【摘要】 构建了一个可以适于人用的表达狂犬病病毒糖蛋白的重组痘菌病毒疫苗株。构建的方法是：先构建一个表达β－半乳糖苷酶基因（简称ｌａｃ基因）的不经传代细胞的重组痘苗病毒，以此为亲本株，再与载体ＰＴｋ１１ｋＲＧ（狂犬病病毒糖蛋白基因插在胸苷激酶Ｔｋ区，并置于痘苗病毒１１ｋ启动子下游，Ｔｋ侧冀序列和启动子均来自痘苗病毒天坛株），在鸡胚细胞中同源重组，以蓝色空斑中挑白斑的方法筛选阳性重组病毒，用免疫荧光方法和ＡＰＡＡＰ免疫酶法证明，该病毒能较好地表达狂犬病病毒糖蛋白。动物实验表明，该病毒能较好地诱导小鼠和狗的抗狂犬病毒的中和抗体，并对狂犬病病毒ＣＶＳ株和ＳＲＶ（街毒）的攻击有保护作用。更多还原

【Abstract】 A recombinant vaccinia virus strain was constructed which might be used for the preparation of rabies vsccine for human beings.A recombinant virus expressing beta-galactosidase (which forms blue plaques in the presence of X-gal agarose overlay) was constructed,then the recombinant virus co-transfected chicken embryo fibroplast cell with the plasmid pTK11kRG (CVS rabies glycoprotein gene under the P11 promoter was inserted into the Tk locus of the Tiantan strain of vaccinia virus ). The desired recombinant has deleted the entire beta-gal gene, which could be identified from blue plaques. The immunofluorescence and immunoenzymatic test show that the rabies glycoprotein was efficiently expressed. When inoculated into mice,the recombinant could effectively elicit neutralizing antibody and protect them against CVS and SRV rabies virus challenge.更多还原