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    Plant Mol Biol. 1993 May;22(2):379-83.

    Isolation of cytochrome P-450 cDNA clones from the higher plant Catharanthus roseus by a PCR strategy.

    Meijer AH, Souer E, Verpoorte R, Hoge JH.

    Centre for Bio-Pharmaceutical Sciences, Leiden University, Gorlaeus Laboratories, Netherlands.

    Cytochrome P-450 monooxygenases are membrane-bound enzymes involved in a wide range of biosynthetic pathways in plants. An efficient PCR strategy for isolating cytochrome P-450 cDNA clones from plant cDNA libraries is described. A set of degenerate primers for PCR amplification was designed to recognize nucleotide sequences specifying the highly conserved haembinding region of cytochrome P-450 proteins. Using this primer set and a non-specific primer, complementary to either the poly(A) tail of the cDNA clones or a phage vector sequence, we isolated 16 different cytochrome P-450 cDNA sequences from a cDNA library of Catharanthus roseus.

    PMID: 8507838 [PubMed - indexed for MEDLINE]

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