J Bacteriol Virol.  2005 Mar;35(1):49-55.

Expression and Purification of the Capsid Protein of the Japanese Encephalitis Virus and Production of its Polyclonal Antibody

Affiliations
  • 1Department of Microbiology, College of Medicine and Medical Research Institute, Korea. ymlee@chungbuk.ac.kr
  • 2Department of Pediatrics, Clinical Research Institute, Chungbuk National University 48 Gaeshin-Dong Heungduk-Ku Cheongju Chungbuk 361-763, Korea.

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.

Keyword

Japanese encephalitis virus; Capsid protein; Protein expression and purification; Antibody

MeSH Terms

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
Capsid Proteins
DNA, Complementary
Glutathione
Immune Sera
Sepharose
Staphylococcal Protein A
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