担子菌へのプラスミドDNA導入とその発現についての予備的研究

To attempt the gene transfer to the basidiomycetes such as wood-decaying fungi, the experimental system of transferring plasmid DNA and seeing the expression was investigated. Using two species of Tyromyces palustris and Coriolus versicolor, their protoplasts from mycelia cultivated in SMY liquid culture media were prepared by treatments with the cell wall-lysing enzymes and others. As plasmid DNA, pUC4K having a kanamycin-resistant gene was selected because of obstructing the growth of mycelia. The transfer of plasmid DNA was examined by two ways of polyethylene glycol and electro poration.
The transformants of Tyromyces palustris obtained by electroporation showed obviously a faster growth than the non-transformed cells in kanamycincontaining liquid media. As a result of agarose gel electrophoresis for chromosomal DNA and plasmid DNA extracted from these transformants, the bands could be detected at the same position of plasmid pUC4K. This verified the transformation to Tyromyces palustris by transferring pUC4K.
Keywords: transformation; basidiomycetes; Tyromyces palustris; Plasmid pUC4K; electroporation.