Construction of three new Gateway® expression plasmids for Trypanosoma cruzi

Construction of three new Gateway® expression plasmids for Trypanosoma cruzi

We present here three expression plasmids for Trypanosoma cruzi adapted to the Gateway® recombination cloning system. Two of these plasmids were designed to express trypanosomal proteins fused to a double tag for tandem affinity purification (TAPtag). The TAPtag and Gateway® casset...

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Bibliographic Details
Published in:Memórias do Instituto Oswaldo Cruz 2014-12, Vol.109 (8), p.1081-1085
Main Authors: Alonso, Victoria L, Ritagliati, Carla, Cribb, Pamela, Serra, Esteban C
Format: Article
Language:por
Subjects:
PARASITOLOGY
TROPICAL MEDICINE
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Summary:We present here three expression plasmids for Trypanosoma cruzi adapted to the Gateway® recombination cloning system. Two of these plasmids were designed to express trypanosomal proteins fused to a double tag for tandem affinity purification (TAPtag). The TAPtag and Gateway® cassette were introduced into an episomal (pTEX) and an integrative (pTREX) plasmid. Both plasmids were assayed by introducing green fluorescent protein (GFP) by recombination and the integrity of the double-tagged protein was determined by western blotting and immunofluorescence microscopy. The third Gateway adapted vector assayed was the inducible pTcINDEX. When tested with GFP, pTcINDEX-GW showed a good response to tetracycline, being less leaky than its precursor (pTcINDEX).
ISSN:1678-8060
1678-8060