Construction of three new Gateway® expression plasmids for Trypanosoma cruzi
Alonso, Victoria L; Ritagliati, Carla; Cribb, Pamela; Serra, Esteban C.
Mem. Inst. Oswaldo Cruz
; 109(8): 1081-1085, 12/2014. graf
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| LILACS | ID: lil-732602
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).
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