Toxoplasma gondii: isolation of tachyzoites rhoptries and incorporation into Iscom.
Rhoptries have been isolated from Toxoplasma gondii tachyzoites by subcellular fractionation in isopynic density sucrose gradient. Five bands were observed, and transmission electron microscopy of these indicated that rhoptries were in band 3. This band had a density of 1.17 g/cm(3). Fraction 1 had membrane structures of the parasite. Fraction 2 contained membranes and mitochondria. Fraction 4 had mostly conoid structure and fraction 5 showed ghosts. The electrophoretic and Western blotting analysis of the fractions indicated the presence of a number of proteins. Iscoms were constructed from band 3, which contained the rhoptry structures. Iscom showed a only protein incorporated of 55 kDa. Isolation of the parasite organelles has got in this work is necessary to identification, characterization, and function elucidation of the organelle proteins.
Garcia
João Luis
JL
Microbiology and Immunology Laboratory, Departamento de Biologia, Universidade Estadual do Centro Oeste-Unicentro, R. Simeão Camargo Varela de Sá, 03, Bairro Cascavel, 85040-080 Guarapuava, PR, Brazil. jlgarcia@unicentro.br
Gennari
Solange Maria
SM
Navarro
Italmar Teodorico
IT
Machado
Rosângela Zacarias
RZ
Sinhorini
Idércio Luiz
IL
D000818:Animals; D015153:Blotting, Western; D002458:Cell Fractionation; D002500:Centrifugation, Isopycnic; D004591:Electrophoresis, Polyacrylamide Gel; D016882:ISCOMs / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D015388:Organelles / Q000276:immunology / Q000502:physiology*; D016052:Protozoan Vaccines / Q000378:metabolism*; D014122:Toxoplasma / Q000276:immunology / Q000502:physiology / Q000648:ultrastructure*
D016882:ISCOMs; D016052:Protozoan Vaccines