ABSTRACT
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.
Subject(s)
Genes, Reporter , Two-Hybrid System Techniques , Yeasts/genetics , Base Sequence , Cloning, Molecular , Flow Cytometry , Glucan 1,4-alpha-Glucosidase/analysis , Glucan 1,4-alpha-Glucosidase/genetics , Green Fluorescent Proteins , Luminescent Proteins/analysis , Luminescent Proteins/genetics , Molecular Sequence Data , Sequence AlignmentABSTRACT
El tratamiento secundario de los biosolidos se aplica para reducir la presencia de patógenos, por lo que se hipotetizó que ese tratamiento reduciría también la concentración de elementos potencialmente tóxicos, reduciendo su poder contaminante. En experimentos de campo se determinaron los niveles de cadmio, cinc, cromo, cobre, níquel, y plomo en suelo, planta y grano de maízes abonados con biosólidos que recibieron tratamiento primario y secundario
ABSTRACT
A spontaneous kanamycin-resistant Escherichia coli mutant, showing cross resitance to five other aminoglycosides and absence of the OppA protein was isolated. [3H]- dihydrostreptomycin uptake is reduced in this mutant, implying that the oligopeptide transport system in involved in accumulation of aminoglucosides, although apparently not related with aminoglycoside permeability alteration due to bacterial adaptation to osmotic changes.
