PDF(Pubmed)
Abstract：
Japanese encephalitis virus (JEV) is a pathogen responsible for high mortality and morbidity rates among children with encephalitis. Since JEV genotype 1 (GI) is the most prevalent strain in South Korea these days, corresponding research and vaccine development is urgently required. Molecular genetic studies on JEV vaccines can be boosted by obtaining genetically stable full-length infectious JEV complementary DNA (cDNA) clones. Furthermore, the significance of the reverse genetics system in facilitating molecular biological analyses of JEV properties has been demonstrated. This study constructed a recombinant JEV-GI strain using a reverse genetics system based on a Korean wild-type GI isolate (K05GS). RNA extracted from JEV-GI was used to synthesize cDNA, a recombinant full-length JEV clone, pTRE-JEVGI, was generated from the DNA fragment, and the virus was rescued. We performed in vitro and in vivo experiments to analyze the rescued JEV-GI virus. The rescued JEV-GI exhibited similar characteristics to wild-type JEV. These results suggest that our reverse genetics system can generate full-length infectious clones that can be used to analyze molecular biological factors that influence viral properties and immunogenicity. Additionally, it may be useful as a heterologous gene expression vector and help develop new strains for JEV vaccines.
摘要：
日本脑炎病毒（JEV）是导致脑炎儿童高死亡率和发病率的病原体。由于JEV基因型1（GI）是目前韩国最普遍的菌株，迫切需要相应的研究和疫苗开发。可以通过获得遗传稳定的全长感染性JEV互补DNA（cDNA）克隆来加强对JEV疫苗的分子遗传研究。此外，已经证明了反向遗传学系统在促进JEV特性的分子生物学分析中的重要性。本研究使用基于韩国野生型GI分离物（K05GS）的反向遗传系统构建了重组JEV-GI菌株。从JEV-GI中提取的RNA用于合成cDNA，重组全长JEV克隆，pTRE-JEVGI,是从DNA片段产生的，病毒获救了.我们进行了体外和体内实验以分析获救的JEV-GI病毒。获救的JEV-GI表现出与野生型JEV相似的特征。这些结果表明，我们的反向遗传学系统可以产生全长的感染性克隆，可用于分析影响病毒特性和免疫原性的分子生物学因素。此外,它可能是有用的异源基因表达载体，并有助于开发JEV疫苗的新菌株。
