• Korean Journal of Microbiology
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• v.50 no.4
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• pp.372-375
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• 2014

This experiment was undertaken to investigate proper conditions for protoplast isolation and genetic transformation of the white rot fungi, Polyporus brumalis. The protoplasts were formed from mycelia at a frequency of $1{\times}10^7/ml$ with 0.5% Usukizyme. The transformation vector (pHYgpt) was constructed using hygromycin resistance gene (hph) for the selectable maker. The yield was 100-160 transformants/${\mu}g$ DNA in a transformation mediated by 40% polyethylene glycol solution with aurintricarboxylic acid, heparin and supermidine. The genomic integration of the pHYgpt was confirmed by hph-specific PCR and the expected amplified band appeared only in the transformants. These results could be an efficient tool in gene engineering of the genus polyporus.