Horticultural Science & Technology (원예과학기술지)

Korean Society of Horticultural Science (한국원예학회)
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Identification of Introduced Gene and Its Expression and Gene Stability Assessment for Event Selection of Genetically Modified Plant toward Approval: Cucumber Mosaic Virus Resistant Hot Pepper

상업용 유전자 변형작물 이벤트 선발을 위한 도입유전자 확인, 발현 및 세대간 안정성 평가 : 오이모자이크바이러스 저항성 GM 고추

  • Kang, Seung-Won (BET Institute, Chung-Ang University) ;
  • Han, Bal-Kum (Department of Integrative Plant Science, Chung-Ang University) ;
  • Lee, Tae-Ho (Department of Integrative Plant Science, Chung-Ang University) ;
  • Kim, Eun-Ji (Department of Integrative Plant Science, Chung-Ang University) ;
  • Lee, Gung-Pyo (Department of Integrative Plant Science, Chung-Ang University)
  • 강승원 (중앙대학교 생명환경연구원) ;
  • 한밝음 (중앙대학교 생명자원공학부 식물시스템과학전공) ;
  • 이태호 (중앙대학교 생명자원공학부 식물시스템과학전공) ;
  • 김은지 (중앙대학교 생명자원공학부 식물시스템과학전공) ;
  • 이긍표 (중앙대학교 생명자원공학부 식물시스템과학전공)
  • Received : 2012.01.02
  • Accepted : 2012.01.31
  • Published : 2012.04.30
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Abstract

For the development of genetically modified plants, it is important to verify various factors which potentially affect the risk assessment as well as to establish an experimental program to produce scientific and reliable data. However, it is a time and cost consuming process to develop GM plants as well as to prepare scientific and convincible data for government's approval. Therefore, using the transgenic hot pepper tolerant to a new CMV pathotype, we attempted to suggest few methodological procedures, such as probe saturation for southern blot analysis and RT-PCR and ELISA for expression analysis, for identification and stability evaluation of inserted gene in genetically modified plant which are required for submission for approval. Ten partially overlapped probes covering full length of inserted gene were produced. We could identify that the inserted gene was stacked as a single copy as well as no partial element existed. Also, we could identify the stability of the inserted gene stacked in hot pepper using probe saturation. In the expression analysis with RT-PCR and ELISA, we also could provide the stable expression of transcript and proteins in leaves and placenta and pericarp of fruits of the CMV-resistant hot pepper.

형질전환 작물 개발시, 위해성 평가에 영향을 미치게 될 요인들을 파악하고, 이에 대한 과학적 근거의 마련을 위한 실험 자료 생산계획의 확립과 실험을 진행하는 것이 매우 중요하다. 또한 유전자변형작물을 개발한 후 정부의 승인을 위한 과학적이고 신뢰할 만한 자료를 축적하기 위해서는 오랜 기간과 많은 자본이 투입된다. 이에 따라, 본 연구에서는 CMV 저항성 유전자변형 고추에 대한 평가분석 연구를 통해 유전자변형작물의 환경위해성 평가를 수행하는데 있어 도입유전자 확인을 위한 평가서 작성을 위해 요구되는 서던 분석을 위한 탐침 포화, 도입유전자의 발현 해석을 위한 RT-PCR, ELISA 실험에 관한 사례를 제시하고자 하였다. 서던 분석을 위해 삽입 유전자 전체 서열의 확인이 가능한 10개의 부분적으로 겹치는 탐침을 제작하였다. 서던 분석 결과, 고추에 도입된 외래유전자가 단일사본으로 존재하여, 게놈전체에서 도입된 위치 이외에 상동성을 갖는 다른 조각 부위가 없음을 확인하였으며, 도입유전자 전체의 안정성을 확실히 입증할 수 있었다. 그와 더불어, 역전사 중합효소 연쇄반응 및 효소면역법 실험을 통하여, CMV 저항성 고추의 잎, 과실 등과 같은 주요 기관에서의 삽입 유전자에 대한 안정적인 발현을 확인 할 수 있었다.

Keywords

References

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