A yeast one-hybrid system to screen for methylated DNA-binding proteins.
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.
Feng
Shu-Ying
SY
Department of Computational Biology, Graduate School of Frontier Sciences, University of Tokyo, Tokyo, Japan.
Ota
Kazuhisa
K
Ito
Takashi
T
D000818:Animals; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000378:metabolism; D017930:Genes, Reporter; D006801:Humans; D051379:Mice; D012441:Saccharomyces cerevisiae / Q000235:genetics; D020798:Two-Hybrid System Techniques
D004268:DNA-Binding Proteins
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