Abstract
2-C-methyl-D-erythritol 4-phosphate cytidyltransferase (MECT), the third enzyme of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway, catalyzes formation of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol from MEP. GbMECT, presumably involved in ginkgolide biosynthesis, was cloned and characterized from Ginkgo biloba embryonic roots. The protein containing the N-terminal chloroplast transit peptide consisted of 327 amino acid residues. Complementation of GbMECT with Escherichia coli NMW33, ygbP (EcMECT) knock-out mutant, rescued the mutant, confirming the function of the protein. Transcription levels of GbMECT remained generally constant in embryonic roots and leaves for 1 month. Full 88 N-terminal residues were necessary to deliver the protein into the chloroplast as shown by protein-targeting analysis with GFP as a reporter protein in Arabidopsis thaliana protoplasts.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Arabidopsis / enzymology
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Arabidopsis / genetics
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Cell Line
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Cloning, Molecular
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DNA, Complementary / genetics
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Erythritol / analogs & derivatives*
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Erythritol / chemistry
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Erythritol / metabolism
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Evolution, Molecular
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Gene Expression Regulation, Plant
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Ginkgo biloba / enzymology*
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Ginkgo biloba / genetics*
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Ginkgo biloba / growth & development
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Ginkgolides / chemistry
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Ginkgolides / metabolism
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Molecular Sequence Data
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Mutation / genetics
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Phosphotransferases / chemistry
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Phosphotransferases / genetics*
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Phosphotransferases / metabolism*
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Phylogeny
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Protoplasts / metabolism
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Sequence Alignment
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Sugar Phosphates / chemistry
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Sugar Phosphates / metabolism*
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Transcription, Genetic / genetics
Substances
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2-C-methylerythritol 4-phosphate
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DNA, Complementary
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Ginkgolides
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Sugar Phosphates
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Phosphotransferases
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Erythritol