Construction of three new Gateway® expression plasmids for Trypanosoma cruzi
Mem. Inst. Oswaldo Cruz
;
109(8): 1081-1085, 12/2014. graf
Article
in English
| LILACS
| ID: lil-732602
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).
Full text:
Available
Index:
LILACS (Americas)
Main subject:
Plasmids
/
Trypanosoma cruzi
/
Restriction Mapping
/
Gene Expression
/
Genetic Vectors
Type of study:
Prognostic study
Language:
English
Journal:
Mem. Inst. Oswaldo Cruz
Journal subject:
Tropical Medicine
/
Parasitology
Year:
2014
Type:
Article
Affiliation country:
Argentina
Institution/Affiliation country:
Universidad Nacional de Rosario/AR
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