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  • 1
    ISSN: 1432-0983
    Keywords: A. niger trpC gene ; Sequence analysis ; Amino acid homology ; Conservation ; Corrected A. nidulans trpC DNA sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleotide sequence of the Aspergillus niger tryptophan C (trpC) gene was determined. Northern hybridization and S1-mapping experiments showed the presence of a 2.6 kb trpC poly(A)+ RNA with two very short (5 and 6 nucleotides) noncoding 5′-regions. Comparison of the predicted amino acid sequence with that of trp gene proteins of pro- and eukaryotic organisms revealed three functional domains (G, C, F) in the A. niger TrpC protein which catalyse the glutarnine amidotransferase reaction (GAT), the indole-3-glycerol phosphate synthase reaction (IGPS) and the N-(5′-phosphoribosyl) anthranilate isomerase reaction (PRAI), respectively. These domains are highly conserved and bordered by short areas showing less homology. Within the F domain of the trpC gene in A. niger, A. nidulans and Neurospora crassa, a region encoding 30 amino acids was found which is absent in the analogous genes of Saccharomyces cerevisiae and prokaryotic organisms. This region has features of a mutated in-phase intron.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-9368
    Keywords: transcription regulation ; regulatory protein ; amdS selection marker ; cosmid vector ; CHEF analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A strategy, based on the usage of theamdS selection marker and a cosmid vector containing four copies of the glucoamylase gene (glaA), was developed to obtain glucoamylase (GLA)-overproducingA. niger strains. With this strategy, fungal strains carrying up to 200 copies of theglaA gene could be isolated at a relatively high frequency. In each transformant analysed, integration occurred in a single chromosome. A significant increase in the extracellular GLA production was observed in most of the transformants carrying multiple copies of theglaA gene. Further analysis showed that the amount of GLA that is produced was not proportional to the number ofglaA copies in these transformants. However, the level of GLA production clearly correlated with the amount ofglaA mRNA produced in these transformants. From these results it is concluded that GLA production is limited at the level of transcription.
    Type of Medium: Electronic Resource
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