High level of expression of the Toxoplasma gondii-recombinant Rop2 protein in Escherichia coli as a soluble form for optimal use in diagnosis

dc.creatorNigro, Mónica G
dc.creatorMartín, Valentina
dc.creatorKaufer, Federico
dc.creatorCarral, Liliana
dc.creatorAngel, Sergio O.
dc.creatorPszenny, Viviana
dc.date2021-01-15T16:06:00Z
dc.date2021-01-15T16:06:00Z
dc.date2001-07
dc.date.accessioned2023-08-29T20:08:49Z
dc.date.available2023-08-29T20:08:49Z
dc.identifier1073-6085
dc.identifierhttp://sgc.anlis.gob.ar/handle/123456789/2145
dc.identifier10.1385/MB:18:3:269
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8520360
dc.descriptionFil: Nigro, Mónica. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología. Departamento de Parasitología Sanitaria; Argentina.
dc.descriptionFil: Martin, Valentina. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología. Departamento de Parasitología Sanitaria; Argentina.
dc.descriptionFil: Kaufer, Federico. Hospital Alemán. Laboratorio de Toxoplasmosis; Argentina.
dc.descriptionFil: Carral, Liliana. Hospital Alemán. Laboratorio de Toxoplasmosis; Argentina.
dc.descriptionFil: Angel, Sergio O. Instituto Nacional de Parasitología. Departamento de Parasitología Sanitaria; Argentina.
dc.descriptionFil: Pszenny, Viviana. Instituto Nacional de Parasitología. Departamento de Parasitología Sanitaria; Argentina.
dc.descriptionThe 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.
dc.languageen
dc.publisherSpringer
dc.relation#PLACEHOLDER_PARENT_METADATA_VALUE#
dc.relationdatasets
dc.relationMolecular biotechnology
dc.rightsnone
dc.sourceMolecular Biotechnology 2001; 18(3):269-273
dc.subjectAnimales
dc.subjectAntígenos de Protozoos
dc.subjectHumanos
dc.subjectProteínas de la Membrana
dc.subjectProteínas Protozoarias
dc.subjectProteínas Recombinantes de Fusión
dc.subjectSolubilidad
dc.subjectToxoplasma
dc.subjectToxoplasmosis
dc.subjectEscherichia coli
dc.subjectExpresión Génica
dc.subjectVectores Genéticos
dc.titleHigh level of expression of the Toxoplasma gondii-recombinant Rop2 protein in Escherichia coli as a soluble form for optimal use in diagnosis
dc.typeArtículo


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