Cloning and Recombinant Protein Expression of VP2 Gene of Human Parvovirus B19.
- Author:
Nam Yong LEE
1
;
Eui Chong KIM
Author Information
1. Department of Clinical Pathology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Parvovirus B19;
VP2;
Recombinant protein;
ELISA
- MeSH:
Antibodies;
Blotting, Western;
Clone Cells*;
Cloning, Organism*;
DNA;
Enzyme-Linked Immunosorbent Assay;
Humans*;
Immunoenzyme Techniques;
Immunoglobulin G;
Immunoglobulin M;
Parvovirus B19, Human*;
Polymerase Chain Reaction
- From:Korean Journal of Clinical Pathology
1999;19(2):208-214
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Human parvovirus B19 (B19) is associated with a variety of clinical infections. As B19 antigen is not available in large quantity, studies on cloning and recombinant protein expression have been performed. We cloned and expressed VP2 protein of B19 viruses to obtain B19 antigen in large quantity. METHODS: VP2 gene was amplified by PCR. pQE vector was constructed and transformed into E. coli for the expression of recombinant VP2 protein. Fusion protein was purified by Ni-NTA protein purification system. Finally, the purified antigen was tested for ELISA to detect IgG and IgM antibodies in patients. RESULTS: Recombinant VP2 protein, produced by transformed E. coli and expressed, was confirmed to be 58 kD protein by Western blot. Antibodies were detected in sera from all of eight patients, who had been suspicous of B19 infection clinically and confirmed to harbor B19 DNA by PCR, in assays using ELISA kit coated with recombinant VP2. CONCLUSIONS: VP2 protein, purified by recombinant protein expression in this study, can be used for enzyme immunoassay for the detection of serum antibodies and can be devoted to the future study on vaccine development.
