Comparative Study of DNA Fragment According to Steps of Genetically Modified Soybean Processed Food

대두 가공식품의 공정단계별 유전자재조합체(GMO) 단편의 검출확인 및 비교 연구

  • Published : 2003.12.01

Abstract

To discriminate between the genetically modified soybean processed foods which were Korean traditional foods as Beansprout, doenjang, Beancurd (Tofu) and the unmodified one. we had analyzed comparatively that the loss degree of inner DNA about denaturalization factors in process step as heat or pressure and decision of suitable PCR primer by size. As a result of having compared about ${\beta}$-actin was 160bp, 335 promotor was 130 bp and NOS terminator was 132 bp effective. As a result of having checked a loss degree of a gene, as for the bean curd, DNA was mostly preserved well, and the loss of DNA along the processing process was hardly observed by a processing process. Most DNA of beansprout have moved to trunk after germination stage, and the appropriate analysis part was judged as the trunk. And the doenjang showed a detection difference of DNA by an operation of an enzye among self-life periods. Besides, after 50days, insertion gene was destoryed entrely so that detection was not possible.

두부, 콩나물, 된장에서 유전자제조합 대두의 혼입여부를 판별하기 위해 가장 적합한 PCR 프라이머의 선택과 생산 공정에서 물리 ${\cdot}$ 화학적인 변성을 재조합 DNA의 손실정도를 공정단계별로 비교 분석하였다. 내재유전자인 ${\beta}$-actin은 600, 495, 250, 160dp을 비교한 결과 160 bp에서 가장 광범위하게 검출되었으며, 35S promotor는 130bp, NOS terminator 132 bp의 작은 사이즈 프라이머가 유효하였다. 가공공정별로 유전자의 손실정도를 파악한 결과 두부는 대부분 DNA가 잘 보존되어 가공공정에 따른 DNA의 손실은 거의 곤찰되지 않았다. 콩나물은 대부분의 DNA가 발아 직후 줄기로 이동하여 적절한 분석부위는 줄기로 판단되었으며, 된장은 효소의 작용에 듸하여 유통기간내에 DNA의 검출차이를 보였다. 20일 후 삽입유전자는 소실되었고 특히 50일 이후에는 대부분의 내재유전자 뿐만 아니라 분해되어 검출이 어려운 것으로 판단되었다. 가공처리조건인 열에 의한 DNA의 변성은 $100^{\circ}C$에서 40분간 가열하여도 DNA의 손실되지 않고 보존됨을 알 수 있었다.

Keywords

References

  1. Gachet, E., Martin, G.G., Vigneau, F. and Meyer, G.: Detection of genetically modified organisms (GMOs) by PCR: a brief review of methodologies available, Trends in Food Science & Technology, 9, 380-388 (1999) https://doi.org/10.1016/S0924-2244(99)00002-3
  2. ILSI Europe Report Series, Method Development in Relation to Regulatory Requirements for the Detection of GMOs in the food chain, ILSI Europe, 2000
  3. MacCormick, C.A, Griffin, H.G., Underwood, H.M. and Gasson, M.J.: Common DNA sequences with potential for detection of genetically manipulated organisms in food, Journal of Applied Microbiology, 84, 969-980 (1998) https://doi.org/10.1046/j.1365-2672.1998.00429.x
  4. Schreiber, G.A.: Challenges for methods to detect genetically modified DNA in foods, Food Control, 10, 351-352 (1999) https://doi.org/10.1016/S0956-7135(99)00073-0
  5. Hubner, P., Studer, E. & Luthy, J.: Quantitative competitive PCR for the detection of genetically modified organisms in food, Food Control, 10, 353-358 (1999) https://doi.org/10.1016/S0956-7135(99)00074-2
  6. Luthy, J.: Detection strategies for food authenticity and genetically modified foods, Food Control, 10, 359-361 (1999) https://doi.org/10.1016/S0956-7135(99)00075-4
  7. Stave, J.W.: Detection of new of modified proteins in novel foods derived from GMO-future needs, Food Control, 10, 367-374, 1999 https://doi.org/10.1016/S0956-7135(99)00077-8
  8. Van Duijn, G., van Biert, R., Bleeker-Marcelis, H., Peppelman, H. & Hessing, M.: Detection methods for genetically modified crops, Food Control, 10, 375-378 (1999) https://doi.org/10.1016/S0956-7135(99)00078-X
  9. Wurz, A., Bluth, A., Zeitz, P., Pfeifer, C. & Willmund, R.: Quantitative analysis of genetically modified organisms (GMO) in processed food by PCR-based methods, Food Control, 10, 385-389 (1999) https://doi.org/10.1016/S0956-7135(99)00080-8
  10. Meyer, R.: Development and application of DNA analytical methods for the detetction of GMOs in food, Food Control, 10, 391-399 (1999) https://doi.org/10.1016/S0956-7135(99)00081-X
  11. Andreas Zimmermann, Jurg Lithy, Urs Pauli, Quantitative and qualitative evaluation of nine different extration methods for nucleic acids on soya bean food samples, Z Lebensm Unters Forsch A, 207, 81-90 (1998) https://doi.org/10.1007/s002170050299
  12. 藤波傳子, 山口敏利, 毛利光之: 된장 숙성 과정 중에 대두유래 DNA의 消辰-재조합DNA검증-, 된장의 과학과 기술, 48(11), 399-402, 2000