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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

作者： Jin, Xiao-Fen 1 ; Zhu, Bo 1 ; Peng, Ri-He 1 ; Jiang, Hai-hua 1 ; Chen, Jian-Min 2 ; Zhuang, Jing 3 ; Zhang, Jian 3 ; Yao, 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai 201106, Peoples R China

2.Yangzhou Univ, Coll Life Sci & Technol, Jiangsu, Peoples R China

3.Alberta Res Council, Vegreville, AB T9C 1T4, Canada

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 ）

ISSN：

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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]

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