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Stunnenberg, H.G., International Human Epigenome Consortium (including Ito, T.) & Hirst, M., The International Human Epigenome Consortium: A blueprint for scientific collaboration and discovery., Cell, 10.1016/j.cell.2016.11.007., 2016.11. |
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Kazuhisa Ota, Shu-Ying Feng, Takashi Ito, Detecting protein-DNA interactions using a modified yeast one-hybrid system. , Methods Mol Biol. 1164:39-50., 2014.05, 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.. |