p<italic>Tc</italic>GW plasmid vectors 1.1 version: a versatile tool for <italic>Trypanosoma cruzi</italic> gene characterisation

<p>The functional characterisation of thousands of <italic>Trypanosoma cruzi</italic> 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 p<italic>Tc</italic>GW 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 p<italic>Tc</italic>GW 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 <italic>T. cruzi</italic> proteins. The 1.1 version of p<italic>Tc</italic>GW 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 <italic>T. cruzi</italic>genes and is freely available for scientific community.