A simple method for developing an infectious cDNA clone of Japanese encephalitis virus

文献类型： 外文期刊

第一作者： Zheng, Hao

作者： Zheng, Hao;Zheng, Xuchen;Tong, Wu;Liu, Fei;Liang, Chao;Wang, Tao;Gao, Fei;Li, Liwei;Shan, Tongling;Li, Guoxin;Tong, Guangzhi;Zheng, Hao;Tong, Wu;Tong, Guangzhi

作者机构：

关键词： Japanese encephalitis virus;Infectious clone;NS1 ' protein

期刊名称：VIRUS GENES （ 影响因子：2.332； 五年影响因子：2.0 ）

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收录情况： SCI

摘要： Flavivirus cDNA clones frequently demonstrate genetic instability in transformed bacteria, which hampers the construction and manipulation of cDNAs for infectious flaviviruses. In this study, we developed a stable, full-length cDNA clone, pJEHEN, of a GI JEV strain HEN0701 using a medium-copy-number pBR322 vector and propagating cDNA clones at room temperature. The virus vJEHEN recovered from the infectious clone was indistinguishable from the parent virus HEN0701 with respect to plaque morphology, growth kinetics, and virulence characteristics. A T-to-A silent mutation of nucleotide 24 of the NS2a gene was introduced into the infectious cDNA clone to eliminate frameshifting. The rescued mutant virus vJETA did not express NS1' in infected cells and showed reduced growth and neurovirulence in mice. This convenient method for the construction and manipulation of infectious JEV cDNA clones may be of use in further studies to improve our understanding of the molecular mechanisms responsible for JEV replication and pathogenesis.

分类号： R37

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