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

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Author: Michielse, C.B. · Hooykaas, P.J.J. · Hondel, C.A.M.J.J. van den · Ram, A.F.J.
Type:article
Date:2005
Institution: TNO Kwaliteit van Leven
Source:Current Genetics, 1, 48, 1-17
Identifier: 238591
doi: doi:10.1007/s00294-005-0578-0
Keywords: Biology · Analytical research · Agrobacterium tumefaciens · Fungal transformation · Insertional mutagenesis · Targeted mutagenesis · bacterial DNA · DNA integration · DNA synthesis · DNA transfer · functional genomics · fungal genetics · fungal strain · gene expression regulation · gene targeting · genetic transformation · molecular genetics · nonhuman · priority journal · review · Rhizobium · Rhizobium radiobacter · Bacterial Proteins · Base Sequence · DNA, Fungal · Genome, Fungal · Genomics · Mutagenesis, Insertional · Mutagenesis, Site-Directed · Plasmids · Rhizobium · Saccharomyces cerevisiae · Agrobacterium · Agrobacterium tumefaciens · Fungi

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. © Springer-Verlag 2005.