Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Qualitative and Quantitative Analysis of Genetically Modified Pepper

  • Song, Hee-Sung (Institute of Life Science & Resources and Graduate School of Biotechnology, Kyung Hee University) ;
  • Kim, Jae-Hwan (Institute of Life Science & Resources and Graduate School of Biotechnology, Kyung Hee University) ;
  • Kim, Dong-Hern (Division of Biosafety, National Institute of Agricultural Biotechnology) ;
  • Kim, Hae-Yeong (Institute of Life Science & Resources and Graduate School of Biotechnology, Kyung Hee University)
  • Published : 2007.02.28
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Abstract

For the development of qualitative and quantitative PCR methods of genetically modified (GM) pepper developed in Korea, a capsanthin-capsorubin synthase (CCS) gene was used as the endogenous reference gene. The primer pair ccs-F/R amplifying the pepper endogenous gene gave rise to an amplicon of 102 bp. No amplified product was observed when DNA samples from 16 different plants were used as templates. The construct-specific primer pairs amplifying the junction region of the bar gene and Ti7 introduced in GM pepper gave rise to an amplicon of 182 bp. Quantitative PCR assay was performed using a TaqMan probe and a standard plasmid as a reference molecule, which contained both an endogenous and event-specific sequence. For the validation of this method, the test samples containing 0.1, 1, 3, 5, and 10% GM pepper were quantified.

Keywords

References

  1. Guideline of Detection Methods of Genetically Modified Foods; Korea Food and Drug Administration: Seoul, Korea, 2000
  2. Haung, H. Y. and T. M. Pan. 2004. Detection of genetically modified maize MON810 and NK603 by multiplex and realtime polymerase chain reaction methods. J. Agric. Food Chem. 52: 3264-3268 https://doi.org/10.1021/jf049944o
  3. Hernández, m., A. Rio, T. Esteve, S. Prat, and M. Pla. 2001. A rapeseed specific gene, acetyl-CoA carboxylase, can be used as a reference for qualitative and real-time quantitative PCR detection of transgenes from mixed food samples. J. Agric. Food Chem. 49: 3622-3627 https://doi.org/10.1021/jf010173n
  4. Huang, C. C. and T. M. Pan. 2005. Event-specific realtime detection and quantification of genetically modified roundup ready soybean. J. Agric. Food Chem. 53: 3833- 3839 https://doi.org/10.1021/jf048580x
  5. Huh, J. H., B. C. Kang, S. H. Nahm, S. Kim, K. S. Ha, M. H. Lee, and B. D. Kim. 2001. A candidate gene approach identified phytoene synthase as the locus for mature fruit color in red pepper (Capsicum spp.). Theor. Appl. Genet. 102: 524-530 https://doi.org/10.1007/s001220051677
  6. Hupfer C., H. Hotzel, K. Sachse, F. Moreano, and K. H. Engel. 2000. PCR-based quantification of genetically modified Bt maize; single-competitive versus dual-competitive approach. Eur. Food Res. Technol. 212: 95-99 https://doi.org/10.1007/s002170000208
  7. James, C. 2003. Preview: Global status of commercialized transgenic crops. ISAAA Briefs, 30
  8. Jung, S. J., H. J. Kim, and H. Y. Kim. 2005. Quantitative detection of Salmonella typhimurium contamination in milk, using real-time PCR. J. Microbiol. Biotechnol. 15: 1353- 1358
  9. Kim, J. H., H. S. Song, D. H. Kim, and H. Y. Kim. 2006. Quantification of genetically modified canola GT73 using TaqMan real-time PCR. J. Microbiol. Biotechnol. 16: 1778- 1783
  10. Kim, Y. H., S. J. Choi, H. A. Lee, and T. W. Moon. 2006. Quantitation of CP4 5-enolpyruvylshikimate-3-phosphate synthase in soybean by two-dimensional gel electrophoresis. J. Microbiol. Biotechnol.16: 25-31
  11. Kuribara, H., Y. Shindo, T. Matsuoka, K. Takubo, S. Futo, N. Aoki, T. Hirao, H. Akiyama, and Y. Goda. 2002. Novel reference molecules for quantitation of genetically modified maize and soybean. J. AOAC Int. 85: 1077-1089
  12. Lee, S. H., H. J. Park, S. M. Cho, H. K. Chun, D. H. Kim, and T. H. Ryu. 2004. Comparison of major nutrients and mineral contents in genetically modified herbicide-tolerant red pepper and its parental cultivars. Food Sci. Biotechnol. 13: 830-833
  13. Lee, S. H., S. H. Kang, Y. H. Park, D. M. Min, and Y. M. Kim. 2006. Quantitative analysis of two genetically modified maize lines by real-time PCR. J. Microbiol. Biotechnol. 16: 205-211
  14. Matsuoka, T. 2001. GMO labeling and detection methods in Japan. APEC-JIRCAS Joint Symposium and Workshop on Agricultural Biotechnology
  15. Notification No. 1775; Food and Marketing Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan: Tokyo, Japan, 2000
  16. Notification No. 2000-31; Ministry of Agriculture and Forestry of Korea: Seoul, Korea, 2000
  17. Permingeat, H. R., M. I. Reggiardo, and R. H. Vallejos. 2002. Detection and quantification of transgenes in grains by multiplex and real-time PCR. J. Agric. Food Chem. 50: 4431-4436 https://doi.org/10.1021/jf020081d
  18. Regulation (EC) No. 1829/2003 of the European Parliament and of the Council
  19. Testing Manual for Genetically Modified Agricultural Products; National Agricultural Quality Management Services of Korea: Seoul, Korea, 2002
  20. Weng, H., L. Yang, Z. Liu, J. Ding, A. Pan, and D. Zhang. 2005. Novel reference gene, high-mobility-group protein I/ Y, used in qualitative and real-time quantitative polymerase chain reaction detection of transgenic rapeseed cultivars. J. AOAC Int. 88: 577-584
  21. Yang, L., S. Xu, A. Pan, C. Yin, K. Zhang, Z. Wang, Z. Zhou, and D. Zhang. 2005. 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: 9312-9318 https://doi.org/10.1021/jf051782o