Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 39-50 |
| Number of pages | 12 |
| Journal | Methods in Molecular Biology |
| Volume | 1164 |
| DOIs | |
| Publication status | Published - Jan 1 2014 |
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All Science Journal Classification (ASJC) codes
- Molecular Biology
- Genetics
Cite this
Detecting protein – DNA interactions using a modified yeast one-hybrid system. / Ota, Kazuhisa; Feng, Shu Ying; Ito, Takashi.
In: Methods in Molecular Biology, Vol. 1164, 01.01.2014, p. 39-50.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Detecting protein – DNA interactions using a modified yeast one-hybrid system
AU - Ota, Kazuhisa
AU - Feng, Shu Ying
AU - Ito, Takashi
PY - 2014/1/1
Y1 - 2014/1/1
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=84922480754&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-0805-9_5
DO - 10.1007/978-1-4939-0805-9_5
M3 - Article
C2 - 24927834
AN - SCOPUS:84922480754
VL - 1164
SP - 39
EP - 50
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -