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Genet. mol. res. (Online) ; Genet. mol. res. (Online);5(3): 553-563, 2006. ilus, graf
Artículo en Inglés | LILACS | ID: lil-441051

RESUMEN

In a 17-kb genomic fragment of Trypanosoma cruzi chromosome XX, we identified three tandemly linked genes coding for CX2CX4HX4C zinc finger proteins. We also showed that similar genes are present in T. brucei and Leishmania major, sharing three monophyletic groups among these trypanosomatids. In T. cruzi, TcZFP8 corresponds to a novel gene coding for a protein containing eight zinc finger motifs. Molecular cloning of this gene and heterologous expression as a fusion with a His-tag were performed in Escherichia coli. The purified recombinant protein was used to produce antibody in rabbits. Using Western blot analysis, we observed the presence of this protein in all three forms of the parasite: amastigote, trypomastigote and epimastigote. An analysis of cytoplasmic and nuclear cell extracts showed that this protein is present in nuclear extracts, and indirect immunofluorescence microscopy confirmed the nuclear localization of TcZFP8. Homologues of TcZFP8 in T. brucei are apparently absent, while one candidate in L. major was identified.


Asunto(s)
Animales , Conejos , Núcleo Celular/metabolismo , Código Genético/genética , Proteínas Protozoarias/genética , Trypanosoma cruzi/genética , Dedos de Zinc/genética , Secuencia de Bases , Western Blotting , Clonación Molecular , ADN Protozoario/genética , ADN Protozoario/metabolismo , Técnica del Anticuerpo Fluorescente Indirecta , Microscopía Fluorescente , Datos de Secuencia Molecular , Proteínas Protozoarias/metabolismo , ARN Protozoario/genética , ARN Protozoario/metabolismo , Trypanosoma cruzi/metabolismo
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