|
KMID : 0624620100430080567
|
|
BMB Reports
2010 Volume.43 No. 8 p.567 ~ p.572
|
|
|
Optimizing the binding activity of the AP2/ERF transcription factor with the GCC box element from Brassica napus by directed evolution
|
|
Jin Xiao-Fen
Zhu Bo
Peng Ri-He
Jiang Hai-Hua
Chen Jian-Min
Zhuang Jing
Zhang Jian
Yao Quan-Hong
Xiong Ai-Sheng
|
|
|
Abstract
|
|
|
|
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 GCCbinding activity of BnaERF-B3-hy15 and the alter of Gly156 to arginine changed the AP2-domain structure of BnaERF-B3- hy15.
|
|
|
KEYWORD
|
|
|
AP2/ERF, GCC box, Gene shuffling, Transcription factor, Yeast one-hybrid system
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|