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Fungal Transformation: From Protoplasts to Targeted Recombination Systems

  • Juan F. MartínEmail author
Chapter
Part of the Fungal Biology book series (FUNGBIO)

Abstract

Transformation of fungi is required for understanding their molecular biology and for manipulation of strains of industrial interest. The development of efficient transformation systems has been hampered by the extreme diversity of ascomycetes, basidiomycetes, and zygomycetes. Polyethylene glycol-assisted transformation of fungal protoplasts is the most widely used system but alternative procedures, e.g., electroporation and Agrobacterium tumefaciens-mediated transformation have been developed.

Most transformation procedures rely on integrative vectors, but AMA1-containing vectors that replicate autonomously are available. Exogenous DNA integrates frequently at nonhomologous sites in the genome. This is a problem for targeted gene disruption and for quantifying gene expression. Procedures for directed integration of a single plasmid copy at a specific locus in the genome have been developed.

Mutants of different fungi impaired in the NHEJ system allowed to select recombinants at homologous sites with high efficiency. These advances have contributed significantly to the progress in the molecular biology of fungi.

Keywords

Fungal transformation Selective markers AMA1-replicating plasmids Agrobacterium-mediated transformation Homologous and nonhomologous recombination Targeted integration Nonhomologous end joining 

Notes

Acknowledgements

I acknowledge the help of Dr. Paloma Liras for valuable scientific discussions on the manuscript and the initial scientific collaboration on fungal transformation of Drs. J.M. Cantoral, J.L. Barredo, B. Díez, S. Gutiérrez, F. Fierro, J. Casqueiro, O. Bañuelos, G. Liu and K. Kosalková.

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© Springer International Publishing Switzerland 2015

Authors and Affiliations

  1. 1.Department of Molecular Biology, Faculty of Biology and Environmental ScienceUniversity of LeónLeónSpain

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