Molecular cloning and characterization of an antigenic protein with a repeating region from Clonorchis sinensis.
10.3347/kjp.2001.39.1.57
- Author:
Tae Yun KIM
;
Shin Yong KANG
;
Il Young AHN
;
Seung Yull CHO
;
Sung Jong HONG
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- MeSH:
Amino Acid Sequence;
Animals;
Antigens, Helminth/*genetics/isolation & purification;
Base Sequence;
*Cloning, Molecular;
Clonorchis sinensis/genetics/*immunology;
DNA, Helminth;
Gene Library;
Human;
Molecular Sequence Data;
Rabbits;
Recombinant Proteins;
*Repetitive Sequences, Nucleic Acid;
Support, Non-U.S. Gov't
- From:The Korean Journal of Parasitology
2001;39(1):57-66
- CountryRepublic of Korea
- Language:English
-
Abstract:
In the course of immunoscreening of Clonorchis sinensis cDNA library, a cDNA CsRP12 containing a tandem repeat was isolated. The cDNA CsRP12 encodes two putative peptides of open reading frames (ORFs) 1 and 2 (CsRP12-1 and -2). The repetitive region is composed of 15 repeats of 10 amino acids. Of the two putative peptides, CsRP12-1 was proline-rich and found to have homologues in several organisms. Recombinant proteins of the putative peptides were bacterially produced and purified by an affinity chromatography. Recombinant CsRP12-1 protein was recognized by sera of clonorchiasis patients and experimental rabbits, but recombinant CsRP12-2 was not. One of the putative peptide, CsRP12-1, is designated CsPRA, proline-rich antigen of C. sinensis. Both the C-termini of CsRP12-1 and -2 were bacterially produced and analysed to show no antigenicity. Recombinant CsPRA protein showed high sensitivity and specificity. In experimental rabbits, IgG antibodies to CsPRA was produced between 4 and 8 weeks after the infection and decreased thereafter over one year. These results indicate that CsPRA is equivalent to a natural protein and a useful antigenic protein for serodiagnosis of human clonorchiasis.
