T-DNA transfer and integration in the ectomycorrhizal fungus Suillus bovinus using hygromycin B as a selectable marker.
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.
Hanif
Mubashir
M
Department of Biosciences, University of Helsinki, Finland. hanif@mappi.helsinki.fi
Pardo
Alejandro Guillermo
AG
Gorfer
Markus
M
Raudaskoski
Marjatta
M
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
D017931:DNA Primers; D004269:DNA, Bacterial; D005819:Genetic Markers; C036483:T-DNA; D006921:Hygromycin B