【摘要】 目的克隆狂犬病毒aG株糖蛋白(Glycoprotein)基因,构建重组真核表达载体,在CHO细胞中表达aG株糖蛋白。方法采用RTPCR,从狂犬病毒aG株基因组RNA中扩增GP基因,并将该基因克隆至真核表达载体pCIdhfr上,以脂质体介导法转染CHOdhfr-细胞。用MTX筛选的抗性克隆经ELISA、免疫荧光及Westernblot检测GP蛋白的表达。结果克隆到狂犬病毒aG株糖蛋白全长基因,并在CHO细胞中进行表达。结论成功地在CHO细胞中表达了狂犬病毒aG株的糖蛋白,为进一步开发狂犬病毒基因工程疫苗打下基础。更多还原

【Abstract】 Objective To clone the gene encoding glycoprotein (GP) of rabies virus aG strain,construct a eukaryotic expression vector and express the GP in CHO cells.Methods The gene encoding GP was amplified from the genome RNA of rabies virus aG strain,cloned into eukaryotic expression vector pCI-dhfr and transfected to CHO-dhfr cells in mediation of liposome.The transfectants were selected with methotrexate (MTX),and the expressed GP protein was analyzed by ELISA,immunofluorescence method and Western blot.Results The full-length of GP gene of rabies virus aG strain was cloned and expressed in CHO cells.Conclusion The successful expression of GP of rabies virus aG strain laid a foundation of further development of recombinant rabies vaccine.更多还原