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pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
Kugeratski, Fernanda G; Batista, Michel; Inoue, Alexandre Haruo; Ramos, Bruno Dias; Krieger, Marco Aurelio; Marchini/, Fabricio K.
  • Kugeratski, Fernanda G; Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba. BR
  • Batista, Michel; Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba. BR
  • Inoue, Alexandre Haruo; Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba. BR
  • Ramos, Bruno Dias; Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba. BR
  • Krieger, Marco Aurelio; Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba. BR
  • Marchini/, Fabricio K; Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba. BR
Mem. Inst. Oswaldo Cruz ; 110(5): 687-690, Aug. 2015. ilus
Article in English | LILACS | ID: lil-755906
ABSTRACT

The functional characterisation of thousands of Trypanosoma cruzi genes remains a challenge. Reverse genetics approaches compatible with high-throughput cloning strategies can provide the tool needed to tackle this challenge. We previously published the pTcGW platform, composed by plasmid vectors carrying different options of N-terminal fusion tags based on Gateway® technology. Here, we present an improved 1.1 version of pTcGW vectors, which is characterised by a fully flexible structure allowing an easy customisation of each element of the vectors in a single cloning step. Additionally, both N and C-terminal fusions are available with new tag options for protein complexes purification. Three of the newly created vectors were successfully used to determine the cellular localisation of four T. cruzi proteins. The 1.1 version of pTcGW platform can be used in a variety of assays, such as protein overexpression, identification of protein-protein interaction and protein localisation. This powerful and versatile tool allows adding valuable functional information to T. cruzigenes and is freely available for scientific community.

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Full text: Available Index: LILACS (Americas) Main subject: Trypanosoma cruzi / Genetic Vectors Language: English Journal: Mem. Inst. Oswaldo Cruz Journal subject: Tropical Medicine / Parasitology Year: 2015 Type: Article Affiliation country: Brazil Institution/Affiliation country: Fundação Oswaldo Cruz/BR

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Full text: Available Index: LILACS (Americas) Main subject: Trypanosoma cruzi / Genetic Vectors Language: English Journal: Mem. Inst. Oswaldo Cruz Journal subject: Tropical Medicine / Parasitology Year: 2015 Type: Article Affiliation country: Brazil Institution/Affiliation country: Fundação Oswaldo Cruz/BR