title: pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation.
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.
authors.affiliation: Laboratório de Genômica Funcional, Instituto Carlos Chagas, Fundação Oswaldo Cruz, Curitiba, PR, BR.
authors.forename: Fernanda G
authors.initials: FG
authors.lastname: Kugeratski
authors.affiliation: Laboratório de Genômica Funcional, Instituto Carlos Chagas, Fundação Oswaldo Cruz, Curitiba, PR, BR.
authors.forename: Michel
authors.initials: M
authors.lastname: Batista
authors.affiliation: Laboratório de Genômica Funcional, Instituto Carlos Chagas, Fundação Oswaldo Cruz, Curitiba, PR, BR.
authors.forename: Alexandre Haruo
authors.initials: AH
authors.lastname: Inoue
authors.affiliation: Laboratório de Genômica Funcional, Instituto Carlos Chagas, Fundação Oswaldo Cruz, Curitiba, PR, BR.
authors.forename: Bruno Dias
authors.initials: BD
authors.lastname: Ramos
authors.affiliation: Laboratório de Genômica Funcional, Instituto Carlos Chagas, Fundação Oswaldo Cruz, Curitiba, PR, BR.
authors.forename: Marco Aurelio
authors.initials: MA
authors.lastname: Krieger
authors.affiliation: Laboratório de Genômica Funcional, Instituto Carlos Chagas, Fundação Oswaldo Cruz, Curitiba, PR, BR.
authors.forename: Fabricio K
authors.initials: FK
authors.lastname: Marchini
mesh_terms: D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D020224:Expressed Sequence Tags / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D010957:Plasmids; D014349:Trypanosoma cruzi / Q000235:genetics*