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Protoplast Isolation and Genetic Transformation of Polyporus brumalis

겨울우산버섯의 원형질체 분리와 유전자 형질전환

  • Ryu, Sun-Hwa (Division of Wood Chemistry & Microbiology, Korea Forest Research Institute (KFRI)) ;
  • Kim, Myung-Kil (Division of Wood Chemistry & Microbiology, Korea Forest Research Institute (KFRI))
  • 유선화 (국립산림과학원 임산공학부 화학미생물과) ;
  • 김명길 (국립산림과학원 임산공학부 화학미생물과)
  • Received : 2014.10.27
  • Accepted : 2014.11.22
  • Published : 2014.12.31

Abstract

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.

Keywords

Polyporus brumalis;protoplast;pYHgpt;transformation;white rot fungi

Acknowledgement

Supported by : 국립산림과학원

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