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拟构建汉坦病毒G1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G1基因重组腺病毒原种,感染VeroE6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1011pfu/ml,感染VeroE6细胞后检测到汉坦病毒糖蛋白G1的表达。
The recombinant adenovirus vector of Hantavirus G1 gene is to be constructed and expressed in VeroE6 cells, which provides experimental basis for the study of Hantavirus gene vaccine. PCR was used to amplify glycoprotein G1 gene fragment from M56 plasmid containing Hantavirus-76118 M gene. The recombinant plasmid was cloned into Adeno-X virus DNA using shuttle plasmid pShuttle to obtain recombinant adenovirus DNA, which was transfected into HEK293 cells and packaged After amplification, the recombinant adenovirus of Hantaan virus G1 gene was obtained and infected VeroE6 cells. The expression products were detected by IFA and ELISA. The recombinant adenovirus containing the Hantaan virus G1 gene was obtained with a titer of about 1011 pfu / ml. The expression of Hantaan virus glycoprotein G1 was detected after infection with VeroE6 cells.