Elimination of false positives that arise in using the two-hybrid system.
The two-hybrid system is a genetic method to identify proteins that interact with a specific target protein, which is expressed in yeast as a hybrid with a DNA-binding domain. Use of this method entails transforming yeast, both with this DNA-binding domain hybrid and with a library of activation domain hybrids, followed by screening for transcriptional activation of a reporter gene. In addition to proteins that interact with the target protein, a number of false positives are identified. We provide possible explanations for these false positives along with rapid assays to eliminate them.
Bartel
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Department of Microbiology, State University of New York, Stony Brook 11794.
Chien
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CT
Sternglanz
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Fields
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D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics*; D005189:False Positive Reactions; D005656:Fungal Proteins / Q000235:genetics*; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators / Q000235:genetics*; D014157:Transcription Factors; D014170:Transformation, Genetic
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D004247:DNA