Print Email Facebook Twitter The effect of multiple copies of the upstream region of the Aspergillus niger glucoamylase-encoding gene on expression Title The effect of multiple copies of the upstream region of the Aspergillus niger glucoamylase-encoding gene on expression Author Verdoes, J.C. Punt, P.J. Stouthamer, A.H. van den Hondel, C.A.M.J.J. Medisch Biologisch Laboratorium TNO Centraal Instituut voor Voedingsonderzoek TNO Publication year 1994 Abstract The regulation of transcription of the glucoamylase-encoding gene (glaA) of Aspergillus niger was studied. To facilitate this study a reporter strain containing a fusion of the glaA promoter (P(glaA)) of A. niger to the β- glucuronidase-encoding gene (uidA) of Escherichia coli was constructed. To analyze whether regulatory proteins are involved in the regulation of glaA, multiple copies of P(glaA) were introduced into this reporter strain. Analysis of the resulting strains revealed that introduction of an increasing number of P(glaA) copies resulted in lower expression of the uidA reporter gene and the endogenous glaA gene in cultures cultivated on different inducing carbon sources. However, repression by xylose was not influenced by the copy number of P(glaA). These results indicate that the expression of genes under control of P(glaA) are regulated by specific trans-acting regulatory protein(s). Deletion analysis of P(glaA) indicated that regulatory proteins interact with DNA sequences within 0.5-kb upstream from the ATG, whereas sequences between about 0.8- and 0.5-kb upstream from the ATG are required for high-level expression of glaA. Molecular Sequence Numbers: GENBANK: Z30918; Chemicals/CAS: Carbohydrates; DNA-Binding Proteins; Genetic Vectors; Glucan 1,4-alpha-Glucosidase, EC 126.96.36.199; RNA, Messenger; Xylose Subject β-GlucuronidaseActivatorCatabolite repressionDeletion analysisExpressionTranscription regulationBeta glucuronidaseGlucan 1,4 alpha glucosidaseRegulator proteinCatabolite repressionDNA flanking regionDNA sequenceEnzyme activationEscherichia coliGene deletionGene expressionNonhumanPriority journalReporter geneTranscription regulationAspergillus nigerBase SequenceCarbohydratesComparative StudyDNA Mutational AnalysisDNA-Binding ProteinsEnzyme InductionEnzyme RepressionGene Expression Regulation, EnzymologicGene Expression Regulation, FungalGenes, FungalGenes, ReporterGenetic VectorsGlucan 1,4-alpha-GlucosidaseMolecular Sequence DataMultigene FamilyPromoter Regions (Genetics)RNA, MessengerSequence DeletionSequence Homology, Nucleic AcidSupport, Non-U.S. Gov'tXyloseEscherichia coli To reference this document use: http://resolver.tudelft.nl/uuid:12ce2d2e-6774-4378-b29b-56a7cf2629b8 DOI https://doi.org/10.1016/0378-1119(94)90003-5 TNO identifier 82312 Source Gene, 145, 179-187 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.