Transformation of Pleurotus florida with Neurospora pyr 4 Gene

Neurospora pyr 4 유전자를 이용한 사철 느타리버섯의 형질전환

  • Byun, Myung-Ok (Applied Mycology and Mushroom Division, Agricultural Sciences Institute, R.D.A.) ;
  • Yoo, Young-Bok (Applied Mycology and Mushroom Division, Agricultural Sciences Institute, R.D.A.) ;
  • You, Chang-Hyun (Applied Mycology and Mushroom Division, Agricultural Sciences Institute, R.D.A.) ;
  • Cha, Dong-Yeul (Applied Mycology and Mushroom Division, Agricultural Sciences Institute, R.D.A.) ;
  • Cho, Moo-Je (Department of Biochemistry, Gyeongsang National University)
  • 변명옥 (농촌진흥청 농업기술연구소 균용과) ;
  • 유영복 (농촌진흥청 농업기술연구소 균용과) ;
  • 유창현 (농촌진흥청 농업기술연구소 균용과) ;
  • 차동열 (농촌진흥청 농업기술연구소 균용과) ;
  • 조무제 (경상대학교 생화학과)
  • Published : 1989.12.01

Abstract

Transformation of an auxotrophic requirement for uracil in Pleurotus florida P101 has been achieved using chimeric vector containing Aspergillus nidulans ans 1, and Neurospora crassa pyr 4 DNA. Protoplasts of $Ura^-$strains of P. florida were incubated with plasmid pDJB3 containing the cloned pyr 4 gene in the presence of polyethylene glycol and $CaCl_2$. Transformants could grow on MMM showing mitotical stability. Southern hybridization analysis of DNA isolated from transformants showed that the Neurospora pyr 4 gene and vector sequence might be integrated into the P. florida chromosomes. As the transformants were monokaryon, each transformant was mated with the other monokaryon. Fruitbody shape of untransformant was eroded type but those of transformants were eroded type, funnel type, plane type and ungrowing cap type.

사철 느타리 버섯 Uracil 요구성 균주를 Aspergillus nidulans ans 1과 Neurospora crassa pyr 4 유전자를 지닌 백터를 이용하여 형질전환시켰다. 사철 느타리버섯 ura 요구성 균주의 원형질체를 pyr 4 유전자를 지닌 백터 pDJB3과 혼합 후 PEG와 $CaCl_2$를 처리하였다. 형질전환 균주는 버섯 최소배지에서 안정되게 생육하였고 southern hybridization 결과도 백터 DNA가 사철 느타리 염색체에 삽입되었다. 형질전환 균주는 단핵성이므로 다른 단핵 균사와 균사 융합 후 자실체 형태, 포자형성 등을 조사하였다. 형질전환이 안된 균주는 톱니형 자실체를 형성한데 비해 형질전환된 균주는 톱니형, 깔대기형, 우산형, 갓 미발육형의 자실체를 형성하였다.

Keywords