Evaluation of alternative reporter genes for the yeast two-hybrid system
Genet. mol. res. (Online)
; 2(1): 124-135, Mar. 2003.
Article
in En
| LILACS
| ID: lil-417617
Responsible library:
BR1.1
RESUMO
The yeast two-hybrid system is a powerful tool for screening protein-protein interactions and has also been used for large-scale studies. We evaluated two protein-coding sequences as reporter genes for the yeast two-hybrid system, to determine if it was suitable as an alternative screening strategy. Aspergillus awamori glucoamylase activity results in clear haloes around colonies producing this enzyme after growth on starch plates and staining with iodine vapors. However, transcription activation by Gal4 on Gal-regulated promoters was insufficient for this type of phenotypic visualization. A modified gene of Aequoria victoria enhanced green fluorescent protein (EGFP) was tested to determine its suitability for interaction screenings with flow cytometry. When the EGFP reporter gene system was incorporated into the cells, Gal4 transcriptional activation produced sufficient fluorescence for detection with the flow cytometer, especially when there were strong interactions
Full text:
1
Collection:
01-internacional
Database:
LILACS
Main subject:
Yeasts
/
Genes, Reporter
/
Two-Hybrid System Techniques
Language:
En
Journal:
Genet. mol. res. (Online)
Journal subject:
BIOLOGIA MOLECULAR
/
GENETICA
Year:
2003
Document type:
Article
Affiliation country:
Brazil
Country of publication:
BR
/
BRASIL
/
BRASILE
/
BRAZIL
/
BRESIL