Elimination of false positives that arise in using the two-hybrid system
Bartel, P.; Chien, C.T.; Sternglanz, R.; Fields, S.
Biotechniques 14(6): 920-924
1993
ISSN/ISBN: 0736-6205 PMID: 8333960 Document Number: 412775
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.