Expression and Purification of the Capsid Protein of the Japanese Encephalitis Virus and Production of its Polyclonal Antibody
Journal of Bacteriology and Virology
;
: 49-55, 2005.
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Sepharose
/
Staphylococcal Protein A
/
Blotting, Western
/
Capsid
/
Immunization
/
Clone Cells
/
DNA, Complementary
/
Encephalitis, Japanese
/
Capsid Proteins
/
Asian People
Type of study:
Diagnostic study
Limits:
Animals
/
Humans
Language:
Korean
Journal:
Journal of Bacteriology and Virology
Year:
2005
Type:
Article
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