Journal Article

Efficient Yeast One-/Two-Hybrid Screening Using a Library Composed Only of Transcription Factors in <i>Arabidopsis thaliana</i>

Nobutaka Mitsuda, Miho Ikeda, Shinobu Takada, Yuko Takiguchi, Youichi Kondou, Takeshi Yoshizumi, Miki Fujita, Kazuo Shinozaki, Minami Matsui and Masaru Ohme-Takagi

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 51, issue 12, pages 2145-2151
Published in print December 2010 | ISSN: 0032-0781
Published online October 2010 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcq161
Efficient Yeast One-/Two-Hybrid Screening Using a Library Composed Only of Transcription Factors in Arabidopsis thaliana

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Yeast one-hybrid screening is widely used for the identification of transcription factors (TFs) that interact with specific DNA sequences. However, screening a whole cDNA library is not efficient for the identification of TFs because TF genes represent only a small percentage of clones in a cDNA library. Here, we present the development of an efficient yeast one-hybrid screening system using a prey library composed only of approximately 1,500 TF cDNAs of Arabidopsis thaliana. This library enabled us to isolate a TF that binds to a specific promoter sequence with high efficiency, even when the promoter region of the gene of interest was directly employed as bait. Furthermore, this library was also successfully applied as a yeast two-hybrid library to find TFs that interact with specific proteins. This efficient system will contribute to the elucidation of gene regulatory networks in plants.

Keywords: Arabidopsis; Methodology; Transcription factor; Yeast one-hybrid screening

Journal Article.  3659 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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