Production of a recombinant fusion protein of Sarcocystis tenella and evaluation of its diagnostic potential in an ELISA

We have isolated a cDNA clone encoding an antigenic polypeptide of Sarcocystis tenella by screening a cystozoite-derived cDNA library λgt11 with antibodies from sheep infected experimentally with S. tenella. This clone, termed STC29, was subcloned and expressed in the vector pGEX-3X as a soluble fus...

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Bibliographic Details
Published in:Veterinary parasitology 1996-10, Vol.65 (3), p.185-197
Main Authors: Mertens, C.M., Tenter, A.M., Vietmeyer, C., Ellis, J.T., Johnson, A.M.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibodies, Protozoan - blood
Antibodies, Protozoan - immunology
Antigens, Protozoan - genetics
Antigens, Protozoan - immunology
Base Sequence
Blotting, Western - veterinary
Cloning, Molecular
DNA, Protozoan - chemistry
ELISA
Enzyme-Linked Immunosorbent Assay - veterinary
Female
Gene Expression
GST
Immune Sera - immunology
Mice
Molecular Sequence Data
Polymerase Chain Reaction - veterinary
Recombinant antigen
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - immunology
Sarcocystis - genetics
Sarcocystis - immunology
Sarcocystis tenella
Sarcocystosis - diagnosis
Sarcocystosis - immunology
Sarcocystosis - veterinary
Sequence Alignment - veterinary
Sheep
Sheep Diseases - diagnosis
Sheep Diseases - immunology
Sheep-Protozoa
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Summary:We have isolated a cDNA clone encoding an antigenic polypeptide of Sarcocystis tenella by screening a cystozoite-derived cDNA library λgt11 with antibodies from sheep infected experimentally with S. tenella. This clone, termed STC29, was subcloned and expressed in the vector pGEX-3X as a soluble fusion protein with glutathione S-transferase having an apparent molecular mass of 46 kDa. Antibody raised against the recombinant fusion protein recognized a native polypeptide of 25 kDa in cystozoites of S. tenella. In an ELISA with sera from experimentally infected sheep, the recombinant STC29 antigen could differentiate infections with S. tenella from those with Sarcocystis arieticanis or Toxoplasma gondii. Hence, the research described here reports the identification of the first recombinant S. tenella antigen that may be useful for standardization of a serological test for the diagnosis of S. tenella infections in sheep.
ISSN:0304-4017
1873-2550
DOI:10.1016/S0304-4017(96)00971-5