Transformation of Pleurotus sajor-caju by Complementation of PABA Requiring Mutant

여름 느타리버섯에서 PABA 변이주의 Complementation에 의한 형질전환

  • Byun, Myung-Ok (Department of Molecular Genetics, National Institute of Agricultural Science and Technology) ;
  • Chung, Jong-Chun (Department of Molecular Genetics, National Institute of Agricultural Science and Technology) ;
  • You, Chang-Hyun (Department of Molecular Genetics, National Institute of Agricultural Science and Technology) ;
  • Cha, Dong-Yeul (Department of Molecular Genetics, National Institute of Agricultural Science and Technology) ;
  • Lee, Du-Hyung (Department of Env. Hort., Seoul City University)
  • 변명옥 (농업과학기술원 분자유전과) ;
  • 정종천 (농업과학기술원 응용미생물과) ;
  • 유창현 (농업과학기술원 응용미생물과) ;
  • 차동열 (농업과학기술원 응용미생물과) ;
  • 이두형 (서울시립대학교 환경원예학과)
  • Published : 1997.09.30

Abstract

A PABA auxotroph of Pleurotus sajor-caju were transformed to prototrophy by using a plasmid containing pab 1 gene from Coprinus. The efficiencies of transformation of Pleurotus sajor-caju was five transformants per ${\mu}g$ of plasmid DNA. Southern blot analysis of DNA extracted from transformants demonstrated that plasmid DNA was integrated into the chromosomal DNA in multiple tandem copies. Progenies of heterokaryons between transformants of PABA and other auxotropic strains produced pab-progeny, which indicated that integration occurred at a site(s) other than the resident pab biosynthetic gene.

여름느타리버섯의 단포자에 UV를 처리하여 pab 요구성 영양요구주를 작성하였으며 이 균의 균사에서 원형질체를 분리한 후 Coprinus pab 1유전자를 함유하는 plasmid를 이용하여 prototrophy로 형질전환 하였다. 형질전환율은 ${\mu}g$의 DNA 당 5개의 형질전환주를 얻을 수 있었다. 형질전환주는 Southern 분석결과 염색체 DNA 속으로 integration된 것으로 확인되었으며 영양생장과 생식 생장시 모두 안정하게 유지되었다. 형질전환주와 화합성인 다른 영양요구주와 교배후 자실체의 포자를 분리하여 유전분석 결과 pab 생합성 유전자 보다는 그 유전자 주위에 integration이 일어난 것으로 추정되었다.

Keywords

References

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