Transformation system for hypocrea jecorina (trichoderma reesei) that favors homologous integration and employs reusable bidirectionally selectable markers

Matthias G. Steiger, Marika Vitikainen, Pekka Uskonen, Kurt Brunner, Gerhard Adam, Tiina Pakula, Merja Penttilä, Markku Saloheimo, Robert L. Mach, Astrid R. Mach-Aigner (Corresponding Author)

    Research output: Contribution to journalArticleScientificpeer-review

    97 Citations (Scopus)

    Abstract

    Hypocrea jecorina is an industrially important filamentous fungus due to its effective production of hydrolytic enzymes. It has received increasing interest because of its ability to convert lignocellulosic biomass to monomeric sugars, which can be converted into biofuels or platform chemicals. Genetic engineering of strains is a highly important means of meeting the requirements of tailor-made applications. Therefore, we report the development of a transformation system that allows highly efficient gene targeting by using a tmus53 (human LIG4 homolog) deletion strain. Moreover, it permits the unlimited reuse of the same marker by employing a Cre/loxP-based excision system. Both marker insertion and marker excision can be positively selected for by combining resistance to hygromycin B and loss of sensitivity to fluoroacetamide. Finally, the marker pyr4, also positively selectable for insertion and loss, can be used to remove the cre gene.

    Original languageEnglish
    Pages (from-to)114-121
    JournalApplied and Environmental Microbiology
    Volume77
    Issue number1
    DOIs
    Publication statusPublished - 1 Jan 2011
    MoE publication typeA1 Journal article-refereed

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