Elimination of false positives that arise in using the two-hybrid system

Biotechniques. 1993 Jun;14(6):920-4.


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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.
  • Technical Report

MeSH terms

  • DNA / metabolism
  • DNA-Binding Proteins / genetics*
  • False Positive Reactions
  • Fungal Proteins / genetics*
  • Humans
  • Recombinant Fusion Proteins / genetics*
  • Saccharomyces cerevisiae Proteins*
  • Trans-Activators / genetics*
  • Transcription Factors*
  • Transformation, Genetic


  • DNA-Binding Proteins
  • Fungal Proteins
  • GAL4 protein, S cerevisiae
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • Trans-Activators
  • Transcription Factors
  • DNA