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- Title
Identification of a new gene encoding 5-enolpyruvylshikimate-3-phosphate synthase using genomic library construction strategy.
- Authors
Zhou, Chang-Yan; Tian, Yong-Sheng; Xu, Zhi-Sheng; Zhao, Wei; Chen, Chen; Bao, Wen-Hua; Bian, Lin; Cai, Run; Wu, Ai-Zhong
- Abstract
Applying the genomic library construction strategy and colony screening, a new aroA gene encoding 5-enolpyruvylshikimate-3-phosphate synthase has been identified, cloned and overexpressed in Escherichia coli, and the enzyme was purified to homogeneity. Kinetic analysis of the AroA indicated that the full-length enzyme exhibits 10-fold increased IC50 and an approximately 38-fold increased K for glyphosate compared to those of the AroA, while retaining high affinity for the substrate phosphoenolpyruvate. Furthermore, we have transformed the new aroA gene into Arabidopsis thaliana via a floral dip method, and demonstrated that transgenic A. thaliana plants exhibit significant glyphosate resistance when compared with the wild type.
- Subjects
PSEUDOMONAS fluorescens; GENE libraries; PHOSPHATES; ENZYME kinetics; PYRUVATE kinase
- Publication
Molecular Biology Reports, 2012, Vol 39, Issue 12, p10939
- ISSN
0301-4851
- Publication type
Article
- DOI
10.1007/s11033-012-1994-0