Molecular cloning and characterization of a 2C-methyl-d-erythritol 2,4-cyclodiphosphate synthase gene from Cephalotaxus harringtonia
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The full-length MECPS cDNA sequence (designated as Chmecps, GenBank Accession No.: DQ415658) was isolated by rapid amplification of cDNA ends (RACE) for the first time from Cephalotaxus harringtonia. The full-length cDNA of Chmecps was 1,146 bp containing a 753 bp open reading frame (ORF) encoding a polypeptide of 250 amino acids with a calculated mass of 26.67 kDa and an isoelectric point of 9.35. Comparative and bioinformatics analyses revealed that ChMECPS showed extensive homology with MECPSs from other plant species. Phylogenetic analysis indicated ChMECPS was more ancient than other plant MECPSs. Southern hybridization analysis of the genomic DNA showed that Chmecps was a single copy gene. Tissue expression pattern analysis revealed that ChMECPS expressed strongly in root and leaf, weakly in stem.
Keywords2C-methyl-d-erythritol 2, 4-Cyclodiphosphate synthase (MECPS) Cephalotaxus harringtonia Non-mevalonate pathway (MEP) Rapid amplification of cDNA ends (RACE)
This research was financial supported by National Natural Science Foundation of China (20572130, 30600807), Shanghai Natural Science Foundation (04ZR14051), Shanghai Science and Technology Committee and Shanghai Rising-Star Program (06QB14010).
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