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Characterization of a novel non-muscle myosin-related protein from Onchocerca gibsoni
1Department of Molecular Sciences, James Cook University of North Queensland, Townsville, Australia.
Abstract:
A cDNA library was constructed in lambda gt11 using poly(A)+ mRNA from early larvae of Onchocerca gibsoni. Screening of the library using serum from a single onchocerciasis patient yielded several strongly immunoreactive clones, one of which (OGK2) was found to encode a novel myosin-related protein. cDNA clone OGK2 contained an insert of 2017 bp, consisting of continuous open reading frame in frame with the vector, hence this clone encodes 671 amino acid residues of a larger protein. A fragment (619 nt) of the OGK2 cDNA was subcloned into the expression vector pGEX-1N to generate a glutathione S-transferase fusion protein. Polyclonal antiserum raised to this fusion protein strongly recognised an O. gibsoni protein of approximately 220 kDa. Immunolocalization studies indicated that this protein was associated predominantly with the hypodermis and a number of other specific membrane layers in the adult parasite. Myosin-related proteins are frequently immunodominant parasite antigens and in a number of studies have been shown to confer a degree of protective immunity against the corresponding parasite. Evaluation of the protective potential of the OGK2 protein, therefore, appears to be warranted.
Insights
Researchers identified a novel myosin-related protein (OGK2) in Onchocerca gibsoni, a parasite causing onchocerciasis. This protein, found in the parasite
Area of Science:
- Parasitology
- Molecular Biology
- Immunology
Background:
- Onchocerciasis is a debilitating disease caused by the filarial nematode Onchocerca volvulus.
- Myosin-related proteins are known to be immunodominant antigens in parasitic infections.
- Identifying novel parasite antigens is crucial for developing effective diagnostics and vaccines.
Purpose of the Study:
- To identify and characterize novel antigens from Onchocerca gibsoni, a related filarial nematode.
- To investigate the potential of these antigens as targets for immune intervention against onchocerciasis.
Main Methods:
- Construction of a cDNA library from Onchocerca gibsoni early larvae.
- Screening the library with patient serum to identify immunoreactive clones.
- Characterization of the identified clone (OGK2) encoding a myosin-related protein.
- Expression and purification of a glutathione S-transferase (GST) fusion protein from OGK2 cDNA.
- Generation of polyclonal antiserum against the fusion protein.
- Immunolocalization studies to determine the protein's location within the parasite.
Main Results:
- Isolation of a cDNA clone (OGK2) encoding a novel myosin-related protein of 671 amino acids.
- Production of a GST-fusion protein from OGK2 cDNA.
- Polyclonal antiserum recognized an approximately 220 kDa Onchocerca gibsoni protein.
- Immunolocalization revealed the protein's association with the hypodermis and specific membrane layers in adult parasites.
Conclusions:
- A novel myosin-related protein (OGK2) from Onchocerca gibsoni has been identified and characterized.
- This protein is localized to the hypodermis and membranes of the adult parasite.
- The OGK2 protein represents a potential candidate antigen for evaluating protective immunity against onchocerciasis.