A Taenia crassiceps cDNA sequence encoding a putative immunodiagnostic antigen for bovine cysticercosis.
A cDNA expression library was constructed in lambda gt11 using poly A mRNA from the metacestode stage of Taenia crassiceps. The library was screened with rabbit antiserum to a previously defined protein fraction from Taenia hydatigena immunodiagnostic for bovine cysticercosis and with sera from cattle with experimentally induced cysticercosis. One clone (lambda TcA2) containing a 279-bp cDNA insert, reacted strongly with both antisera. A second clone (lambda TcA5.5) revealed the full-length cDNA sequence to be 361 bp. Data from Southern blots and enzymatically amplified genomic DNA segments were consistent with multiple copies or a gene family within the genome. The lambda TcA2 cDNA insert was subcloned into the plasmid pPR987 which generated a 47-kDa maltose-binding fusion protein (TcA2-MBP). Affinity-purified TcA2-MBP antigen reacted positively by ELISA with sera from cattle with experimentally induced T. saginata infections but not with sera from cattle with Fasciola hepatica or common gastrointestinal parasite infections. Rabbit polyclonal, monospecific antisera to TcA2-MBP recognized a 10-kDa protein in the cyst fluid, body wall and excretory/secretory products of the metacestode stage of T. crassiceps and immonolocalized this protein to organelles within the matrix of the cyst wall.
Zarlenga
D S
DS
U.S. Department of Agriculture, Biosystematic Parasitology Laboratory, Beltsville, MD 20705.
Rhoads
M L
ML
al-Yaman
F M
FM
D000818:Animals; D000909:Antibodies, Helminth / Q000032:analysis; D000947:Antigens, Helminth / Q000235:genetics*; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000276:immunology; D002846:Chromatography, Affinity / Q000662:veterinary; D003001:Cloning, Molecular; D003429:Cross Reactions; D003551:Cysticercosis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D018076:DNA, Complementary / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005211:Fascioliasis / Q000276:immunology / Q000662:veterinary; D015723:Gene Library; D007150:Immunohistochemistry; D007159:Immunologic Tests / Q000662:veterinary; D008969:Molecular Sequence Data; D011994:Recombinant Proteins; D013621:Taenia / Q000235:genetics / Q000276:immunology*
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018076:DNA, Complementary; D011994:Recombinant Proteins