摘要
获得汉滩病毒G2 基因 ,构建其重组腺病毒并在HEK2 93细胞中包装表达 ,为研究汉滩病毒基因疫苗提供了实验基础。设计引物采用PCR从含汉滩病毒 \|76 1 1 8株M基因的M5 6质粒扩增出糖蛋白G2 基因片段 ,并将其克隆入腺病毒载体Adeno XviralDNA ,筛选获得重组腺病毒DNA ,转染HEK2 93细胞 ,包装、扩增后得到汉滩病毒G2 基因重组腺病毒原种 ;并在感染细胞内初步表达 ,用ELISA检测表达产物。得到了含汉滩病毒G2 基因的重组腺病毒 ,其滴度约为 1 0 10 pfu/mL ,同时在感染的HEK2 93细胞中检测到汉滩病毒糖蛋白G2 的表达。含汉滩病毒糖蛋白G2 基因重组腺病毒的成功构建 。
To construct a recombinant adenovirus carrying Hantaan virus glycoprotein G 2 gene and to express it in HEK293 cells. Hantaan virus glycoprotein G 2 gene fragment was amplified from plasmid M56 by PCR, and it was cloned into Adeno X viral DNA via pShuttle vector. The recombinant adenoviral vector was transfected into HEK293 cells, and was packaged in it. The expression of Hantaan virus glycoprotein G 2 in infected HEK293 cells was detected by ELISA.The recombinant adenovirus carrying Hantaan virus glycoprotein G 2 gene was obtained, whose titer was 10 10 pfu/mL, and Hantaan virus glycoprotein G 2 was detected in infected HEK293 cells. It is concluded that the recombinant adenovirus carrying Hantaan virus glycoprotein G 2 gene is successfully constructed and expressed, which paving the way for further study in Hantaan virus genic vaccine.
出处
《科学技术与工程》
2004年第5期363-366,370,共5页
Science Technology and Engineering
基金
陕西省自然科学基金 ( 2 0 0 1SM46)资助
