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目的 构建乙型脑炎病毒 (乙脑病毒 )活疫苗生产株SA1 4 14 2的感染性克隆。方法 在对SA1 4 14 2株全长基因组测序基础上 ,引入适当酶切位点和T7RNA聚合酶启动子 ,采用分部连接策略 ,获得全长基因组cDNA ,体外转录RNA后 ,转染细胞获得感染性克隆 ,病毒传代培养、抗体中和试验和乳鼠脑腔攻击试验鉴定病毒。结果 由疫苗株cDNA构建了病毒感染性克隆 ,经鉴定为乙脑病毒 ,且病毒毒力比野毒株弱。结论 获得了乙脑病毒活疫苗生产株SA1 4 14 2的感染性克隆 ,为进一步研究疫苗的减毒机理及开发新一代疫苗奠定了基础
Objective To construct an infectious clone of SA1 4 14 2 strain of Japanese encephalitis virus (JEV) live vaccine. Methods Based on the full-length genome sequencing of SA1 4 14 2, the appropriate restriction sites and T7 RNA polymerase promoter were introduced into the full-length genomic cDNA of SA1 4 14 2 to obtain the full-length genomic cDNA. After transcribing the RNA in vitro, the transfected cells were infected Viruses were identified by sex cloning, virus subculturing, antibody neutralization and neonatal rat brain challenge assays. Results The virus-infected clone was constructed from the vaccine strain cDNA, which was identified as JE virus. The virulence of the virus was weaker than that of the wild-type strain. Conclusion The infectious clone of SA1 4 14 2 strain of JE vaccine was obtained, which lays the foundation for further research on attenuated mechanism of vaccine and development of new generation vaccine