title: [Construcion of a chimeric Japanese encephalits virus/dengue virus-2].
abstract: The prM/E gene of DV2 was cloned into the JEV (SA14-14-2 strain) replicon vector which had been constructed previously, and the resulting recombinant plasmid was named pPartialdeltaprM/E. The constructed chimeric clone was linearized and then was transcripted into RNA in vitro. The produced RNA was transfected into BHK-21 cells. Five to seven days later, CPE could be observed on the transfected BHK-21cells, and then the supernatant containing the chimeric virus was collected. The Supernatant was inoculated to BHK-1 cells and C6/36 cells, respectively. CPE could be observed about 4 days post the infection of C6/36cell with the chimeric virus. The results from RT-PCR, IFA, Western blot showed that the virus contained the chimeric RNA and the envelop protein of DV2. However, the chimeric virus could not be passaged in BHK-21 cell. The successful construction of the infectious clone JE/DEN-2 laid the basis for the further research of the DV vaccine.
authors.affiliation: National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, State Key Laboratory for Molecular Virology & Genetic Engineering, Beijing 100052, China. Shumman_wei@yahoo.com.cn
authors.forename: Yan
authors.initials: Y
authors.lastname: Wei
authors.forename: Peng
authors.initials: P
authors.lastname: Lu
authors.forename: Jian-shi
authors.initials: JS
authors.lastname: Yu
authors.forename: Jian-dong
authors.initials: JD
authors.lastname: Li
authors.forename: Qin-zhi
authors.initials: QZ
authors.lastname: Liu
authors.forename: Quan-fu
authors.initials: QF
authors.lastname: Zhang
authors.forename: Chuan
authors.initials: C
authors.lastname: Li
authors.forename: Fang
authors.initials: F
authors.lastname: Miao
authors.forename: Shuo
authors.initials: S
authors.lastname: Zhang
authors.forename: Xiao-tong
authors.initials: XT
authors.lastname: Hang
authors.forename: De-xin
authors.initials: DX
authors.lastname: Li
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D006224:Cricetinae; D003716:Dengue Virus / Q000235:genetics*; D018349:Encephalitis Viruses, Japanese / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D016865:Reassortant Viruses / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction