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Published on: June 25, 2017
The Opisthorchis felineus paramyosin: cDNA sequence and characterization of its recombinant fragment
A V Shustov1, A T Kotelkin, A V Sorokin
1State Research Center of Virology and Biotechnology "Vector", 630559 Koltsovo, Novosibirsk Region, Russia. asuv@vector.nsc.ru
Abstract:
The cDNA sequence of Opisthorchis felineus paramyosin (PM) was determined and shown to have 66-70% homology with two schistosomes and two cestodes. Phylogenetic analysis revealed an almost equal distance between O. felineus and these distinct clades. Because of its relatively low conservation, the PM gene may be a convenient genetic marker for studying phylogenetic relationships among platyhelminthes.A 25-kDa recombinant polypeptide corresponding to the central part of the full-length PM was produced. In Western blot analysis, murine hyperimmune serum against recombinant PM (recPM) detected 100-kDa polypeptides in the O. felineusegg and somatic antigens. Interactions of recPM with polyclonal anti-parasite antibodies and anti-recPM sera in ELISA with native antigens demonstrated that recPM carries a B cell epitope identical to the O. felineusnative antigen. Our sequence and immunologic data may be helpful in developing new diagnostic tools and candidate vaccines for O. felineus infection.
Insights
The Opisthorchis felineus paramyosin gene may serve as a genetic marker for platyhelminth studies. Its recombinant form shows potential for developing diagnostic tools and vaccines against O. felineus infection.
Area of Science:
- Parasitology
- Molecular Biology
- Immunology
Background:
- Opisthorchis felineus is a significant parasite causing human disease.
- Understanding its genetic makeup and antigenic properties is crucial for control.
Purpose of the Study:
- To sequence and analyze the paramyosin (PM) gene of O. felineus.
- To produce and immunologically characterize recombinant PM (recPM).
- To explore the potential of PM as a diagnostic and vaccine candidate.
Main Methods:
- cDNA sequencing and phylogenetic analysis.
- Production of a 25-kDa recombinant polypeptide of paramyosin.
- Western blot and ELISA using murine hyperimmune serum against recPM and native O. felineus antigens.
Main Results:
- The O. felineus PM gene exhibits 66-70% homology with schistosomes and cestodes.
- Phylogenetic analysis suggests PM gene's utility as a genetic marker for platyhelminths.
- recPM contains a B cell epitope identical to the native O. felineus antigen, detected by specific antibodies.
Conclusions:
- The paramyosin gene's sequence and immunologic data offer insights into O. felineus evolution.
- recPM holds promise for the development of novel diagnostic tools and candidate vaccines for opisthorchiasis.
