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Agrobacterium tumefaciens-Mediated Transformation

  • Rasmus John Normand FrandsenEmail author
Chapter
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Part of the Fungal Biology book series (FUNGBIO)

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.

Keywords

Agrobacterium tumefaciens A. tumefaciens-mediated transformation (AMT) Uracil Specific Excisions Reagent (USER) cloning USER-Bricks AMT experiments AMT process 

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Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  1. 1.Department of Systems Biology, Group for Eukaryotic Molecular Cell BiologyTechnical University of DenmarkLyngbyDenmark

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