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- Title
Efficient Yeast One-/Two-Hybrid Screening Using a Library Composed Only of Transcription Factors in Arabidopsis thaliana.
- Authors
Mitsuda, Nobutaka; Ikeda, Miho; Takada, Shinobu; Takiguchi, Yuko; Kondou, Youichi; Yoshizumi, Takeshi; Fujita, Miki; Shinozaki, Kazuo; Matsui, Minami; Ohme-Takagi, Masaru
- Abstract
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.
- Subjects
TRANSCRIPTION factors; ARABIDOPSIS thaliana; NUCLEOTIDE sequence; ANTISENSE DNA; GENETIC regulation; METHODOLOGY; BIOLOGY techniques
- Publication
Plant & Cell Physiology, 2010, Vol 51, Issue 12, p2145
- ISSN
0032-0781
- Publication type
Article
- DOI
10.1093/pcp/pcq161