Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Detection of Genetically Modified Soybean in Tofu and Biji using PCR and Immunological Methods

PCR 방법과 면역학적 분석법을 이용한 두부와 비지에서 GM 콩의 검출법

  • Kim, Myo-Young (Department of Food Science and Biotechnology and Institute of Life Sciences & Resources, Kyung Hee University) ;
  • Kim, Jae-Hwan (Department of Food Science and Biotechnology and Institute of Life Sciences & Resources, Kyung Hee University) ;
  • Kim, Hae-Yeong (Department of Food Science and Biotechnology and Institute of Life Sciences & Resources, Kyung Hee University)
  • 김묘영 (경희대학교 식품생명공학과 및 생명자원과학연구소) ;
  • 김재환 (경희대학교 식품생명공학과 및 생명자원과학연구소) ;
  • 김해영 (경희대학교 식품생명공학과 및 생명자원과학연구소)
  • Published : 2005.03.31
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Abstract

To monitor GM soybean in soybean processed foods, tofu and biji, we prepared tofu and biji containing 0%, 1%, 3%, 5% and 100% GM soybean, respectively. We examined epsps gene inserted in soybean by PCR and EPSPS protein expressed in soybean using western blotting and lateral flow strip test to compare the sensitivity of these methods. A PCR product of 123 bp inserted in GM soybean was detected in all tofu and biji containing 1%, 3%, 5% and 100% GM soybean with the exception of 0% samples; however, the size of 600 bp inserted in GM soybean was only detected in tofu containing 100% soybean and in biji containing 5% and 100% soybean. In the protein level, GM soybean product was only detected in tofu and biji containing 100% GM soybean by western blotting. In addition, only biji containing 100% GM soybean was detected by lateral flow strip test. We concluded that in order to detect GM soybean efficiently in processed food, the PCR method is more sensitive than immunological methods. With the PCR method, small size product with approximately 100 bp in PCR product is sensitive to detect GM soybean in processed foods.

GM 콩으로 제조된 두부와 비지 등 콩 가공제품들의 효율적인 모니터링을 위하여 GM 콩이 0%, 1%, 3%, 5%, 100% 함유된 두부와 비지를 제조하여 이들의 삽입유전자인 5-enolpyruvylshikimate-3-phosphate synthase(epsps)를 검출하기 위한 PCR 방법과 EPSPS 발현 단백질 검출을 위한 western blotting과 lateral flow strip을 사용하여 검출 감도를 비교하였다. PCR 결과 산물의 크기가 큰 600 bp의 증폭에서 두부의 경우에는 100% GM으로 제조된 두부에서만 확인이 가능하였고, 비지에서는 5%, 100%에서 확인이 되었다. 크기가 작은 123 bp가 생성되는 실험에서는 0%를 제외한 모든 두부와 비지에서 검출되었다. Western blot 결과는 100% 두부, 비지에서만 검출이 되었고, lateral flow strip test에서는 100% 비지에서만 검출되었다. 이러한 결과로부터 GM 콩 가공식품 검출은 면역학적 방법보다는 PCR이 더 높은 민감도가 나타내는 것을 확인하였고, 또한 가공식품에서 효율적인 GMO 검출을 위해서는 100 bp 정도의 작은 크기의 PCR 산물을 이용해야 민감한 검출 결과를 보여주는 것을 확인하였다.

Keywords

References

  1. James, C. (2004) Preview: Global status of commercialized biotech/GM crops: ISAAA Briefs No. 32. ISAAA: Ithaca, N.Y
  2. Kim, M. Y., Kim, J. H., Kim, H. J., Park, S. H., Woo, G. J. and Kim, H. Y. (2003) Monitoring of genetically modified soybean and processed foods in Korean market using PCR. J. Korean Soc. Agric. Chem. Biotechnol. 46, 344-347
  3. Kim, J. H. (2003) Studies on the Polymerase Chain Reaction for the Rapid Detection of Genetically Modified Foods. MS Thesis. Kyunghee University
  4. Auer, C. A. (2003) Tracking genes from seed to supermarket: techniques and trends. Trends Plant Sci. 8, 591-597 https://doi.org/10.1016/j.tplants.2003.10.010
  5. Ahmed, F. E. (2002) Detection of genetically modified organisms in foods. Trends Biotech. 5, 215-223
  6. Ministry of Agriculture and Forestry. (2000) Regulation concerning the compulsory labeling of genetically modified agricultural organisms. Ministry of Agriculture and Forestry regulation No. 2000-31
  7. Korea Food and Drug Administration. (2001) Regulation concerning the compulsory labeling of foodstuffs and food ingredients produced from genetically modified organisms. Korea Food and Drug Administration regulation No. 2001-43
  8. Kim, H. J., Park, S. H. and Kim, H. Y. (2001) Study for detection of glyphosate tolerant soybean using PCR. Korean J. Food Sci. Technol. 33, 521-524
  9. Kwak, B. Y., Ko, S. H., Park, C. W., Son, D. Y. and Shon, D. H. (2003) Development of enzyme-linked immunosorbent assay for glyphosate-tolerant soybeans. Korean J. Food Sci. Technol. 35, 366-372
  10. Kim, Y. M., Sohn, S. H., Jeong, S. I., Yoon, M. S., Kim, T. S. and Park, Y. H. (2002) Detection methods for genetically modified soybeans. J. Korean Soc. Agric. Chem. Biotechnol. 45, 185-189
  11. Padgette, S. R., Kolacz, K. H., Delannay, X., Re, D. B., Lavallee, B. J., Tinius, C. N., Rhodes, W. K., Otero, Y. I. and Barry, G. F. (1995) Development, identification, and characterization of a glyphosate-tolerant soybean line. Crop Sci. 35, 1451-1461 https://doi.org/10.2135/cropsci1995.0011183X003500050032x
  12. Barry, G. F., Kishore, G. M., Padgette, S. R. and Stallings, W. C. (1997) Glyphosate-tolerant 5-enolpyruvyl-3-phosphate. U.S. Patent 5,633,435
  13. Proudhon, D., Wei, J., Briat, J. and Theil, E. C. (1996) Ferritin gene organization: differences between plants and animal suggest possible kingdom-specific selective constraints. J. Mol. Evol. 42, 325-336 https://doi.org/10.1007/BF02337543