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【目的】建立狂犬病毒的反向遗传系统,为研制不含狂犬病病毒致病性的新型安全高效的狂犬疫苗提供技术依据。【方法】本研究采用反向遗传学方法和分子克隆技术,建立了狂犬病病毒Evelyn-Rokitnicki-Abelseth(ERA)疫苗株的CMV/T7、T7启动子病毒拯救系统,构建了表达N、P、L蛋白的辅助质粒。【结果】成功拯救出野生型病毒rERA-VC,在Vero细胞上的生长动力学特性与父母本ERA相同,第三代可在Vero细胞上可获得很高的生长滴度。【结论】建立了狂犬病病毒的反向遗传系统,拯救出的野生型病毒生物学特性与父母本相同。
【Objective】 To establish a reverse genetic system of rabies virus and provide a technical basis for the development of a novel safe and efficient rabies vaccine that does not contain the pathogenicity of rabies virus. 【Method】 In this study, CMV / T7 and T7 promoter rescue vectors of rabies virus Evelyn-Rokitnicki-Abelseth (ERA) vaccine strain were established by reverse genetics and molecular cloning techniques. Protein helper plasmid. 【Result】 The results showed that rERA-VC, a wild-type virus, was successfully rescued. The growth kinetics of Vero cells were the same as that of parent ERA. The third generation could obtain high growth titer on Vero cells. 【Conclusion】 The reverse genetic system of rabies virus was established, and the biological characteristics of the rescued wild type virus were the same as those of the parents.