NDLI: Toxoplasma gondii: cloning, sequencing, expression, and antigenic characterization of ROP2, GRA5 and GRA7

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Year: 2008 Volume: 7 Issue: 1

Toxoplasma gondii: cloning, sequencing, expression, and antigenic characterization of ROP2, GRA5 and GRA7

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Vidotto, O. Kano, F. Navarro, I. T. Kawasaki, P. M. Garcia, J. L. Igarashi, M. Vidotto, M. C. Tamekuni, K. Machado, R. Z.
Description	Country affiliation: Brazil Responsible library: BR26.1 Author Affiliation: Igarashi, M ( Universidade Estadual de Londrina. Centro de Ciências Agrárias. Departamento de Medicina Veterinária Preventiva. Londrina. BR); Vidotto, O ( Universidade Estadual de Londrina. Centro de Ciências Agrárias. Departamento de Medicina Veterinária Preventiva. Londrina. BR); Vidotto, M. C ( Universidade Estadual de Londrina. Centro de Ciências Agrárias. Departamento de Medicina Veterinária Preventiva. Londrina. BR); Tamekuni, K ( Universidade Estadual de Londrina. Centro de Ciências Agrárias. Departamento de Medicina Veterinária Preventiva. Londrina. BR); Navarro, I. T ( Universidade Estadual de Londrina. Centro de Ciências Agrárias. Departamento de Medicina Veterinária Preventiva. Londrina. BR); Kawasaki, P. M ( Universidade Estadual de Londrina. Centro de Ciências Agrárias. Departamento de Medicina Veterinária Preventiva. Londrina. BR); Kano, F ( Universidade Estadual de Londrina. Centro de Ciências Agrárias. Departamento de Medicina Veterinária Preventiva. Londrina. BR); Machado, R. Z ( Universidade Estadual de São Paulo Julio de Mesquita Filho. Departamento de Patologia Veterinária. Jaboticabal. BR); Garcia, J. L ( Universidade Estadual de Londrina. Centro de Ciências Agrárias. Departamento de Medicina Veterinária Preventiva. Londrina. BR)
Abstract	Toxoplasma gondii is an intracellular obligate protozoan, which infects humans and warm-blooded animals. The aim of the present study was to clone the rop2, gra5 and gra7 genes from T. gondii RH strain and to produce recombinant proteins. The rop2, gra5 and gra7 gene fragments produced by polymerase chain reaction were cloned into the pET102/D-TOPO® vector which contains thioredoxin and polyhistidine tags at the C- and N-ends, respectively, and is expressed in Escherichia coli BL21(DE-3). The expression fusion proteins were found almost entirely in the insoluble form in the cell lysate. These recombinant proteins were purified with an Ni-NTA column. Concentrations of the recombinant antigens produced in the E. coli BL21-star ranged from 300 to 500 μg/ml growth media, which was used to immunize rabbits. We observed an identity ranging from 96 to 97% when nucleotide sequences were compared to GenBank database sequences. Immunocharacterization of proteins was made by indirect immunofluorescence assay. These proteins will be used for serodiagnosis and vaccination.
Sponsorship	Fundação Araucaria (Grant 5541)
e-ISSN	16765680
Journal	Genetics and Molecular Research
Issue Number	2
Volume Number	7
Language	English
Publisher	Fundação de Pesquisas Científicas de Ribeirão Preto
Publisher Date	2008-01-01
Publisher Place	Brazil
Access Restriction	Open
Subject Keyword	Animals Antigens, Protozoan Genetics Membrane Proteins Toxoplasma Protozoan Proteins Immunology Metabolism Blotting, Western Cloning, Molecular Electrophoresis, Polyacrylamide Gel Fluorescent Antibody Technique, Indirect Oligonucleotide Array Sequence Analysis Recombinant Fusion Proteins Discipline Genetics Discipline Molecular Biology Discipline Bioinformatics
Content Type	Text
Resource Type	Article

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Toxoplasma gondii: cloning, sequencing, expression, and antigenic characterization of ROP2, GRA5 and GRA7