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

被引：10
|
作者：
Feng, Shu-Ying [1 ]
Ota, Kazuhisa [1 ,2 ]
Ito, Takashi [1 ,2 ]
机构：
[1] Univ Tokyo, Grad Sch Frontier Sci, Dept Computat Biol, Tokyo 1138654, Japan
[2] Univ Tokyo, Grad Sch Sci, Dept Biophys & Biochem, Tokyo 1138654, Japan
来源：
NUCLEIC ACIDS RESEARCH | 2010年 / 38卷 / 20期
关键词：
DOMAIN; MECP2; CHROMATIN; ANIMALS; PLANTS; CPG;
D O I：
10.1093/nar/gkq757
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
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页数：6
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