title: Identification of a new gene encoding 5-enolpyruvylshikimate-3-phosphate synthase using genomic library construction strategy.
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(                   P.fluorescens                 ) indicated that the full-length enzyme exhibits 10-fold increased IC50 and an approximately 38-fold increased K                 (                   i                 ) for glyphosate compared to those of the AroA(                   E.coli                 ), while retaining high affinity for the substrate phosphoenolpyruvate. Furthermore, we have transformed the new aroA                 (                   P.fluorescens                 ) 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.
authors.affiliation: College of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China.
authors.forename: Chang-Yan
authors.initials: CY
authors.lastname: Zhou
authors.forename: Yong-Sheng
authors.initials: YS
authors.lastname: Tian
authors.forename: Zhi-Sheng
authors.initials: ZS
authors.lastname: Xu
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
authors.forename: Chen
authors.initials: C
authors.lastname: Chen
authors.forename: Wen-Hua
authors.initials: WH
authors.lastname: Bao
authors.forename: Lin
authors.initials: L
authors.lastname: Bian
authors.forename: Run
authors.initials: R
authors.lastname: Cai
authors.forename: Ai-Zhong
authors.initials: AZ
authors.lastname: Wu
mesh_terms: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase / Q000737:chemistry / Q000235:genetics*; D000222:Adaptation, Physiological / Q000187:drug effects; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D015698:Genomic Library; D005998:Glycine / Q000031:analogs & derivatives / Q000633:toxicity; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D018517:Plant Roots / Q000033:anatomy & histology / Q000187:drug effects; D030821:Plants, Genetically Modified; D011551:Pseudomonas fluorescens / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic / Q000187:drug effects; D000097797:Glyphosate
chemical_list: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D001426:Bacterial Proteins; D005998:Glycine; D000097797:Glyphosate