Optimizing the binding activity of the AP2/ERF transcription factor with the GCC box element from Brassica napus by directed evolution

文献类型： 外文期刊

第一作者： Jin, Xiao-Fen

作者： Jin, Xiao-Fen;Zhu, Bo;Peng, Ri-He;Jiang, Hai-hua;Yao, Quan-Hong;Xiong, Ai-Sheng;Jiang, Hai-hua;Chen, Jian-Min;Zhuang, Jing;Zhang, Jian

作者机构：

关键词： AP2/ERF;GCC box;Gene shuffling;Transcription factor;Yeast one-hybrid system

期刊名称：BMB REPORTS （ 影响因子：4.778； 五年影响因子：4.377 ）

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收录情况： SCI

摘要： In this study, we cloned the ERF-B3 subfamily transcription factor gene BnaERF-B3-hy15 from Brassica napus L. Huyou15. This 600 bp gene encodes a 199 amino acid classic ethylene responsive factor (ERF), which shown no binding or very weak binding GCC box-binding activity by the yeast one-hybrid assay. We used gene shuffling and the yeast one-hybrid system to obtain three mutated sequences that can bind to the GCC box. Sequence analysis indicated that two residues, Gly156 in the AP2 domain and Phe62 at the N-terminal domain were mutated to arginine and serine, respectively. Changes of Gly156 to arginine and Phe62 to serine increased the GCC-binding activity of BnaERF-B3-hy15 and the alter of Gly156 to arginine changed the AP2-domain structure of BnaERF-B3-hy15. [BMB reports 2010; 43(8): 567-572]

分类号： Q7

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