Functional elements in the promoter region of the Aspergillus nidulans gpdA gene encoding glyceraldehyde-3-phosphate dehydrogenase
article
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.
Topics
Aspergillus nigerLacZ fusion geneRecombinant DNASite-directed mutagenesisTargeted single-copy integrationTranscription activationTranscription start pointGlyceraldehyde 3 phosphate dehydrogenaseAspergillus nidulansDNA flanking regionFungusGene fusionGenetic engineeringHeredityNonhumanNucleotide sequencePriority journalPromoter regionAspergillus nidulansBase Sequencebeta-GalactosidaseCloning, MolecularDNA Mutational AnalysisGene Expression Regulation, FungalGlyceraldehyde-3-Phosphate DehydrogenasesMolecular Sequence DataOligonucleotide ProbesPromoter Regions (Genetics)Regulatory Sequences, Nucleic AcidRestriction MappingTranscription, GeneticAspergillusAspergillus nigerEmericella nidulansEscherichia coliFungiNeurospora
TNO Identifier
231261
ISSN
03781119
Source
Gene, 93(1), pp. 101-109.
Pages
101-109
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