A 14-3-3 protein homologue is expressed in feline enteroepithelial-stages of Toxoplasma gondii.
Fourteen cDNA clones encoding epitopes of proteins of Toxoplasma gondii feline enteroepithelial-stages parasites were isolated and expressed in Escherichia coli in an effort to determine the antigenecity of the parasites. Sequence analysis showed that four of the cDNA clones had a 930-bp open-reading frame encoding a product showing similarity to the 14-3-3 protein mRNA sequence.(1) Southern hybridization of DIG-labeled positive clone with T. gondii genomic DNA cleaved with EcoRI, BamHI and HindIII resulted in one or two bands in each case. In an immunofluorescence assay, polyclonal and monoclonal antibodies raised against the expressed protein showed strong reactivity with feline enteroepithelial-stages parasites and sporozoites. In a complementation assay in which a plasmid carrying the protein-coding region of the isolated cDNA was introduced into a Saccharomyces cerevisiae mutant, strain DS9-22, the expressed protein showed complementation of the function of the 14-3-3 protein in yeast transformants. These findings suggest that T. gondii parasites produce a protein showing partial homology with members of the 14-3-3 protein family and this protein is expressed in feline enteroepithelial-stages parasites.
Koyama
T
T
Department of Basic Veterinary Science, The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan.
Ohsawa
T
T
Shimada
S
S
Omata
Y
Y
Xuan
X
X
Inoue
N
N
Maeda
R
R
Mikami
T
T
Saito
A
A
D048948:14-3-3 Proteins; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D015139:Blotting, Southern / Q000662:veterinary; D002371:Cat Diseases / Q000469:parasitology*; D002415:Cats / Q000469:parasitology; D018076:DNA, Complementary / Q000235:genetics; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D015723:Gene Library; D005816:Genetic Complementation Test / Q000662:veterinary; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013047:Specific Pathogen-Free Organisms; D014122:Toxoplasma / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014124:Toxoplasmosis, Animal / Q000097:blood / Q000276:immunology / Q000469:parasitology*; D014446:Tyrosine 3-Monooxygenase / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
D048948:14-3-3 Proteins; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D018076:DNA, Complementary; D000939:Epitopes; D014446:Tyrosine 3-Monooxygenase