Homologous Expression of Glycosylphosphatidylinositol-Anchored Glycoproteins in Trypanosoma cruzi
| Autor: | Ivana Ducrey, Mabel Maite Lobo, María de los Milagros Camara, Camila Centeno Camean, Virginia Balouz, Gaspar E. Canepa, Ignacio Miguel Durante, Carlos A. Buscaglia, Andrea Cecilia Mesias |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Cloning chemistry.chemical_classification biology 030231 tropical medicine Transfection biology.organism_classification Cell biology 03 medical and health sciences 030104 developmental biology 0302 clinical medicine chemistry parasitic diseases Homologous chromosome Parasite hosting Trypanosoma cruzi Glycoprotein Gene Peptide sequence |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493991471 |
| Popis: | The surface coat of Trypanosoma cruzi is covered with glycosylphosphatidylinositol (GPI)-anchored glycoproteins (GAGPs) that contribute to parasite protection and to the establishment of a persistent infection in both the insect vector and the mammalian host. Multiple GAGPs that vary by amino acid sequence and/or posttranslational modifications are co-expressed on the parasite surface coat, hence curtailing structural/functional analyses on these molecules. Studies in our lab have indicated that GAGP-tagged variants expressed by transfected parasites undergo analogous posttranslational processing than endogenous ones and therefore constitute suitable tools to overcome these limitations. In this chapter, we detail the entire methodological pipeline for the efficient homologous expression of GAGPs in T. cruzi: from a simple strategy for the simultaneously cloning and tagging of the gene of interest to the biochemical validation of the parasite-expressed product. |
| Databáze: | OpenAIRE |
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