Title
Functional elements in the promoter region of the Aspergillus nidulans gpdA gene encoding glyceraldehyde-3-phosphate dehydrogenase
Author
Medisch Biologisch Laboratorium TNO
Punt, P.J.
Dingemanse, M.A.
Kuyvenhoven, A.
Soede, R.D.M.
Pouvels, P.H.
van den Hondel, C.A.M.J.J.
Publication year
1990
Abstract
Analysis of the promoter region of the highly expressed Aspergillus nidulans gpdA gene is described. The nucleotide (nt) sequence of a 1.3-kb region upstream from the ATG was determined. Comparison with promoter regions of other Aspergillus and Neurospora genes revealed several regions similar sequence. Both random and site-specific mutations were introduced into the promoter region of the gpdA gene, and the resulting mutant promoters were fused to the Escherichia coli lacZ gene. The constructed fusions were introduced into A. nidulans and transformants that contained one copy of these fusions at the argB locus were analysed. β-Galactosidase assays and primer extension experiments were used to identify sequence elements involved in transcription activation and transcription initiation. Two elements, located around 650 and 250 nt upstream from the major transcription start point (tsp), were identified as transcription activation elements. These elements coincide with regions of putative secondary structure (direct or inverted repeats). A third element, a C + T-rich region directly upstream from the major tsp, was shown to be involved in correct initiation of transcription.
Subject
Aspergillus niger
LacZ fusion gene
Recombinant DNA
Site-directed mutagenesis
Targeted single-copy integration
Transcription activation
Transcription start point
Glyceraldehyde 3 phosphate dehydrogenase
Aspergillus nidulans
DNA flanking region
Fungus
Gene fusion
Genetic engineering
Heredity
Nonhuman
Nucleotide sequence
Priority journal
Promoter region
Aspergillus nidulans
Base Sequence
beta-Galactosidase
Cloning, Molecular
DNA Mutational Analysis
Gene Expression Regulation, Fungal
Glyceraldehyde-3-Phosphate Dehydrogenases
Molecular Sequence Data
Oligonucleotide Probes
Promoter Regions (Genetics)
Regulatory Sequences, Nucleic Acid
Restriction Mapping
Transcription, Genetic
Aspergillus
Aspergillus niger
Emericella nidulans
Escherichia coli
Fungi
Neurospora
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DOI
https://doi.org/10.1016/0378-1119(90)90142-e
TNO identifier
231261
ISSN
0378-1119
Source
Gene, 93 (93), 101-109
Document type
article