An eYFP Reporter Gene for the Yeast Two-hybrid System

作者：Damon Coralie^*; Boxus Mathieu; Twizere Jean Claude; Portetelle Daniel; Vandenbol Micheline

来源：Protein Journal, 2013, 32(2): 126-130.

DOI：10.1007/s10930-013-9467-4

• Yeast two-hybrid
• Protein-protein interaction
• Reporter gene
• Yellow fluorescent protein

The yeast two-hybrid systemis 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.

• 出版日期2013-2