J Bacteriol Virol.  2004 Mar;34(1):75-81.

Expression of the Structural Proteins of Japanese Encephalitis Virus

• Kang SY
• Yun SI
• Lim YH
• Lee YM

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.

Keyword

Japanese encephalitis virus; Structural proteins; Heterologouos gene expression vector; Flaviviruses

MeSH Terms

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
Viral Proteins