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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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3-Phosphoshikimate 1-Carboxyvinyltransferase / chemistry
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3-Phosphoshikimate 1-Carboxyvinyltransferase / genetics*
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Adaptation, Physiological / drug effects
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Amino Acid Sequence
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Arabidopsis / genetics
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism
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Genes, Bacterial / genetics*
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Genetic Vectors / genetics
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Genomic Library*
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Glycine / analogs & derivatives
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Glycine / toxicity
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Glyphosate
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Kinetics
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Models, Molecular
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Molecular Sequence Data
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Phylogeny
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Plant Roots / anatomy & histology
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Plant Roots / drug effects
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Plants, Genetically Modified
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Pseudomonas fluorescens / drug effects
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Pseudomonas fluorescens / enzymology*
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Pseudomonas fluorescens / genetics*
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Pseudomonas fluorescens / growth & development
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Sequence Alignment
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Sequence Analysis, DNA
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Transformation, Genetic / drug effects
Substances
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Bacterial Proteins
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3-Phosphoshikimate 1-Carboxyvinyltransferase
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Glycine