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H van den Broek

Publications and source records attributed to H van den Broek.

11 recordsLinked to original sources
PubMed1995

Isolation and characterization of the glucose-6-phosphate dehydrogenase encoding gene (gsdA) from Aspergillus niger.

Genomic and cDNA clones encoding glucose-6-phosphate dehydrogenase (G6PD) were isolated from the fungus Aspergillus niger. Sequence analysis of the glucose-6-phosphate dehydrogenase gene (gsdA) revealed an open reading frame of 1530 bp, encoding a protein of 58,951 kDa. The gsdA gene is interrupted by nine introns the most proximal of which is exceptionally large (348 bp). The region upstream of the ATG contains several C+T-rich stretches. The two major and one minor transcription start points are all located within these regions. In the upstream region several direct and inverted repeats, but no clear TATA or CCAAT boxes can be found. A. niger strains overproducing G6PD were constructed by cotransformation of gsdA subclones. Overexpression of G6PD was shown to be deleterious for the fungus, especially when cotransformants were grown on media containing ammonia. Attempts to construct a gsdA null mutant by gene disruption were unsuccessful.

Amino Acid Sequence↗
PubMed1989

Tryptophan auxotrophic mutants in Aspergillus niger: inactivation of the trpC gene by cotransformation mutagenesis.

Aspergillus niger tryptophan auxotrophic mutants have been isolated after UV irradiation of conidiospores. The mutants belong to two different complementation groups, trpA and trpB, which complement each other in heterokaryons. Neither of the mutations could be complemented with the cloned A. niger trpC gene. To obtain A. niger trpC mutants in a direct way, gene inactivation by cotransformation was performed. For this purpose an in-frame gene fusion between the A. niger trpC and Escherichia coli lacZ genes was constructed and shown to be functionally expressed after introduction into A. niger by cotransformation with the pyrA gene as selective marker. Among the beta-galactosidase expressing cotransformants, obtained with either circular or linearized vectors, no trpC mutants were detected, even after enrichment. Such mutants, however, could be obtained by cotransformation of A. niger with specific fragments of the fusion gene. Biochemical analysis of the cotransformants indicated that in nearly all cases the fusion gene had replaced the wild-type trpC gene. Genetic analysis showed that the trpC mutation is not linked to any of the A. niger loci described so far. The trpC mutants can be complemented by the cloned A. niger trpC gene as well as by the A. nidulans trpC gene.

Aspergillus niger↗
PubMed1988

Isolation and transformation of the pyruvate kinase gene of Aspergillus nidulans.

The Aspergillus nidulans pyruvate kinase gene was isolated by heterologous hybridization using the corresponding yeast gene as a probe. A 2.9 kb EcoRI/BamHI fragment, which exclusively hybridized to the yeast gene, was subcloned in pBR322. This clone was used to transform an A. nidulans pkiA deletion mutant to PKI+. The analysis of transformants with respect to the kind of integration revealed about 80% homologous integration--55% by a double cross-over event (type III integration), 25% by a single cross-over event (type I integration). Type II transformants (20%) that arise by non-homologous integration have not been further characterized with respect to the sites of integration. A direct correlation between the number of copies of the gene integrated into the genome and the measured pyruvate kinase activity was found after growth ona glycolytic carbon source. From this, it was concluded that the 2.9 kb EcoRI/BamHI fragment contains the complete pyruvate kinase structural gene, including the promoter region. However, after growth on a gluconeogenic carbon source, the regulation of gene expression was found to be disturbed. On acetate an increase in activity per gene copy (0.2 IU) was found in the transformants, as compared with wild-type levels. It is suggested that the pyruvate kinase gene is regulated by negative control, and that some sequences involved in this regulation are missing in the cloned fragment.

Aspergillus nidulans↗
PubMed1982

Immunopathology of psoriasis: a comparison with other parakeratotic lesions.

Characteristic in vivo binding of IgG and C3 to the stratum corneum of psoriatic lesions has been implicated as a significant event in the pathogenesis of the disease. Immunofluorescent findings of psoriatic lesions are compared with other parakeratotic and nonparakeratotic lesions. Parakeratotic lesions studied include verrucae vulgares in patients with psoriasis, verrucae vulgares in patients without psoriasis, actinic keratoses and lichen simplex chronicus. Psoriatic lesions and other parakeratotic lesions demonstrate similar immunopathological phenomena within the involved stratum corneum: (1) A high percentage of the lesions stain positively with one or more of the immunoglobulins (IgG, IgM and IgA). (2) No predominance of any immunoglobulin class is found. (3) The predominant sites of immunoglobulin deposition are within the regions of parakeratosis. (4) C3 deposition is found in a high percentage of the lesions and is present at the sites of immunoglobulin binding. By contrast only a small percentage of nonparakeratotic lesions display stratum corneum fluorescence. These data are consistent with macromolecular leakage through the lesional microvasculature and high affinity binding of immunoglobulins by parakeratotic stratum corneum. Similar immunofluorescent findings in psoriatic lesions and nonpsoriatic parakeratotic lesions suggest that immunoglobulin binding in psoriatic lesional stratum corneum is not a significant event in the pathogenesis of psoriatic lesions.

Antigen-Antibody Reactions↗
PubMed1980

Treatment of prurigo nodularis with thalidomide.

A 57-year-old man with prurigo nodularis, was unresponsive to conventional therapy, was successfully treated with thalidomide. The pharmacology and side effects of the drug are briefly reviewed. Why thalidomide is effective for prurigo nodularis is unclear, but possible mechanisms of action are discussed.

Humans↗