Promoter-tagged restriction enzyme-mediated insertion (PT-REMI) mutagenesis in Aspergillus niger.
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.
Shuster
J R
JR
Novo Nordisk Biotech Inc, Davis, CA 95616, USA.
Bindel Connelley
M
M
D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000166:cytology / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003116:Color; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D003576:Electron Transport Complex IV / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D005819:Genetic Markers; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics; D016254:Mutagenesis, Insertional; D010957:Plasmids; D011401:Promoter Regions, Genetic; D017386:Sequence Homology, Amino Acid; D013172:Spores, Fungal / Q000235:genetics; D014170:Transformation, Genetic
D005819:Genetic Markers; D009097:Multienzyme Complexes; D003576:Electron Transport Complex IV; D004262:DNA Restriction Enzymes; D005087:Glucan 1,4-alpha-Glucosidase