Construcion of a chimeric Japanese encephalits virus/dengue virus-2.
- Author:
Yan WEI
1
;
Peng LU
;
Jian-shi YU
;
Jian-dong LI
;
Qin-zhi LIU
;
Quan-fu ZHANG
;
Chuan LI
;
Fang MIAO
;
Shuo ZHANG
;
Xiao-tong HANG
;
De-xin LI
Author Information
1. 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
- Publication Type:Journal Article
- MeSH:
Animals;
Blotting, Western;
Cell Line;
Cricetinae;
Dengue Virus;
genetics;
Encephalitis Viruses, Japanese;
genetics;
Genetic Vectors;
genetics;
Reassortant Viruses;
genetics;
Recombination, Genetic;
genetics;
Reverse Transcriptase Polymerase Chain Reaction
- From:
Chinese Journal of Virology
2009;25(3):185-189
- CountryChina
- Language:Chinese
-
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.
