Comprehensive Analysis of the DNA-Binding Specificity of an Aspergillus nidulans Transcription Factor, AmyR, Using a Bead Display System
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概要
- 論文の詳細を見る
The in vitro DNA binding profile of Aspergillus nidulans transcription factor AmyR was analyzed by a novel approach employing a genetic library of beads and flow cytometry analysis. An artificial library with 22 randomized nucleotides was constructed and subjected to a protein-DNA binding reaction with MalE-tagged AmyR. DNA fragments with potential AmyR-binding sites were labeled with fluorescence-conjugated antibody to be enriched by flow cytometry through 5 rounds of successive selection. Finally, a binding motif with a single CGG triplet was obtained from DNA fragments showing weak AmyR binding, while another motif with dual CGG triplets was discovered with stronger binding fragments. An informative motif, CGGNNNTTTNTCGG, was found to exist only in the promoter region of highly AmyR-dependent genes. These results suggest that this system is a powerful tool for the rapid and comprehensive analysis of the binding preferences of transcription factors.
- Japan Society for Bioscience, Biotechnology, and Agrochemistryの論文
- 2012-06-23
著者
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中野 秀雄
名古屋大学大学院生命農学研究科
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Kojima T
Laboratory Of Molecular Biotechnology Graduate School Of Bioagricultural Sciences Nagoya University
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Kobayashi Tetsuo
Laboratory Of Gene Regulation Graduate School Of Bioagricultural Sciences Nagoya University
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Kobayashi Tetsuo
Department Of Anatomy Ii School Of Veterinary Medicine Azabu University
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Nakano Hideo
Department Of Bioengineering Sciences Graduate School Of Bioagricultural Sciences Nagoya University
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Kojima Takaaki
Department Of Bioengineering Sciences Graduate School Of Bioagricultural Sciences Nagoya University
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WANG Panhui
Department of Bioengineering Sciences, Graduate School of Bioagricultural Sciences, Nagoya University
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Wang Panhui
Department Of Bioengineering Sciences Graduate School Of Bioagricultural Sciences Nagoya University
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KOBAYASHI Tetsuo
Department of Anatomy II
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