Development of Trichoderma reesei transformation technology with the aid of an episomal vector system

    Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

    Abstract

    In conventional transformation procedures of filamentous fungi the transformation frequencies have been modest and only integrative vectors have been available. In modern fungal molecular biology, however, screening approaches with the help episomal vectors have advanced the progress of the field greatly. For example, in enzyme discovery or protein engineering it is beneficial to be able to screen for new enzymes or improved mutant enzymes in the ultimate production host of the enzyme. Episomal vector systems have been developed for a few fungal species including the Aspergilli, but Trichoderma reesei has suffered from a lack of such a system. We have developed T. reesei transformation technology for genetic and enzyme screening purposes. The transformation procedure was optimised to improve the transformation frequencies of any vectors. Telomeric episomal vectors were used, and with these we obtained transformation frequencies that are far above those obtained with integrative vectors and are feasible for genetic and enzyme screening purposes. Procedures for plasmid rescue, colony picking and microtiter plate cultivation were also developed. The use of the novel transformation system in enzyme and genetic screening experiments will be discussed.
    Original languageEnglish
    Title of host publication3rd European Federation of Biotechnology Conference
    Subtitle of host publicationPhysiology of Yeasts and Filamentous Fungi PYFF3
    Place of PublicationEspoo
    PublisherVTT Technical Research Centre of Finland
    Pages149
    ISBN (Electronic)978-951-38-6314-2
    ISBN (Print)978-951-38-6313-5
    Publication statusPublished - 2007
    Event3rd European Federation of Biotechnology Conference : Physiology of Yeasts and Filamentous Fungi - Helsinki, Finland
    Duration: 13 Jun 200716 Jun 2007

    Publication series

    SeriesVTT Symposium
    Number245
    ISSN0357-9387

    Conference

    Conference3rd European Federation of Biotechnology Conference
    Abbreviated titlePYFF3
    CountryFinland
    CityHelsinki
    Period13/06/0716/06/07

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