Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Development of Detection Method of Unapproved Genetically Modified Potato (EH92-527-1) in Korea using Duplex Polymerase Chain Reaction

Duplex PCR을 이용한 국내 미승인 유전자변형 감자(EH92-527-1)의 검사법 개발

  • Yoo, Myung-Ryul (Institute of Life Sciences and Resources and Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Kim, Jae-Hwan (Institute of Life Sciences and Resources and Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Yea, Mi-Chi (Department of plant Quarantine, Animal, Plant and Fisheries Quarantine and Inspection Agency) ;
  • Kim, Hae-Yeong (Institute of Life Sciences and Resources and Department of Food Science and Biotechnology, Kyung Hee University)
  • 유명렬 (경희대학교 생명자원과학연구원 및 식품생명공학과) ;
  • 김재환 (경희대학교 생명자원과학연구원 및 식품생명공학과) ;
  • 예미지 (농림수산검역검사본부 식물검역부 위험관리과) ;
  • 김해영 (경희대학교 생명자원과학연구원 및 식품생명공학과)
  • Received : 2012.12.03
  • Accepted : 2013.01.02
  • Published : 2013.04.30
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Abstract

A duplex polymerase chain reaction (PCR) method was developed to detect unapproved genetically modified (GM) potato (EH92-527-1) in Korea. The UDP-glucose pyrophosphorylase (UGP) gene was selected as an endogenous reference gene for potato and used to validate the specificity for 14 different crops. The primer pair EH92-F/R was designed to amplify the junction sequence between the genome and transgenic region introduced in GM potato. Its specificity was also validated using several different GM events. The detection limit of the duplex PCR method is approximately 0.05%. This duplex PCR method could be useful for monitoring cultivation of unauthorized GM potato in Korea.

우리나라에서 미승인 품목인 유전자변형 감자 EH92-527-1를 검출하기 위한 duplex PCR 검사법이 개발되었다. 감자의 내재유전자로 UDP-glucose pyrophosphorylase (UGP)가 선별되었고, 14개 다른 작물을 이용하여 특이성이 확인되었다. 유전자변형 감자에 삽입된 T-DNA 영역과 감자 게놈 사이의 연결 부위를 증폭하도록 프라이머 EH92-F/R 쌍이 제작되었고, 몇 개의 다른 유전자 변형 작물을 이용하여 특이성이 확인되었다. 서론에서 언급한 바와 같이 BASF사에서 각 개발된 유전자변형 감자 EH92-527-1과 BPS-A1020-5가 GBSS 유전자를 동일하게 포함하고 있으나 본 연구에서 개발한 검사법은 event-specific primers를 이용하였기 때문에 유전자변형 감자 EH92-527-1에만 특이성을 나타낸다. 이와 같이 개발된 duplex PCR 검사법의 검정한계치는 약 0.05%이다. 이러한 duplex PCR 검사법이 우리나라에 미승인 유전자변형 감자의 모니터링에 유용하게 사용될 것으로 판단한다.

