Title DNA Sequencing and Expression of the Circumsporozoite Protein of Plasmodium vivax Korean Isolate in Escherichia coli
Author Hyeong Woo Lee 1*, Jong Soo Lee, Won Ja Lee, and Ho Sa Lee 1
Address Laboratory of Medical Zoology, Department of Viral Disease, National Institute of Health, Seoul 122-701; 1 Graduate School of Biology, Kyung Hee Unive
Bibliography Journal of Microbiology, 37(4),234-242, 1999,
DOI
Key Words Plasmodium vivax, DN sequencing, circumsporozoite protein, PV210, expression, purification
Abstract To obtain the recombinant circumsporozoite (CS) protein for the diagnosis of patients and seroepidemiology of Plasmodium vivax malaria which have been prevalent in northern part of Kyunggido, the CS protein gene was amplified by the polymerase chain reaction (PCR) from genomic DNA of the Korean vivax malaria patient. The gene consists of 1,123 nucleotides except signal peptide sequences and had an uninterrupted reading frame encoding a protein of 374 amino acids with a central region of 20 tandem repeats of the nonapeptide. The CS protein gene was expressed in Escherichia coli and purified, the molecular weight of recombinant CS protein was about 44 kDa (monomer) under denaturing purification and about 65 kDa (dimer) under native purification by SDS-PAGE. The purified recombinant CS protein which has antigenicity to malaria patients in Western blot analysis and Enzyme-linked immunosorbent assay, reacted only with the serum of P. vivax (PV210) infected malaria patients with no corss reaction to the P. falciparum malaria patient. The recombinant CS protein purified in this study will serve as a useful antigen to support the diagonis of malaria patients and seroepidemiology.
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