论文部分内容阅读
狂犬病毒糖蛋白基因的痘苗病毒表达质粒 pWS 4在鸡胚细胞内与野生型痘苗病毒天坛株 (痘苗病毒 )同源重组 ,经蚀斑纯化 ,获得表达狂犬病毒糖蛋白基因的重组痘苗病毒 (重组病毒 )。该重组病毒DNAdotblot显示强阳性信号 ,间接免疫荧光试验胞膜及胞浆均见到强阳性荧光反应 ,Westernblot分析仅在 6 4ku处呈现一条狂犬病毒非融合性糖蛋白带。免疫小鼠和狗 ,第 2 8d抗狂犬病毒中和抗体滴度分别达 2 4 30和 >6 96。初免后 14d用 5 0～ 10 0LD50 狂犬病毒CVS株对小鼠脑内攻击 ,保护率达 80 %以上。致病性明显减弱。从第 11代起连续传至 30代 ,病毒纯度、病毒滴度、血凝滴度、蛋白的表达、抗原活性和保护性均无差异 ,说明该重组病毒有良好的遗传稳定性。
The vaccinia virus expressing plasmid pWS 4 of rabies virus glycoprotein was homologously recombined with the wild-type vaccinia virus Tiantan strain (vaccinia virus) in chicken embryo cells and the recombinant vaccinia virus expressing the rabies virus glycoprotein gene was obtained by plaque purification virus). The recombinant adenovirus DNAototlot showed a strongly positive signal, and strong positive fluorescence was observed in the cytoplasm and cytoplasm of indirect immunofluorescence assay. Western blot analysis showed only a rabies virus non-fusion glycoprotein band at 64 ku. Immunization of mice and dogs, anti-rabies virus anti-rabies virus antibody titers reached 2 4 30 and> 6 96, respectively. 14d after initial immunization with 50 ~ 10LD50 rabies virus CVS strain in mice brain challenge, the protection rate of more than 80%. Pathogenicity was significantly reduced. There was no difference in virus purity, virus titer, hemagglutination titer, protein expression, antigen activity and protection between the 11th passage and the 30th passage, indicating that the recombinant virus has good genetic stability.