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      Promoter-tagged restriction enzyme-mediated insertion (PT-REMI) mutagenesis in Aspergillus niger.

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      Journal: Molecular & general genetics : MGG
      Keywords: Amino Acid Sequence, Aspergillus niger, cytology, genetics, Base Sequence, Blotting, Northern, Blotting, Southern, Color, DNA Restriction Enzymes, metabolism, Electron Transport Complex IV, Gene Expression Regulation, Fungal, Genetic Markers, Glucan 1,4-alpha-Glucosidase, Molecular Sequence Data, Multienzyme Complexes, Mutagenesis, Insertional, Plasmids, Promoter Regions, Genetic, Sequence Homology, Amino Acid, Spores, Fungal, Transformation, Genetic

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          Abstract

          Promoter-tagged restriction enzyme-mediated insertion (PT-REMI) mutagenesis was performed in the fungus Aspergillus niger, using a plasmid containing a strong transcriptional promoter. Two DNA-tagged mutants were analyzed in detail. A white-spored mutant was shown to contain a plasmid insertion that disrupted a gene that shows a high degree of homology to the polyketide synthase gene wA of A. nidulans. A morphological mutant was shown to contain a plasmid insertion in the 5' upstream region of a gene that strongly resembles COX5, which encodes the cytochrome c oxidase subunit V. Insertion of the plasmid resulted in enhanced expression of the COX5 RNA, demonstrating that the combination of REMI with a promoter-containing insert can be used to activate gene transcription.

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          PubMed ID:: 10503533

          ScienceOpen disciplines: Chemistry
          Keywords: Amino Acid Sequence,Aspergillus niger,cytology,genetics,Base Sequence,Blotting, Northern,Blotting, Southern,Color,DNA Restriction Enzymes,metabolism,Electron Transport Complex IV,Gene Expression Regulation, Fungal,Genetic Markers,Glucan 1,4-alpha-Glucosidase,Molecular Sequence Data,Multienzyme Complexes,Mutagenesis, Insertional,Plasmids,Promoter Regions, Genetic,Sequence Homology, Amino Acid,Spores, Fungal,Transformation, Genetic
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          ScienceOpen disciplines: Chemistry
          Keywords: Amino Acid Sequence, Aspergillus niger, cytology, genetics, Base Sequence, Blotting, Northern, Blotting, Southern, Color, DNA Restriction Enzymes, metabolism, Electron Transport Complex IV, Gene Expression Regulation, Fungal, Genetic Markers, Glucan 1,4-alpha-Glucosidase, Molecular Sequence Data, Multienzyme Complexes, Mutagenesis, Insertional, Plasmids, Promoter Regions, Genetic, Sequence Homology, Amino Acid, Spores, Fungal, Transformation, Genetic

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