Expression of the Structural Proteins of Japanese Encephalitis Virus.
- Author:
Shien Young KANG
1
;
Sang Im YUN
;
Yong Hwan LIM
;
Young Min LEE
Author Information
1. Research Institute of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University, Korea.
- Publication Type:Original Article
- Keywords:
Japanese encephalitis virus;
Structural proteins;
Heterologouos gene expression vector;
Flaviviruses
- MeSH:
Asian Continental Ancestry Group*;
Child;
Clinical Coding;
Clone Cells;
Cytoplasm;
Encephalitis Virus, Japanese*;
Encephalitis, Japanese*;
Flavivirus;
Gene Expression;
Hand;
Humans;
Intracellular Membranes;
Membranes;
Morphogenesis;
Mortality;
Plasmids;
Sindbis Virus;
Transfection;
Viral Proteins;
Virion
- From:Journal of Bacteriology and Virology
2004;34(1):75-81
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Japanese encephalitis virus (JEV) is one of the most important human pathogens, which causes the permanent neuropsychiatric sequelae and even fatal diseases with high mortality and morbidity, especially among children. In this study, we expressed the structural proteins (C, prM, and E) of JEV using a Sindbis virus-based heterologous gene expression vector, the pSinRep5. We designed two expression vectors (pSinRep5/JEV C-E and pSinRep5/JEV C-NS1), which encode the precise coding sequence of JEV C-E and JEV C-NS1 proteins, respectively. These cloned JEV structural protein genes were designed to express under the Sindbis virus subgenomic promoter. Upon the transfection and expression of the pSinRep5/JEV C-E or pSinRep5/JEV C-NS1 plasmid, the transfected cells expressed approximately 55 kDa JEV E prtiens. As designed, the JEV NS1 proteins were expressed only in the SinRep5/JEV C-NS1 RNAtransfected cells. In addition, we found in the pSinRep5/JEV C-NS1-transfected cells that the viral proteins were predominantly localized around the perinuclear membranes. On the other hand, cytoplasmic staining was mainly observed in the pSinRep5/JEV C-E RNA-transfected cells in the absence of NS1 protein. Thus, our system will provide a useful tool to dissect intracellular membrane localization signals located in the JEV structural proteins without handling the infectious JEV viral particles and to characterize viral morphogenesis of this pathogen.
