title: Construction of three new Gateway® expression plasmids for Trypanosoma cruzi.
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).
authors.affiliation: Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.
authors.forename: Victoria L
authors.initials: VL
authors.lastname: Alonso
authors.affiliation: Instituto de Biología Molecular y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas, Rosario, Argentina.
authors.forename: Carla
authors.initials: C
authors.lastname: Ritagliati
authors.affiliation: Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.
authors.forename: Pamela
authors.initials: P
authors.lastname: Cribb
authors.affiliation: Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.
authors.forename: Esteban C
authors.initials: EC
authors.lastname: Serra
mesh_terms: D015153:Blotting, Western; D020224:Expressed Sequence Tags / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins; D008018:Life Cycle Stages / Q000235:genetics; D016254:Mutagenesis, Insertional; D010957:Plasmids; D015183:Restriction Mapping / Q000379:methods*; D013752:Tetracycline / Q000494:pharmacology; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000235:genetics*
chemical_list: D049452:Green Fluorescent Proteins; D013752:Tetracycline