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- W1567218961 abstract "Recent research approaches and methodologies allow us to develop projects forthe characterization of proteins of a given organism in scales greater than possiblesome time ago. New technologies for sequencing, cloning and protein expressionallow the generation of large amounts of new information in relatively short time.Organisms such as Trypanosoma cruzi, which have nearly half of its genome with noknown function, require studies to assign functions to proteins of unknown function.In an effort to characterize the genes of unknown function of this parasite, a group of10 genes was selected based on their differential expression duringmetacyclogenesis. The genes were annotated as 'hypothetical protein' in publicdatabases, with domains identified in the PFAM database and with orthologs in otherorganisms. The 10 selected genes were amplified and cloned in Gateway cloningplatform and the recombinant proteins were expressed in Escherichia coli. Eight ofthe 10 genes expressed insoluble proteins which were used to obtain polyclonalserum. The sera were used in Western Blot analysis to verify protein expressionthroughout the parasite metacyclogenesis as well as cellular localization by opticaland electron microscopy. To determine cellular localization, transfection experimentswere conducted in T. cruzi for the expression of proteins fused with GFP using avector compatible with the Gateway platform. Immunoprecipitation assays were alsoperformed for the 8 expressed proteins. The results obtained in this work uncoverexperimental information of genes annotated as 'hypothetical protein of unknownfunction' and assess the possibility of implementing a systematic approach with thismethodology in medium to large scale" @default.
- W1567218961 created "2016-06-24" @default.
- W1567218961 creator A5020657222 @default.
- W1567218961 date "2009-01-01" @default.
- W1567218961 modified "2023-09-27" @default.
- W1567218961 title "Caracterização de Genes de Função Desconhecida com Expressão Associada às Formas Infectivas do Trypanosoma cruzi" @default.
- W1567218961 hasPublicationYear "2009" @default.
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