Construction of three new Gateway® expression plasmids for Trypanosoma cruzi
Mem. Inst. Oswaldo Cruz
; 109(8): 1081-1085, 12/2014. graf
Article
em En
| LILACS
| ID: lil-732602
Biblioteca responsável:
BR1.1
ABSTRACT
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).
Palavras-chave
Texto completo:
1
Índice:
LILACS
Assunto principal:
Plasmídeos
/
Trypanosoma cruzi
/
Mapeamento por Restrição
/
Expressão Gênica
/
Vetores Genéticos
Tipo de estudo:
Prognostic_studies
Idioma:
En
Revista:
Mem. Inst. Oswaldo Cruz
Assunto da revista:
MEDICINA TROPICAL
/
PARASITOLOGIA
Ano de publicação:
2014
Tipo de documento:
Article