Detecting protein-DNA interactions using a modified yeast one-hybrid system
Detecting protein-DNA interactions using a modified yeast one-hybrid system
复制标题
使用改良酵母单杂交系统检测蛋白质-DNA 相互作用
DOI：
10.1007/978-1-4939-0805-9_5
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Takashi Ito
中科院分区：
文献类型：
--
作者：
Kazuhisa Ota;Shu-Ying Feng;Takashi Ito
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.
