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Molecular cloning and characterization of 5-enolpyruvylshikimate-3-phosphate synthase gene from Convolvulus arvensis L.
Authors:Zhao-Feng Huang  Chao-Xian Zhang  Hong-Juan Huang  Shou-Hui Wei  Yan Liu  Hai-Lan Cui  Jing-Chao Chen  Long Yang  Jin-Yi Chen
Affiliation:1. Institute of Plant Protection, Key Laboratory of Weed and Rodent Biology and Management, Chinese Academy of Agricultural Sciences, Beijing, 100193, China
2. Institute of Plant Protection, Heilongjiang Academy of Land Reclamation Sciences, Harbin, 150038, China
Abstract:5-Enolpyruvylshikimate-3-phosphate synthase (EPSPS), the target enzyme for glyphosate inhibition, catalyzes an essential step in the shikimate pathway for aromatic amino acid biosynthesis. The full-length cDNA of 1,751 nucleotides (CaEPSPS, Genbank accession number: EU698030) from Convolvulus arvensis was cloned and characterized. The CaEPSPS encodes a polypeptide of 520 amino acids with a calculated molecular weight of 55.5 kDa and an isoelectric point of 7.05. The results of homology analysis revealed that CaEPSPS showed highly homologous with EPSPS proteins from other plant species. Tissue expression pattern analysis indicated that CaEPSPS was constitutively expressed in stems, leaves and roots, with lower expression in roots. CaEPSPS expression level could increase significantly with glyphosate treatment, and reached its maximum at 24 h after glyphosate application. We fused CaEPSPS to the CaMV 35S promoter and introduced the chimeric gene into Arabidopsis. The resultant expression of CaEPSPS in transgenic Arabidopsis plants exhibited enhanced tolerance to glyphosate in comparison with control.
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