狂犬病病毒SRV_9突变株的构建与拯救

Construction and Rescue of Rabies Virus Mutant Strain SRV_9

为构建人工修饰的狂犬病病毒,首先用人细胞色素C基因替换狂犬病病毒SRV9株基因间隔区中的非必需区域Ψ区并缺失基因组全长cDNA的糖蛋白CD编码区,得到突变型SRV9cDNA质粒。然后,该质粒与表达野生型SRV9四种结构蛋白N、P、G和L的质粒共转染BHK-21细胞。免疫荧光试验显示转染细胞中有大量特异性荧光,电子显微镜观察可见大量典型的狂犬病病毒粒子。上述结果表明已成功地拯救出了人工修饰的狂犬病病毒。狂犬病病毒SRV9突变株的成功构建与拯救,为新型狂犬病减毒活疫苗的研究提供了重要的实验工具。

To construct a rabies virus mutant,the Ψ region was replaced by the coding region of human cytochrome c gene,and the coding region for cytoplasmic domain of glycoprotein G was deleted in the full-length of genomic cDNA of rabies virus strain SRV9.The mutant plasmid and the plasmids with N,P,L and G structural proteins of wild type SRV9 were co-transfected into BHK-21 cells.It was shown by IFA that there were many specific fluorescence in the BHK-21 cells,and typical rabies virus virions were observed by electronic microscope.These results demonstrated that the mutant rabies virus was successfully rescued.The genetically modified SRV9 stain has promise to provide invaluable experimental tool to develop attenuated live rabies vaccine.