Detecting protein – DNA interactions using a modified yeast one-hybrid system

Kazuhisa Ota, Shu Ying Feng, Takashi Ito

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)


The yeast one-hybrid (Y1H) system has been among the methods of choice to detect protein–DNA interactions. However, conventional Y1H systems with a single auxotrophic reporter gene often suffer from high incidence of false positives to demonstrate a limited power in large-scale screenings. Here we describe a refined Y1H system that uses two independent bait sequences, each controlling a distinct reporter gene integrated in the host genome. With these modifications and a method of targeted DNA methylation, we succeeded in efficient isolation of clones for methylated DNA-binding proteins from mammalian cDNA libraries.

Original languageEnglish
Pages (from-to)39-50
Number of pages12
JournalMethods in Molecular Biology
Publication statusPublished - 2014

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics


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