Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Genetic Transformation of Irpex lacterus and Phlebia tremellosa to an Antibiotic Resistance

아교버섯과 기계충버섯의 형질전환

  • Kim, Yun-Jung (Division of Life Sciences, and Research Institute of Life Sciences, Kangwon National University) ;
  • Kim, Myung-Kil (Division of Wood Chemistry and Microbiology, Korea Forest Research Institute) ;
  • Song, Hong-Gyu (Division of Life Sciences, and Research Institute of Life Sciences, Kangwon National University) ;
  • Choi, Hyoung-T. (Division of Life Sciences, and Research Institute of Life Sciences, Kangwon National University)
  • 김윤정 (강원대학교 생명과학부 생명과학연구소) ;
  • 김명길 (국립산림과학원 화학미생물과) ;
  • 송홍규 (강원대학교 생명과학부 생명과학연구소) ;
  • 최형태 (강원대학교 생명과학부 생명과학연구소)
  • Published : 2007.06.30
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Abstract

White-rot fungi which degrade lignin can also degrade diverse recalcitrant compounds such as polymeric dyes, explosives, pesticides, and endocrine disrupting chemicals. Lignin degrading enzymes are involved in the degradation reactions, and introduction of foreign genes into a white-rot fungus is required in order to increase the degrading capacity. Genetic transformation experiment has been carried out in Irpex lacteus and Phlebia tremellosa to an antibiotic resistance. The transformation yields were 50-70 transformants/${\mu}g$ DNA and 15-25 transformants/${\mu}g$ DNA in I. lacteus and P. tremellosa, respectively. The stable replication of the plasmid was confirmed by PCR using the plasmid-specific primers, and many mutants were generated during this integration in both fungi.

국내에서 분리한 백색부후균 기계충버섯(Irpex lacteus)과 아교버섯(Phlebia tremellosa)을 대상으로 유전자 도입을 위한 형질전환 방법을 확립하였다. 아교버섯의 정우 원형질체 생성 및 재생 방법을 이용하여 이핵체(dikaryon)로부터 일핵체(monokaryon: Pt05-2)를 얻어 실험에 사용하였다. 형질전환체의 선발을 위한 선택표지는 glutamine synthetase의 inhibitor인 phosphinothricin에 대하여 저항성을 부여하는 유전자(bar)를 사용하였고, 이 유전자를 가진 형질전환용 벡터, pBARGEM7-1 ($5\;{\mu}g$)과 제한효소 EcoRI (30 u)를 동시에 원형질체에 처리하는 제한효소매개 삽입방법을 사용하였다. 기계충버섯($5{\times}10^{7}\;cells$)및 아교버섯($2.5{\times}10^{7}\;cells$)의 원형질체를 대상으로 형질전환을 수행한 결과 벡터 $1\;{\mu}g$당 각각 형질전환체 50-70개 및 15-25개의 수율을 보였으며 형질전환용 벡터가 각 형질전환체에 안정되게 존재함을 확인하였다.

Keywords

References

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