The functional characterisation of thousands of Trypanosoma cruzigenes remains a challenge. Reverse genetics approaches compatible with high-throughput cloningstrategies 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. cruziproteins. 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.
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
Consulta Detalhada
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