효소면역측정법을 이용한 두부 중의 유전자 재조합 대두단백질 분석

Quantification of Genetically Modified Soy Proteins in Fresh Soybean Curd by Antigen-coated Plate ELISA

  • 정미현 (건국대학교 응용생물화학과) ;
  • 배형기 (건국대학교 응용생물화학과) ;
  • 김경미 (건국대학교 응용생물화학과) ;
  • 장인숙 (건국대학교 응용생물화학과) ;
  • 고은정 (건국대학교 응용생물화학과) ;
  • 배동호 (건국대학교 응용생물화학과)
  • Jung, Mee-Hyun (Department of Applied Biology and Chemistry, KonKuk University) ;
  • Bae, Hyung-Ki (Department of Applied Biology and Chemistry, KonKuk University) ;
  • Kim, Kyung-Mi (Department of Applied Biology and Chemistry, KonKuk University) ;
  • Jang, In-Suk (Department of Applied Biology and Chemistry, KonKuk University) ;
  • Ko, Eun-Jung (Department of Applied Biology and Chemistry, KonKuk University) ;
  • Bae, Dong-Ho (Department of Applied Biology and Chemistry, KonKuk University)
  • 발행 : 2004.10.31

초록

본 연구에서는 유전자 재조합 되지 않은(non-GM) 대두와 유전자 재조합된(GM) 대두가 혼입되어 제조된 두부에서 효소 면역 측정법을 이용하여 non-GM 대두의 혼입량을 추정하고자 하였다. 두부의 SDS-PAGE 실행 결과 non-GM 두부에서만 나타나는 특이 단백질 non-GM 113kDa 밴드와 non-GM과 GM 두부에서 모두 나타나는 non-GM 24kDa 밴드를 선별하고 이들을 토끼에 면역하여 항체생성 여부를 ELISA한 결과 non-GM 113kDa과 non-GM 24kDa 단백질 모두 항체가 형성됨을 확인하였고 $10^{-1}-10^{-6}$의 단백질 희석배수에서 두부를 이들 항체에 대하여 ELISA함으로써 원료대두의 GM여부를 확인할 수 있었다. 이들 중, 보다 감도가 높았던 non-GM 113kDa 단백질을 $10^{-7}-10^{-6}$의 배수로 희석하여 ELISA 흡광도와 non-GM 단백질의 관계를 나타내는 표준곡선을 작성하였고, 임의로 non-GM 대두와 GM 대두를 혼합하여 제조한 두부의 ELISA 흡광도를 이 표준곡선과 비교하여 non-GM 원료와 GM 원료 작물의 혼입율을 측정한 결과, 높은 정확도를 보였다.

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