title: An eYFP reporter gene for the yeast two-hybrid system.
abstract: The yeast two-hybrid system is a powerful tool for detecting binary protein interactions, widely used in large-scale interactome mapping. We modified two yeast strains commonly used in yeast two-hybrid experiments by integrating into their genomes a new reporter gene encoding the enhanced yellow fluorescent protein eYFP. The suitability of this reporter gene for interaction screening was evaluated by fluorescence microscopy and fluorescence-activated cell sorting analysis. The gene shows good potential as a two-hybrid reporter gene for detecting strong interactions. Gal4 transcriptional activation gives rise to sufficient fluorescence for detection with a flow cytometer, but the eYFP reporter is not sensitive enough for detecting weak or moderate interactions. This study highlights the advantages of a fluorescent reporter gene in yeast two-hybrid screening.
authors: Unité de Biologie Animale et Microbienne, Gembloux Agro-Bio Tech, Université de Liège, 6 avenue du Maréchal Juin, 5030 Gembloux, Belgium. coralie.damon@hotmail.fr
authors: Coralie
authors: C
authors: Damon
authors: Mathieu
authors: M
authors: Boxus
authors: Jean-Claude
authors: JC
authors: Twizere
authors: Daniel
authors: D
authors: Portetelle
authors: Micheline
authors: M
authors: Vandenbol
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D005818:Genetic Engineering; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques / Q000295:instrumentation*
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria
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