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Mem. Inst. Oswaldo Cruz ; 109(8): 1081-1085, 12/2014. graf
Artigo em Inglês | LILACS | ID: lil-732602

RESUMO

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).


Assuntos
Expressão Gênica/genética , Vetores Genéticos/genética , Plasmídeos , Mapeamento por Restrição/métodos , Trypanosoma cruzi/genética , Western Blotting , Etiquetas de Sequências Expressas/metabolismo , Proteínas de Fluorescência Verde , Estágios do Ciclo de Vida/genética , Mutagênese Insercional , Tetraciclina/farmacologia , Trypanosoma cruzi/efeitos dos fármacos
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