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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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