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Title:          Elucidation of the biosynthesis of carnosic acid and its reconstitution in yeast
Subject:        Nature Communications 7,  (2016). doi:10.1038/ncomms12942
Author:         Ulschan Scheler
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       Item: 
        Title: Results
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        Children: 
         Item: 
          Title: Identification of HFSs
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       Item: 
        Title: Figure™1Biosynthesis of PDs in Rosmarinus officinalis and sage species.State of the knowledge on the biosynthetic pathway from GGPP to CA, CO and PA before this study. The terpene synthases CPS and MiS cyclize GGPP to miltiradiene, which spontaneously oxi
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         Item: 
          Title: Triple-mutants of FS recapitulate HFS activity
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       Item: 
        Title: Figure™2Homology modelling-based mutagenesis of FS and HFS.(a) Part of the GC-MS profile of hexane extracts from rosemary leaf surfaces and from yeast strains co-expressing GGPPS, CPS, MS, ATR1 and indicated CYPs (selected msolz signals: 270, 272, 286, 30
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       Item: 
        Title: Table 1 
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        Children: 
         Item: 
          Title: C20 oxidation by CYP76AK6-8 leads to CA
          Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431
         Item: 
          Title: In vivo quantification of pathway products by NMR spectroscopy
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         Item: 
          Title: Phylogeny of CYP76AH and CYP76AK proteins
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       Item: 
        Title: Methods
        Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71
        Children: 
         Item: 
          Title: Plant material
          Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1b26f7b2
         Item: 
          Title: Chemicals
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         Item: 
          Title: Tissue collection and RNA preparation for quantitative RT-PCR
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         Item: 
          Title: Quantitative real-time PCR analysis
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         Item: 
          Title: Construction of golden gate compatible yeast expression MoClo vectors
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       Item: 
        Title: Figure™3Functional analysis of C20Ox.(a) Expression profile of CYP76AK6-8 in young and old trichomes and leaves without trichomes. CYP76AK6 is from S. fruticosa, CYP76AK7-8 are from R. officinalis. Quantitative reverse transcriptase-PCR data were obtained
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        Children: 
         Item: 
          Title: Designing synthetic galactose-inducible promoters
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         Item: 
          Title: Isolation and cloning of genes
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          Title: Phylogenetic analysis
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       Item: 
        Title: Figure™4Overview of the engineered CA pathway.FS (CYP76AH1) oxidizes abietatriene to ferruginol, whereas HFS (CYP76AH22-24) are able to further oxidize ferruginol to 11-hydroxyferruginol. 11-hydroxyferruginol spontaneously oxidizes to hydroxyferruginol qu
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        Children: 
         Item: 
          Title: Site-directed mutagenesis
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          Title: Yeast microsome isolation
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          Title: In vitro CYP assays
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         Item: 
          Title: Transient expression in N. benthamiana
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         Item: 
          Title: Homology modelling
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         Item: 
          Title: Production and quantification of diterpenes in yeast
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          Title: GC-MS analysis
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         Item: 
          Title: RP-UPLC-ESI-MSsolMS analysis (LC-MS)
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         Item: 
          Title: Isolation of new PDs and structure confirmation by NMR
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         Item: 
          Title: Data availability
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       Item: 
        Title: KingstonD. G. I.Taxol, a molecule for all seasonsChem. Commun.108678802001WattsG.Nobel awarded to discoverers of ivermectin and artemisininBMJ351h53522015JordánM. J.LaxV.RotaM. C.LoránS.SotomayorJ. A.Relevance of carnosic acid, carnosol, and rosmarinic ac
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       Item: 
        Title: We thank Jürgen Schmidt for assistance in the analysis of mass spectral data and Benedikt Athmer for his support concerning phylogenetic analysis. We also thank Arianne Schnabel and Sarah Vorndran for technical assistance. The work was supported in part b
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        Title: ACKNOWLEDGEMENTS
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        Title: Author contributions
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       Item: 
        Title: Additional information
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