Agrobacterium tumefaciens-Mediated Transformation

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    Abstract

    The use of Agrobacterium tumefaciens-mediated transformation for achieving genetic transformation of fungi has steadily increased over the last decade, and has proven to be almost universally applicable technique once suitable selection markers have been developed. In recent years the major technical advances has been made within the initial steps of the process, more specifically the efficient construction of plasmids for performing targeted genome modifications. This chapter provides a generic protocol for performing genetic transformation of ascomycetes via A. tumefaciens-mediated transformation (AMT) and guidelines for optimizing the AMT process with new fungal species. The chapter also includes a highly efficient vector construction system based on Uracil Specific Excisions Reagent (USER) cloning and specific PCR generated building blocks, which can be combined ad hoc to create complex plasmids in a single cloning step.
    Original languageEnglish
    Title of host publicationGenetic Transformation Systems in Fungi
    EditorsMarco A. van den Berg, Karunakaran Maruthachalam
    Number of pages20
    Volume1
    PublisherSpringer
    Publication date2015
    Pages143-162
    Chapter14
    ISBN (Print)978-3-319-10141-5
    ISBN (Electronic)978-3-319-10142-2
    DOIs
    Publication statusPublished - 2015
    SeriesFungal Biology
    ISSN2198-7777

    Keywords

    • Developmental biology
    • Plant genetics & genomics
    • Plant pathology
    • Plant breeding/biotechnology
    • Systems biology

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