Isolation and characterization of a 2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase gene from Taxus media |
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Authors: | H Jin Y Gong B Guo C Qiu D Liu Z Miao X Sun K Tang |
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Institution: | (1) Shanghai Key Laboratory of Agrobiotechnology, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, School of Life Science and Technology, Shanghai Jiao Tong University, Shanghai, 200030, People’s Republic of China;(2) Shenyang Medical College, Shenyang, 110034, People’s Republic of China;(3) State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Morgan-Tan International Center for Life Sciences, Fudan University, Shanghai, 200433, People’s Republic of China;(4) Faculty of Life Science and Biotechnology, Ningbo University, Ningbo, 315211, People’s Republic of China |
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Abstract: | 2C-methyl-D-erythritol 2,4-cyclodiphosphate (MEC) synthase (MECS, EC: 4.6.1.12) is the fifth enzyme of the nonmevalonate terpenoid
pathway for isopentenyl diphosphate biosynthesis and further Taxol biosynthesis. The full-length MECS cDNA sequence (GenBank
accession number DQ286391) was cloned and characterized for the first time from Taxus media, using the Rapid Amplification of cDNA Ends (RACE) technique. The full-length cDNA of Tmmecs was 1081 bp containing a 741 bp open reading frame (ORF) encoding a peptide of 247 amino acids with a calculated molecular
mass of 26.1 kDa and an isoelectric point of 8.97. Comparative and bioinformatic analyses revealed that TmMECS had extensive
homology with MECSs from other plant species. Phylogenetic analysis indicated that TmMECS was more ancient than other plant
MECSs. Southern blot analysis revealed that Tmmecs belonged to a small gene family. Tissue expression pattern analysis indicated that Tmmecs expressed constitutively in all tissues including roots, stems and leaves. The cloning and characterization of Tmmecs will be helpful to understand more about the role of MECS involved in the Taxol biosynthesis at the molecular level.
Published in Russian in Molekulyarnaya Biologiya, 2006, Vol. 40, No. 6, pp. 1013–1020.
The article was submitted by the authors in English. |
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Keywords: | Taxus media 2C-methyl-D-erythritol 2 4-cyclodiphosphate synthase (MECS) nonmevalonate pathway Taxol biosynthesis |
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