Expression and Purification of the Capsid Protein of the Japanese Encephalitis Virus and Production of its Polyclonal Antibody.
- Author:
Jun Sun PARK
1
;
Jeong Min KIM
;
Sang Im YUN
;
Yu Jeong CHOI
;
Byung Hak SONG
;
Sun Mi YEON
;
Seok Yong KIM
;
Youn Soo HAHN
;
Hyeon Jin PARK
;
Mi Jeong KIM
;
Young Min LEE
Author Information
1. Department of Microbiology, College of Medicine and Medical Research Institute, Korea. ymlee@chungbuk.ac.kr
- Publication Type:Original Article
- Keywords:
Japanese encephalitis virus;
Capsid protein;
Protein expression and purification;
Antibody
- MeSH:
Animals;
Asian Continental Ancestry Group*;
Blotting, Western;
Capsid Proteins*;
Capsid*;
Clone Cells;
Culicidae;
Diagnosis;
DNA, Complementary;
Encephalitis Virus, Japanese*;
Encephalitis, Japanese*;
Flavivirus;
Glutathione;
Humans;
Immune Sera;
Immunization;
Morphogenesis;
Rabbits;
Sepharose;
Staphylococcal Protein A
- From:Journal of Bacteriology and Virology
2005;35(1):49-55
- CountryRepublic of Korea
- Language:Korean
-
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.
