用免疫金银染色法和免疫电镜观察重组痘苗病毒对狂犬病毒糖蛋白的表达

Study on the Vaccinia Recombinant Expressing Rabies Glycoprotein by Immuno-gold Silver Staining and Immuno Electronmicroscopy

用负染及超薄切片的方法电镜观察了表达狂犬病毒糖蛋白的重组痘苗病毒ＴＴＶ１１ＫＲＧ株（简称重组病毒）的纯度、形态与结构，显示重组病毒呈典型的砖形及卵圆形。用特异的抗狂犬病毒糖蛋白的单克隆抗体为一抗，胶体金标记的羊抗鼠ＩｇＧ为二抗，作免疫金银染色，在普通光学显微镜下显示该重组病毒感染的ＣＶ－１细胞膜上存在大量金银颗粒的吸附。进一步作胶体金免疫电镜，显示该感染细胞经金标后细胞膜上存在大量电子密度致密的金颗粒，从而直观地证实了该重组病毒在感染的宿主细胞膜上有大量狂犬病毒糖蛋白的表达。

Vaccinia recombinant, TTV11KRG stain, which expressed the rabies glycoprotein was observed under electron microscope for its purity, morphology and ultrastructure by negative staining and ultrasection.The purified recombinant showed typical form of brick and oval. With the monoclonal antibody of rabies glycoprotein and goat anti-mouse IgG labeled by colloidal gold, by means of Immuno-gold silver staining, the CV-1 cells infected with the vaccinia recombinant showed large quantities of black particles around the membrane under optical microscope. By immuno electronmicroscopy, it showed large quantities of gold particles in membrance of CV-1 cells infected by TTV11KRG strain. From the results obtained it is concluded that vaccinia recombinant TTV11KRG strain can lead to expression of rabies glycoprotein.