Molecular cloning of ribosomal P protein in Toxoplasma gondii and the availability to detect antibody against recombinant protein in toxoplasmosis patients.
Among the panel of monoclonal antibodies (mAb) against Toxoplasma gondii, mAb of Tg621 (Tg621) clone blotted 38 kDa protein which localized in the cytoplasm of tachyzoites by immunofluorescence microscopy. The protein was not released into the parasitophorous vacuole during or after invasion. The cDNA fragment encoding the protein was obtained by screening a T. gondii cDNA expression library with Tg621. The full length cDNA sequence was completed with 5'-RACE as 1,592 bp, which contained open reading frame of 942 bp. The deduced amino acid sequence of Tg621 consisted of a polypeptide of 313 amino acids, with significant homology to ribosomal P proteins (RPP) of other organisms especially high to those of apicomplexan species. The expressed and purified TgRPP was assayed in western blot with the sera of toxoplasmosis patients and normal sera, which resulted in the 74.0% of positive reactions in toxoplasmosis patients whereas 8.3% in normal group. Therefore, the antibody formation against TgRPP in toxoplasmosis patients was regarded as specific for T. gondii infection and suggested a potential autoantibody.
Ahn
Hye-Jin
HJ
Department of Parasitology and Catholic Institute of Parasitic Diseases, Catholic University of Korea, College of Medicine, Seoul 137-701, Korea. howoo@catholic.ac.kr
Kim
Sera
S
Nam
Ho-Woo
HW
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000953:Antigens, Protozoan / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D012269:Ribosomal Proteins / Q000235:genetics* / Q000276:immunology; D016415:Sequence Alignment; D014122:Toxoplasma / Q000235:genetics*; D014123:Toxoplasmosis / Q000097:blood / Q000469:parasitology*
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D016054:DNA, Protozoan; C064601:L12E protein, Trypanosoma cruzi; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D012269:Ribosomal Proteins