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Agrobacterium-mediated transformation as a tool for functional genomics in fungi

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Abstract

In the era of functional genomics, the need for tools to perform large-scale targeted and random mutagenesis is increasing. A potential tool is Agrobacterium-mediated fungal transformation. A. tumefaciens is able to transfer a part of its DNA (transferred DNA; T-DNA) to a wide variety of fungi and the number of fungi that can be transformed by Agrobacterium-mediated transformation (AMT) is still increasing. AMT has especially opened the field of molecular genetics for fungi that were difficult to transform with traditional methods or for which the traditional protocols failed to yield stable DNA integration. Because of the simplicity and efficiency of transformation via A. tumefaciens, it is relatively easy to generate a large number of stable transformants. In combination with the finding that the T-DNA integrates randomly and predominantly as a single copy, AMT is well suited to perform insertional mutagenesis in fungi. In addition, in various gene-targeting experiments, high homologous recombination frequencies were obtained, indicating that the T-DNA is also a useful substrate for targeted mutagenesis. In this review, we discuss the potential of the Agrobacterium DNA transfer system to be used as a tool for targeted and random mutagenesis in fungi.

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Acknowledgements

We thank R. Cardoza, M. Challen, B. Donzelli, S. Gutierrez, T. Heinekamp, H. Khanh, Y. Lee, C. Rogers, J. Skov, J. Kan, and K. Welzel for their consent to use their data that were presented at the 21st and 22nd FGC (Asilomar, USA), at ECFG6 (Pisa, Italy), and/or at ECFG7 (Copenhagen, Denmark). We thank V. Garre, G. Gay, J. Gomez-Mateo, S. Covert, M. Rep, M. Furlaneto, K. Plummer, S. Kang, A. Idnurm, D. Gardiner, J.F. Martin, and P. Tudzynski for sharing their unpublished data and discussion. We also thank Patricia vanKuyk and Jaap Visser for critically reading this manuscript.

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  1. Caroline B. Michielse

    Present address: Department Human Genetics, University Medical Centre Nijmegen, P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands

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  1. Institute of Biology, Clusius Laboratory, Fungal Genetics Research Group, Leiden University, Wassenaarseweg 64, 2333 AL, Leiden, The Netherlands

    Caroline B. Michielse, Paul J. J. Hooykaas, Cees A. M. J. J. van den Hondel & Arthur F. J. Ram

  2. TNO Quality of Life, Microbiology, Utrechtseweg 48, 3500 AJ, Zeist, The Netherlands

    Cees A. M. J. J. van den Hondel & Arthur F. J. Ram

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  1. Caroline B. Michielse
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Correspondence to Arthur F. J. Ram.

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Michielse, C.B., Hooykaas, P.J.J., van den Hondel, C.A.M.J.J. et al. Agrobacterium-mediated transformation as a tool for functional genomics in fungi. Curr Genet 48, 1–17 (2005). https://doi.org/10.1007/s00294-005-0578-0

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