Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Transformation of Coprinus congregatus with a Linearized Plasmid Vector to Phosphinothricin Resistance

Coprinus congregatus에서 선형으로 전환한 plasmid DNA를 사용하여 phosphinothricin 저항성에 대한 형질전환

  • Leem, Young-Eun (Microbial Phyiology lab, Division of Biological Sciences, Kangwon National University, Research Center for Molecular Microbiology, Seoul National University) ;
  • Kim, Soon-ja (Microbial Phyiology lab, Division of Biological Sciences, Kangwon National University, Research Center for Molecular Microbiology, Seoul National University) ;
  • Choi, Hyoung-Tae (Microbial Phyiology lab, Division of Biological Sciences, Kangwon National University, Research Center for Molecular Microbiology, Seoul National University)
  • 임영은 (강원대학교 생명과학부 미생물학전공, 서울대학교 분자미생물학연구센터) ;
  • 김순자 (강원대학교 생명과학부 미생물학전공, 서울대학교 분자미생물학연구센터) ;
  • 최형태 (강원대학교 생명과학부 미생물학전공, 서울대학교 분자미생물학연구센터)
  • Received : 1997.12.04
  • Accepted : 1997.12.29
  • Published : 1997.12.30
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Abstract

Transformation of Coprinus congregatus with a linearized plasmid has been successfully carried out using phosphinothricin resistance gene as a dominant selectable marker. The transforming frequency was about 500 transformants per microgram of DNA using the protoplast-$CaCl_2$ method. The transforming vector pBARGEM 7-1 which had the phosphinothricin resistance gene was detected in the restriction enzyme fragments of chromosomal DNA from a transformant by Southern hybridization.

Coprinus congregatus에서 laccase를 과량생성하는 변이주를 대상으로 phosphinothricin 저항성을 선택표지로 사용하여 형질전환을 수행하였다. 항생물질에 대한 저항성을 부여하는 plasmid DNA(pBARGEM 7-1)를 제한효소로 가수분해하여 원형질체에 형질전환을 수행한 결과 plasmid DNA $1{\mu}g$ 당 약 500여개의 형질전환체를 얻었다. 도입된 plasmid는 형질전환체의 chromosomal DNA에 삽입되어 있음을 Southern blot으로 확인하였다.

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Acknowledgement

Supported by : 과학재단