pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
Autor: | Marco Aurélio Krieger, Bruno Dias Ramos, Michel Batista, Fabricio K Marchini, Fernanda G. Kugeratski, Alexandre Haruo Inoue |
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Rok vydání: | 2015 |
Předmět: |
Microbiology (medical)
lcsh:Arctic medicine. Tropical medicine lcsh:RC955-962 Trypanosoma cruzi Genetic Vectors lcsh:QR1-502 cloning Gene Expression Computational biology lcsh:Microbiology Chromatography Affinity reverse genetics Plasmid Cellular localisation plasmid vectors Technical Note Cloning Molecular Gene Expressed Sequence Tags Cloning Expressed sequence tag biology biology.organism_classification Molecular biology Reverse genetics pTcGW platform p gene characterisation Plasmids Protein overexpression |
Zdroj: | Memórias do Instituto Oswaldo Cruz, Volume: 110, Issue: 5, Pages: 687-690, Published: 21 JUL 2015 Memórias do Instituto Oswaldo Cruz Memórias do Instituto Oswaldo Cruz., Vol 110, Iss 5, Pp 687-690 (2015) |
ISSN: | 1678-8060 0074-0276 |
Popis: | 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. |
Databáze: | OpenAIRE |
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