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 p...

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Detalles Bibliográficos
Autores principales: Feng, Shu-Ying, Ota, Kazuhisa, Ito, Takashi
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2010
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2978385/
https://www.ncbi.nlm.nih.gov/pubmed/20798175
http://dx.doi.org/10.1093/nar/gkq757
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author Feng, Shu-Ying
Ota, Kazuhisa
Ito, Takashi
author_facet Feng, Shu-Ying
Ota, Kazuhisa
Ito, Takashi
author_sort Feng, Shu-Ying
collection PubMed
description 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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spelling pubmed-29783852010-11-12 A yeast one-hybrid system to screen for methylated DNA-binding proteins Feng, Shu-Ying Ota, Kazuhisa Ito, Takashi Nucleic Acids Res Methods Online 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. Oxford University Press 2010-11 2010-08-26 /pmc/articles/PMC2978385/ /pubmed/20798175 http://dx.doi.org/10.1093/nar/gkq757 Text en © The Author(s) 2010. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Feng, Shu-Ying
Ota, Kazuhisa
Ito, Takashi
A yeast one-hybrid system to screen for methylated DNA-binding proteins
title A yeast one-hybrid system to screen for methylated DNA-binding proteins
title_full A yeast one-hybrid system to screen for methylated DNA-binding proteins
title_fullStr A yeast one-hybrid system to screen for methylated DNA-binding proteins
title_full_unstemmed A yeast one-hybrid system to screen for methylated DNA-binding proteins
title_short A yeast one-hybrid system to screen for methylated DNA-binding proteins
title_sort yeast one-hybrid system to screen for methylated dna-binding proteins
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2978385/
https://www.ncbi.nlm.nih.gov/pubmed/20798175
http://dx.doi.org/10.1093/nar/gkq757
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