Keywords

References

  1. International Service for the Acquisition of Agri-biotech Applications (ISAAA). GM Approval Database. Available from: http:// www.isaaa.org. Accessed Nov. 30, 2012.
  2. Korea Biosafety Clearing House. Status of risk assessment of GMO in Korea. Available from: http://www.biosafety.or.kr. Accessed Nov. 30, 2012.
  3. Center for Environmental Risk Assessment (CERA), ILSI Research Foundation. GM Crop Database. Available from: http:// www.cera-gmc.org. Accessed Nov. 30, 2012.
  4. KREI. Outlook of the Agricultural Economy. Korea Rural Economic Institute, Seoul, Korea. pp. 82-84 (2011)
  5. EFSA. Opinion of the scientific panel on genetically modified organisms on a request from the commission related to the notification (reference C/SE/96/3501) for the placing on the market of genetically modified potato EH92-527-1 with altered starch composition, for cultivation and production of starch, under part C of directive 2001/18/EC from BASF Plant Science. The EFSA Journal 323: 1-20 (2006)
  6. Kim JH, Ahn JH, Song HS, Kim KH, Kim DH, Kim HY. Qualitative PCR detection of vitamin E-enriched GM perilla. J. Korean Soc. Appl. Biol. Chem. 49: 192-195 (2006)
  7. Oguchi T, Onishi M, Chikagawa Y, Minegishi Y, Kodama T, Akiyama H, Ohno Y, Futo S, Hino A, Furui S, Kitta K. Development of event-specific quantitation method for GA21 maize, which is a GM event without CaMV35S promoter. J Food Hyg. Soc. Jpn. 49: 16-22 (2008) https://doi.org/10.3358/shokueishi.49.16
  8. Yang L, Xu S, Pan A, Yin C, Zhang K, Wang Z, Zhou Z, Zhang D. Event specific qualitative and quantitative polymerase chain reaction detection of genetically modified MON863 maize based on the 5'-transgene integration sequence. J. Agric. Food Chem. 53: 9313-9318 (2005)
  9. Holck A, Vaitilingom M, Didierjean L, Rudi K. 5'-Nuclease PCR for quantitative event-specific detection of the genetically modified Mon810 MaisGard maize. Eur. Food Res. Technol. 214: 449- 453 (2002) https://doi.org/10.1007/s00217-001-0473-y
  10. Kim SA, Lee MK, Yoo SJ, Kim JH, Lee H, Park KS, Jeong J, Kim HY. Detection of GM papaya event 55-1 in fresh and processed papaya using duplex PCR. J. Korean Soc. Appl. Biol. Chem. 53: 237-242 (2010) https://doi.org/10.3839/jksabc.2010.037
  11. Mano J, Furui S, Takashima K, Koiwa T, Futo S, Minegishi Y, Akiyama H, Teshima R, Kurashima T, Takabatake R, Kitta K. Development and validation of event-specific quantitative PCR method for genetically modified maize MIR604. J. Food Hyg. Soc. Jpn. 53: 166-171 (2012) https://doi.org/10.3358/shokueishi.53.166
  12. Li X, Pan L, Li J, Zhang Q, Zhang S, Lv R, Yang L. Establishment and application of event-specific polymerase chain reaction methods for two genetically modified soybean events, A2704-12 and A5547-127. J. Agric. Food Chem. 59: 13188-13194 (2011) https://doi.org/10.1021/jf202806w
  13. Taverniers I, Windels P, Vatilingom M, Milcamps A, Van Bockstaele E, Van den Eede G, De Loose M. Event-specific plasmid standards and real-time PCR methods for transgenic Bt11, Bt176, and GA21 maize and transgenic GT73 canola. J. Agric. Food Chem. 53: 3041-3052 (2005) https://doi.org/10.1021/jf0483467
  14. Rho JK, Lee T, Jung SI, Kim TS, Park YH, Kim YM. Qualitative and quantitative PCR methods for detection of three lines of genetically modified potatoes. J. Agric. Food Chem. 52: 3269- 3274 (2004) https://doi.org/10.1021/jf0499020
  15. Watanabe T, Kuribara H, Mishima T, Kikuchi H, Kodama T, Futo S, Kasama K, Toyota A, Nouno M, Saita A, Takahashi K, Hino A, Akiyama H, Maitani T, Kubo M. New qualitative detection method of genetically modified potatoes. Biol. Pharm. Bull. 27: 1333-1339 (2004) https://doi.org/10.1248/bpb.27.1333
  16. Broothaerts W, Corbisier P, Emons H, Emterborg H, Linsinger TPJ, Trapmann S. Development of a certified reference material for genetically modified potato with altered starch composition. J. Agric. Food Chem. 55: 4728-4734 (2007) https://doi.org/10.1021/jf0701584
  17. BASF Plant Science GmbH. Event-specific method for the quantification of amylopectin potato event EH92-527-1 using real-time PCR, Ludwigshafen, Germany. pp. 1-12 (2009)
  18. Hofvander P. Structure and DNA sequence of insert and flanking genomic region of potato event EH92-527-1. BASF Plant Science GmbH, Ludwigshafen, Germany. pp. 1-60 (2005)

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