Qualitative PCR Detection of GM rices (Milyang 204 and Iksan 483) developed in Korea

국내에서 개발된 GM 쌀 (밀양 204호, 익산 483호)에 대한 정성 PCR 분석법 개발

  • Kim, Jae-Hwan (Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Song, Hee-Sung (Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Jee, Sang-Mi (Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Ryu, Tae-Hun (Division of Biosafety, National Institute of Agricultural Biotechnology) ;
  • Kim, Dong-Hern (Division of Biosafety, National Institute of Agricultural Biotechnology) ;
  • Kim, Hae-Yeong (Department of Food Science and Biotechnology, Kyung Hee University)
  • 김재환 (경희대학교 생명공학원) ;
  • 송희성 (경희대학교 생명공학원) ;
  • 지상미 (경희대학교 생명공학원) ;
  • 류태훈 (농촌진흥청 농업생명공학연구원 생물안전성과) ;
  • 김동헌 (농촌진흥청 농업생명공학연구원 생물안전성과) ;
  • 김해영 (경희대학교 생명공학원)
  • Published : 2005.12.31

Abstract

For the development of qualitative PCR detection method of genetically modified rice (Oryza sativa L.), rice species specific gene, SAMDC1 (S-adenosylmethionine decarboxylase), was selected and validated as suitable for use as an endogenous reference gene in rice. The primer pair OsSAMDC1-5'/3' with 110 bp amplicon was used for amplification of the rice endogenous gene, SAMDC1 and no amplified product was observed from 19 different plants as templates. Qualitative PCR method was assayed with 2 different GM rices (Milyang 204 and Iksan 483) developed in Korea. For the qualitative PCRs, the construct-specific detection primer pairs were constructed. Os204-5'/OsNOS-3' amplifying the junction region of GUS gene and NOS terminator introduced in Milyang 204 gave rise to an amplicon 172 bp; also, Os483-5'/OsNOS-3' amplifying the junction region of Bar gene and NOS terminator introduced in Iksan 483 gave rise to an amplicon 161 bp.

국내에서 개발된 GM 쌀인 밀양 204호, 익산 483호의 정성 PCR 분석법의 개발을 위해 쌀의 내재 유전자로써 SAMDC1(S-adenosylmethionine decarboxylase)에 특이적인 primer (OsSAMDC1-5'/3')쌍을 제작하여 쌀을 포함한 20개 작물에 대해 PCR을 수행하여 쌀에 특이적으로 증폭되는 것을 확인하였다. 또한, 밀양 204호에 삽입된 GUS 유전자와 NOS terminator 연결 부위를 증폭시켜 172 bp의 PCR 산물을 얻을 수 있는 primer(Os204-5'/OsNOS-3')와 익산 483호에 삽입된 bar 유전자와 NOS terminator 연결 부위를 증폭시켜 161 bp의 PCR 산물을 얻을 수 있는 primer(Os483-5'/OsNOS-3')을 이용하여 국내 개발된 GM 쌀인 밀양 204호, 익산 483호의 PCR 정성 분석법을 확립하였다.

Keywords

References

  1. Clive, J. (2004) Preview: Global status of commercialized biotech/GM crops. ISAAA briefs No. 32, ISAAA: Ithaca, NY
  2. Huang, J. K., Hu, R, Rozelle, S. and Pray, C. (2005) Insect-resistant GM rice in farmers' fields: Assessing productivity and health effects in China. Science 308, 688-690 https://doi.org/10.1126/science.1108972
  3. High, S. M., Cohen, M. B., Shu, Q. Y. and Altosaar, I. (2004) Achieving successful deployment of Bt rice. Trends in Plant Sci. 9, 286-292 https://doi.org/10.1016/j.tplants.2004.04.002
  4. Oard, J. H., Linscombe, S. D., Braverman, M. P., Jodari, F., Blouin, D. C., Leech, M., Kohli, A., Vain, P., Cooley, J. C. and Christou, P. (1996) Development, field evaluation and agronomic performance of transgenic herbicide resistant rice. Mol. Breed 2, 359-368 https://doi.org/10.1007/BF00437914
  5. Wu, G, Cui, H., Ye, G, Xia, Y., Sardana, R, Cheng, X, Li, Y., Altosaar, I. and Shu, Q. (2002) Inheritance and expression of the crylAb gene in Bt (Bacillus thuringiensis) transgenic rice. Theor. Appl. Genet. 104, 727-734 https://doi.org/10.1007/s001220100689
  6. Won, Y. J., Yi, G. H., Cho, J. H., Ko, J. M., Park, H. M., Han, C. D., Yang, S. J., Kim, S. C. and Nam, M. H. (2004) Establishment of a new breeding scheme for rapid release of variety using bar gene transformed rice. Korean J. Plant Biotechnol. 31, 7-11
  7. Zi, X. (2005) GM rice forges ahead in China amid concerns over illegal planting. Nat. Biotechnol. 23, 637 https://doi.org/10.1038/nbt0605-637
  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. 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
  10. 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.
  11. Li, J. Y. and Chen, S. Y. (2000) Differential accumulation of the S-adenosylmethionine decarboxylase transcript in rice seedlings in response to salt and drought stresses. Theor. Appl. Genet. 100, 782-788 https://doi.org/10.1007/s001220051352
  12. McElroy, D., Blowers, A. D., Jenes, B. and Wu, R. (1991) Construction of expression vectors based on the rice actin 1. 5' region for use in monocot transformation. Mol. Gen. Genet. 231, 150-160 https://doi.org/10.1007/BF00293832
  13. Uzogara, S. G. (2000) The impact of genetic modification of human foods in the 21st century. Biotechnol. Adv. 18, 179-206 https://doi.org/10.1016/S0734-9750(00)00033-1