Author | Araújo, Patrícia Rosa de | |
Author | Burle-Caldas, Gabriela A. | |
Author | Pereira, Rosiane Aparecida da Silva | |
Author | Bartholomeu, Daniella Castanheira | |
Author | Rocha, Wanderson Duarte da | |
Author | Teixeira, Santuza Maria Ribeiro | |
Access date | 2023-07-21T16:10:18Z | |
Available date | 2023-07-21T16:10:18Z | |
Document date | 2011 | |
Citation | ARAÚJO, Patrícia Rosa de et al. Development of a dual reporter system to identify regulatory cis-acting elements in untranslated regions of Trypanosoma cruzi mRNAs. Parasitol Int., v. 60, n. 2, p. 161-169, 2011. doi: 10.1016/j.parint.2011.01.006. | en_US |
ISSN | 1383-5769 | en_US |
URI | https://www.arca.fiocruz.br/handle/icict/59713 | |
Language | eng | en_US |
Publisher | Elsevier B.V. | en_US |
Rights | restricted access | en_US |
Title | Development of a dual reporter system to identify regulatory cis-acting elements in untranslated regions of Trypanosoma cruzi mRNAs | en_US |
Type | Article | en_US |
DOI | 10.1016/j.parint.2011.01.006 | |
Abstract | In trypanosomatids, transcription is polycistronic and gene expression control occurs mainly at the post-transcriptional level. To investigate the role of sequences present in the 3?UTR of stage-specific mRNAs of Trypanosoma cruzi, we generated a new vector, named pTcDUALuc, containing the firefly and Renilla luciferase reporter genes. To test this vector, sequences derived from the 3?UTR plus intergenic regions of the alpha tubulin gene, which is up-regulated in epimastigotes, and amastin, which is up-regulated in amastigotes, were inserted downstream from the firefly reporter gene and luciferase activity was compared in transient and stable transfected parasites. As expected, increased luciferase activity was detected in epimastigotes transiently transfected with pTcDUALuc containing tubulin sequences. Using stable transfected cell lines that were allowed to differentiate into amastigotes, we observed increased luciferase activity and mRNA levels in amastigotes transfected with pTcDUALuc containing amastin sequences. We also showed that the spliced leader sequence and poly-A tail were inserted in the predicted sites of the firefly luciferase mRNA and that deletions in the alpha tubulin 3?UTR resulted in decreased luciferase expression because it affects polyadenylation. In contrast to the constructs containing 3?UTR sequences derived from tubulin and amastin genes, the presence of the 3?UTR from a trans-sialidase gene, whose expression is higher in trypomastigotes, resulted in increased luciferase activity in trypomastigotes without a corresponding increase in luciferase mRNA levels. | en_US |
Affilliation | Fundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, Brasil/Universidade Federal de Minas Gerais. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brazil | en_US |
Affilliation | Universidade Federal de Minas Gerais. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brazil | en_US |
Affilliation | Fundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, Brasil/Universidade Federal de Minas Gerais. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brazil | en_US |
Affilliation | Universidade Federal de Minas Gerais. Departamento de Parasitologia. Belo Horizonte, MG, Brazil | en_US |
Affilliation | Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brazil | en_US |
Affilliation | Universidade Federal de Minas Gerais. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brazil | en_US |
Subject | Untranslated regions | en_US |
Subject | Luciferase | en_US |
Subject | Alpha tubulin | en_US |
Subject | Amastin | en_US |
Subject | Trans-sialidase | en_US |
Subject | rypanosoma cruzi | en_US |
Embargo date | 2099-12-31 | |