title: T-DNA transfer and integration in the ectomycorrhizal fungus Suillus bovinus using hygromycin B as a selectable marker.
abstract: The T-DNA of Agrobacterium tumefaciens can be transferred to plants, yeasts, fungi and human cells. Using this system, dikaryotic mycelium of the ectomycorrhizal fungus Suillus bovinus was transformed with recombinant hygromycin B phosphotransferase (hph)and enhanced green fluorescent protein (EGFP) genes fused with a heterologous fungal promoter and CaMV35S terminator. Transformation resulted in hygromycin B-resistant clones, which were mitotically stable. Putative transformants were analysed for the presence of hph and EGFP genes by PCR and Southern analysis. The latter analysis proved both multiple- and single-copy integrations of the genes in the S. bovinus genome. A. tumeficiens transformation should make possible the development of tagged mutagenesis and targeted gene disruption technology for S. bovinus.
authors.affiliation: Department of Biosciences, University of Helsinki, Finland. hanif@mappi.helsinki.fi
authors.forename: Mubashir
authors.initials: M
authors.lastname: Hanif
authors.forename: Alejandro Guillermo
authors.initials: AG
authors.lastname: Pardo
authors.forename: Markus
authors.initials: M
authors.lastname: Gorfer
authors.forename: Marjatta
authors.initials: M
authors.lastname: Raudaskoski
mesh_terms: D001483:Base Sequence; D001487:Basidiomycota / Q000235:genetics*; D015139:Blotting, Southern; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics*; D005819:Genetic Markers; D006921:Hygromycin B / Q000494:pharmacology*; D014170:Transformation, Genetic
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D005819:Genetic Markers; C036483:T-DNA; D006921:Hygromycin B