Please use this identifier to cite or link to this item: http://sgc.anlis.gob.ar/handle/123456789/2145
Title: High level of expression of the Toxoplasma gondii-recombinant Rop2 protein in Escherichia coli as a soluble form for optimal use in diagnosis
Authors: Nigro, Mónica G 
Martín, Valentina 
Kaufer, Federico 
Carral, Liliana 
Angel, Sergio O. 
Pszenny, Viviana 
Keywords: Animales;Antígenos de Protozoos;Humanos;Proteínas de la Membrana;Proteínas Protozoarias;Proteínas Recombinantes de Fusión;Solubilidad;Toxoplasma;Toxoplasmosis;Escherichia coli;Expresión Génica;Vectores Genéticos
Issue Date: Jul-2001
Publisher: Springer
Project: datasets
Journal: Molecular biotechnology 
Abstract: 
The rhoptry 2 protein (Rop2) is an interesting protein of Toxoplasma gondii that is involved in the parasite invasion of host cell, it has three T-cell epitopes and high antigenic value. However, the expression of Rop2 as a recombinant protein in Escherichia coli is not an easy task, showing low levels of expression or degradation and solubility problems. Using a recombinant Rop2(196-561) fused to 6 histidine residues, we showed high levels of expression in bacteria growing in terrific broth. rRop2(196-561) was purified mainly as a soluble product and in high concentrations (approx 1 mg/mL) under native conditions (40 mM imidazol in phosphate buffer). However, after a cycle of freezing-thawing rRop2(196-561) became insoluble. When glycerol was added to 26%, immediately after purification, the protein stayed soluble after cycles of freezing-thawing. Finally, it was demonstrated that under these conditions soluble rRop2(196-561) keeps its diagnostic value in contrast with the insoluble protein.
Description: 
Fil: Nigro, Mónica. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología. Departamento de Parasitología Sanitaria; Argentina.

Fil: Martin, Valentina. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología. Departamento de Parasitología Sanitaria; Argentina.

Fil: Kaufer, Federico. Hospital Alemán. Laboratorio de Toxoplasmosis; Argentina.

Fil: Carral, Liliana. Hospital Alemán. Laboratorio de Toxoplasmosis; Argentina.

Fil: Angel, Sergio O. Instituto Nacional de Parasitología. Departamento de Parasitología Sanitaria; Argentina.

Fil: Pszenny, Viviana. Instituto Nacional de Parasitología. Departamento de Parasitología Sanitaria; Argentina.
URI: http://sgc.anlis.gob.ar/handle/123456789/2145
ISSN: 1073-6085
DOI: 10.1385/MB:18:3:269
Rights: Open Access
Appears in Collections:Publicaciones INP

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