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Article in Korean | WPRIM | ID: wpr-127009

ABSTRACT

Japanese encephalitis virus (JEV) is a member of Flaviviruses, transmitted by mosquitoes. The core of JEV is composed of the capsid (C) proteins. In order to produce the recombinant viral C protein and the antiserum specifically recognizing the JEV C protein, we have expressed and purified the JEV C protein as a Glutathion-S-Transferase (GST) fusion protein in E. coli. The JEV C protein-coding region was PCR-amplified using the infectious cDNA of a JEV Korean isolate, and the amplicons were cloned into the pGEX4T-1 E. coli expression vector. GST-C fusion proteins were purified using a glutathione sepharose column. Subsequently, the GST-C fusion proteins were used for immunization of rabbits, and the antisera were obtained from those immunized animals. Western blot analysis using the JEV-infected BHK21 cell lysates showed that these antisera specifically reacted with the JEV C proteins. This study will provide a useful reagent for the diagnosis and understanding of the viral morphogenesis in the JEV-infected cells.


Subject(s)
Animals , Humans , Rabbits , Asian People , Blotting, Western , Capsid Proteins , Capsid , Clone Cells , Culicidae , Diagnosis , DNA, Complementary , Encephalitis Virus, Japanese , Encephalitis, Japanese , Flavivirus , Glutathione , Immune Sera , Immunization , Morphogenesis , Sepharose , Staphylococcal Protein A
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