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  • 1
    ISSN: 1432-0983
    Keywords: Key words  Agaricus bisporus ; Homologous recombination ; Exo-β-1 ; 3-glucanase ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract   Homologous integration was studied in the common mushroom, Agaricus bisporus, using a plasmid (pHAG3-1) carrying the hygromycin-resistance gene and a 3.2-kb genomic fragment from A. bisporus. Homologous integration was found in 30–60% of the transformants obtained with pHAG3-1 linearized at three different positions within the homologous sequence, generating either blunt, 5′- or 3′-protruding ends. The genomic fragment was found to contain two homologous open reading frames in tandem, which showed 60% similarity to exo-β-1,3-glucanases from Saccharomyces cerevisiae and Candida albicans. The level of the corresponding mRNA is low in the vegetative mycelium and relatively high in fruiting bodies. In the vegetative mycelium of a transformant with tandemly integrated pHAG3-1 plasmids at the homologous position, exoglucanase mRNA was strongly increased without any apparent effect on growth rate or morphology.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5028
    Keywords: β-1,3-glucanase ; organ-specific expression ; pathogenesis-related proteins ; regulatory elements ; tobacco mosaic virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Infection of tobacco by tobacco mosaic virus (TMV) induces coordinate expression of genes encoding acidic and basic β-1,3-glucanase isoforms. These genes are differentially expressed in response to other treatments. Salicylate treatment induces acidic glucanase mRNA to a higher level than basic glucanase mRNA. Ethylene treatment and wounding strongly induce the basic glucanase genes but have little effect on genes encoding the acidic isoforms. Furthermore, the basic glucanase genes are constitutively expressed in roots and lower leaves of healthy plants, whereas the acidic glucanase genes are not. In order to investigate how these expression patterns are established, we fused promoter regions of an acidic and a basic glucanase gene to the β-glucuronidase (GUS) reporter gene and examined expression of these constructs in transgenic tobacco plants. A fragment of 1750 bp and two 5′-truncated fragments of 650 bp and 300 bp of the acidic glucanase promoter were tested for induction of GUS gene expression after salicylate treatment and TMV infection. Upstream sequences of 1750 bp and 650 bp were sufficient for induction of the reporter gene by salicylate treatment and TMV infection, but the activity of the 300 bp fragment was strongly reduced. The results suggest that the 1750 bp upstream sequence of the acidic glucanase gene contains multiple regulatory elements. For the basic glucanase promoter it is shown that 1476 bp of upstream sequences were able to drive expression in response to TMV infection and ethylene treatment, but no response was found to incision wounding. Furthermore, high GUS activity was found in lower leaves and roots of healthy transgenic plants, carrying the 1476 bp basic glucanase promoter/GUS construct. When the promoter was truncated up to position −446 all activity was lost, indicating that the region between −1476 and −446 of the basic glucanase promoter is necessary for organ-specific and developmentally regulated expression as well as for induced expression in response to infection and other stress treatments.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 18 (1992), S. 155-158 
    ISSN: 1573-5028
    Keywords: pathogenesis-related proteins ; tobacco mosaic virus infection ; cis-regulatory elements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract cis-Regulatory elements involved in tobacco mosaic virus (TMV)-inducible expression were indentified in a tobacco PR-5 gene, encoding an acidic thaumatin-like protein. By fusing upstream sequences of the PR-5 gene to the GUS reporter gene and analysing transgenic plants containing these fusions for local and systemic induction of GUS activity by TMV, it was found that sequences between-1364 and-718 are involved in TMV induction of PR-5 gene expression.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1617-4623
    Keywords: Key words Agaricus bisporus ; Transformation ; Edible mushrooms ; Hygromycin B phosphotransferase ; Homobasidiomycetes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Application of biotechnology to the cultivated mushroom, Agaricus bisporus, has been hampered thus far by the lack of a transformation system. Here, transformation of both a homo- and a heterokaryotic strain of A. bisporus to hygromycin B resistance is described. Transforming DNA was integrated into the A. bisporus genome and stably maintained throughout vegetative growth. Transformants of the heterokaryotic strain formed transgenic fruiting bodies. Promoters derived from the unrelated ascomycete Aspergillus nidulans and from A. bisporus itself, were able to drive expression of the hygromycin B resistance gene. Expression controlled by a fragment of 265 bp from the A. bisporus GPD promoter was sufficient to generate transformants. However, transformation efficiency was not enhanced by using this homologous promoter.
    Type of Medium: Electronic Resource
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