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

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• 作者：Hao Zheng ; Xuchen Zheng ; Wu Tong ; Fei Liu ; Chao Liang ; Tao Wang ; Fei Gao…
• 关键词：Japanese encephalitis virus ; Infectious clone ; NS1′ protein
• 刊名：Virus Genes
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：53
• 期：1
• 页码：4-14
• 全文大小：
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Medical Microbiology; Virology; Plant Sciences;
• 出版者：Springer US
• ISSN：1572-994X
• 卷排序：53

文摘

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.