[Construcion of a chimeric Japanese encephalits virus/dengue virus-2].
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.
Wei
Yan
Y
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
Lu
Peng
P
Yu
Jian-shi
JS
Li
Jian-dong
JD
Liu
Qin-zhi
QZ
Zhang
Quan-fu
QF
Li
Chuan
C
Miao
Fang
F
Zhang
Shuo
S
Hang
Xiao-tong
XT
Li
De-xin
DX
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