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 3.2.1.3; RNA, Messenger; Xylose
Subject
β-Glucuronidase
Activator
Catabolite repression
Deletion analysis
Expression
Transcription regulation
Beta glucuronidase
Glucan 1,4 alpha glucosidase
Regulator protein
Catabolite repression
DNA flanking region
DNA sequence
Enzyme activation
Escherichia coli
Gene deletion
Gene expression
Nonhuman
Priority journal
Reporter gene
Transcription regulation
Aspergillus niger
Base Sequence
Carbohydrates
Comparative Study
DNA Mutational Analysis
DNA-Binding Proteins
Enzyme Induction
Enzyme Repression
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Fungal
Genes, Fungal
Genes, Reporter
Genetic Vectors
Glucan 1,4-alpha-Glucosidase
Molecular Sequence Data
Multigene Family
Promoter Regions (Genetics)
RNA, Messenger
Sequence Deletion
Sequence Homology, Nucleic Acid
Support, Non-U.S. Gov't
Xylose
Escherichia coli
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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