다섯 가지 DNA 추출방법에 의한 옥수수 원료 및 가공시료의 DNA 추출 효율의 비교

Comparison of the Efficiency from Raw and Processed Corns by Five Different DNA Extraction Methods

  • 이훈희 (식품의약품안전청 영양평가팀) ;
  • 송희성 (경희대학교 생명자원과학연구원) ;
  • 김재환 (경희대학교 생명자원과학연구원) ;
  • 이우영 (식품의약품안전청 영양평가팀) ;
  • 이순호 (식품의약품안전청 영양평가팀) ;
  • 박선희 (식품의약품안전청 영양평가팀) ;
  • 박혜경 (식품의약품안전청 영양평가팀) ;
  • 김해영 (경희대학교 생명자원과학연구원)
  • Lee, Hun-Hee (Team of Nutrition Evaluation, Korea Food and Drug Administration) ;
  • Song, Hee-Sung (Institute of Life Sciences & Resources, Kyung Hee University) ;
  • Kim, Jae-Hwan (Institute of Life Sciences & Resources, Kyung Hee University) ;
  • Lee, Woo-Young (Team of Nutrition Evaluation, Korea Food and Drug Administration) ;
  • Lee, Soon-Ho (Team of Nutrition Evaluation, Korea Food and Drug Administration) ;
  • Park, Sun-Hee (Team of Nutrition Evaluation, Korea Food and Drug Administration) ;
  • Park, Hye-Kyung (Team of Nutrition Evaluation, Korea Food and Drug Administration) ;
  • Kim, Hae-Yeong (Institute of Life Sciences & Resources, Kyung Hee University)
  • 발행 : 2005.12.31

초록

본 연구는 일반적으로 사용되는 DNA 추출방법들에 대한 효율을 비교하기 위해 수행하였다. 원료 옥수수 및 이를 가공한 시료들로부터 DNA추출을 하였으며, 추출된 DNA의 형상, 농도 및 순도 측정 그리고 PCR 분석 결과를 비교하였다. 5가지 방법으로 옥수수의 DNA를 추출한 결과, 추출방법에 따른 DNA 형상의 차이가 거의 없는 것을 확인하였으나, 각각의 시료들로부터 추출된 DNA의 양은 시료 g당 $0.25{\mu}g$부터 $234.0{\mu}g$까지 매우 다양하게 나타났다. 5가지 방법으로 옥수수 시료들의 DNA를 추출한 결과, CTAB법과 DNeasy plant Maxi 키트를 이용한 DNA 추출방법이 높은 수율을 보였다.

In this study, the effects of five extraction methods for raw and processed corns were compared with respect to the integrity, yields and quality of DNA extracted from them and the results were assessed by PCR analysis. From the comparison of five extraction methods, DNA integrity showed a similar pattern. Amounts of genomic DNA obtained from the five extraction methods varies from $0.25{\mu}g\;to\;234{\mu}g$ per 1 g sample. The DNA yield extracted with CTAB method and DNeasy Plant Maxi kit is greater than that obtained from other extraction methods. These results would be applicable for the selection of an adequate extraction method for specific samples.

키워드

참고문헌

  1. USDA (2005) World Agricultural Supply Demand Estimates, WASDE-4
  2. Clive, J. (2004) Preview: Global status of commercialized biotech/GM crops. ISAAA briefs No. 32, ISAAA: Ithaca, NY
  3. 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
  4. 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
  5. Bertheau, Y., Diolez, A., Kobilinsky, A. and Magin, K. (2002) Detection methods and performance criteria for genetically modified organisms. J. AOAC. Int. 85, 801-808
  6. Vollenhofer, S., Burg, K., Schmidt, J. and Kroath, H. (1999) Genetically modified organisms in food-screening and specific detection by polymerase chain reaction. J. Agric. Food Chem. 47, 5038-5043 https://doi.org/10.1021/jf990353l
  7. 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
  8. Hur, M. S., Kim, J. H., Park, S. H., Woo, G. J. and Kim, H. Y. (2003) Detection of Genetically Modified Maize Safety-approved in Korea using PCR. Korean J. Food Sci. Technol. 35, 1033-1038
  9. Lee, S. H., Park, Y. H., Kim, J. K., Park, K. W. and Kim, Y. M. (2004) Qualitative PCR Method for Detection of Genetically Modified Maize Lines NK603 and TC1507. Agric. Chem. Biotechnol. 47, 185-188
  10. Rogan, G. J., Dudin, Y. A., Lee, T. C., Magin, K. M., Astwood, J. D., Bhakta, N. S., Leach, J. N., Sanders, P. R. and Fuchs, R. L. (1999) Immunodiagnostic methods for detection of 5-enolpyruvyl-shikimate-3-phosphate synthase in roundup ready soybeans. Food Control 10, 407-414 https://doi.org/10.1016/S0956-7135(99)00083-3
  11. Trifa, T. and Zhang, D. (2004) DNA content in Embryo an Endosperm of Maize Kernel (Zea mays L.): Impact on GMO Quantification. J. Agric. Food Chem. 52, 1044-1048 https://doi.org/10.1021/jf034574+
  12. Peano, C., Samson, M. C., Palmieri, L., Gulli, M. and Marmiroli, N. (2004) Qualitative and Quantitative Evaluation of the Genomic DNA Extracted from GMO and Non-GMO Foodstuffs with Four Different Extraction Methods. J. Agric. Food Chem. 52, 6962-6968 https://doi.org/10.1021/jf040008i
  13. Lee, C. H., Kim, D. C., Chun, J. H., Kim, C. J., Kim, J. B., Kim, J. D. and Son, J. C. (1987) Food Extrusion Technology. Yu-Lim Publishing Co., Korea
  14. Ham, C., Knight, M., Ramakrishnan, A., Guan, H., Keeling, P. L. and Wasserman B. P. (1998) Isolation and Characterization of the zSSIIa and zSSIIb Starch synthase cDNA clones from Maize Endosperm. Plant Mol. Biol, 37, 639-649 https://doi.org/10.1023/A:1006079009072
  15. Meyer, R. (1999) Development and Application of DNA Analytical Methods for the Detection of GMOs in Food. Food Control 10, 391-399 https://doi.org/10.1016/S0956-7135(99)00081-X
  16. Kim, M. Y., Kim J. H., 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