56by PCR,and it was cloned into Adeno-X viral DNA via pShuttle vector.The recombinant adenoviral vector was transfected into HEK293cells.Vero E6cells were infected with the recombinant adenoviral vector after it was packaged and amplified in HEK293cells.The expression of hantaan virus glycoprotein G1in infected Vero E6cells was detected by IFA and ELISA.The recombinant adenoviral vector carrying Hantaan virus glycoprotein G1gene was obtained,whose titer was10 11 pfu/ml,and Hantaan virus glycoprotein G1was detected in infected vero E6cells.The recombinant adenoviral vector carrying Hantaan virus glycoprotein G1gene is successfully constructed,paving the way for further study in Hantaan virus genic vaccine.