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    Title: An Efficient System for Heterologous Expression of Secondary Metabolite Genes in Aspergillus nidulans
    Authors: Chiang, Yi-Ming
    Oakley, C. Elizabeth
    Ahuja, Manmeet
    Entwistle, Ruth
    Schultz, Aric
    Chang, Shu-Lin
    Sung, Calvin T.
    Wang, Clay C. C.
    Oakley, Bed R.
    Contributors: 藥學系
    Keywords: Polyketide Synthase Gene
    Combinatorial Chemistry
    Date: 2013-05
    Issue Date: 2014-05-26 10:43:03 (UTC+8)
    Publisher: Amer Chemical Soc
    Abstract: Fungal secondary metabolites (SMs) are an important source of medically valuable compounds. Genome projects have revealed that fungi have many SM biosynthetic gene clusters that are not normally expressed. To access these potentially valuable, cryptic clusters, we have developed a heterologous expression system in Aspergillus nidulans. We have developed an efficient system for amplifying genes from a target fungus, placing them under control of a regulatable promoter, transferring them into A. nidulans, and expressing them. We have validated this system by expressing nonreducing polyketide synthases of Aspergillus terreus and additional genes required for compound production and release. We have obtained compound production and release from six of these nonreducing polyketide synthases and have identified the products. To demonstrate that the procedure allows transfer and expression of entire secondary metabolite biosynthetic pathways, we have expressed all the genes of a silent A terreus cluster and demonstrate that it produces asperfuranone. Further, by expressing the genes of this pathway in various combinations, we have clarified the asperfuranone biosynthetic pathway. We have also developed procedures for deleting entire A nidulans SM clusters. This allows us to remove clusters that might interfere with analyses of heterologously expressed genes and to eliminate unwanted toxins.
    Relation: Journal of the American Chemical Society, v.135 n.20 pp.7720-7731
    Appears in Collections:[Dept. of Pharmacy] Periodical Articles

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