A yeast one-hybrid system to screen for methylated DNA-binding proteins

作者：Feng Shu Ying; Ota Kazuhisa; Ito Takashi^*

来源：Nucleic Acids Research, 2010, 38(20): e189.

DOI：10.1093/nar/gkq757

We had previously exploited a method for targeted DNA methylation in budding yeast to succeed in one-hybrid detection of methylation-dependent DNA-protein interactions. Based on this finding, we developed a yeast one-hybrid system to screen cDNA libraries for clones encoding methylated DNA-binding proteins. Concurrent use of two independent bait sequences in the same cell, or dual-bait system, effectively reduced false positive clones, which were derived from methylation-insensitive sequence-specific DNA-binding proteins. We applied the dual-bait system to screen cDNA libraries and demonstrated efficient isolation of clones for methylated DNA-binding proteins. This system would serve as a unique research tool for epigenetics.

• 出版日期2010-11