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Approaches to isolating a cDNA encoding thebaine synthase of morphine
cDNA library screening and Gene expression
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Secondary metabolism (also called specialized metabolism) is a term for pathways and molecule products of metabolism that are not absolutely required for the survival of the organism. Examples of the products include antibiotics and pigments. To distinguish non-secondary ("ordinary") metabolism, the term basic metabolism is sometimes used. Secondary metabolites are produced by many microbes, plants, fungi and animals. This study was focusing on the molecular characterization and further in vitro biochemical analysis of thebaine synthase from opium poppy Papaver somniferum using the yeast two-hybrid assay. To this end, a salutaridinol 7-O-acetyltransferase-human gene product (hSos), "Bait", fusion was generated and screened against a P. somniferum cDNA library cloned into the activation domain vector pMyr "Prey". Salutaridinol 7-O-acetyltransferase of morphine biosynthesis was initally identified and isolated as a cDNA clone that corresponds to the internal amino acid sequences of the native enzyme purified from a cell suspension culture of P. somniferum proteins.
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