Food Science and Biotechnology

Korean Society of Food Science and Technology (한국식품과학회)
권호 표지

Multiplex PCR Detection of the GT73, MS8xRF3, and T45 Varieties of GM Canola

  • Kim, Jae-Hwan (Institute of Life Sciences and Resources and Graduate School of Biotechnology, Kyung Hee University) ;
  • Kim, Tae-Woon (Institute of Life Sciences and Resources and Graduate School of Biotechnology, Kyung Hee University) ;
  • Lee, Woo-Young (Team of Novel Foods, Korea Food and Drug Administration) ;
  • Park, Sun-Hee (Team of Novel Foods, Korea Food and Drug Administration) ;
  • Kim, Hae-Yeong (Institute of Life Sciences and Resources and Graduate School of Biotechnology, Kyung Hee University)
  • Published : 2007.02.28
Download PDF
⟨ Previous Next ⟩

Abstract

A multiplex polymerase chain reaction (PCR) method was developed to simultaneously detect three varieties of genetically modified (GM) canola. The construct-specific primers were used to distinguish the following three varieties of GM canola; GT73, MS8xRF3, and T45, using multiplex PCR. The FatA (fatty acyl-ACP thioesterase) gene was used as an endogenous canola reference gene in the PCR detection. The primer pair Canendo-FIR containing a 105 bp amplicon was used to amplify the FatA gene and no amplified product was observed in any of the 15 different plants used as templates. The GT73-KHUF1/R1 primer recognized the 3'-flanking region of GT73, resulting in an amplicon of 125 bp. The Barstar-F1/MS8xRF3-R primer recognized the junction region of bars tar and the NOS terminator introduced into MS8xRF3, resulting in a 162 bp amplicon, and the T45-F2/R2 primer recognized the junction region of PAT and the 35S terminator introduced into T45, resulting in an amplicon of 186 bp. This multiplex PCR allowed for the detection of construct-specific targets in a genomic DNA mixture of up to 1% GM canola containing GT73, MS8xRF3, and T45.

Keywords

References

  1. Agbios data base product description (MS8xRF3). Available from: http://www.agbios.com. Accessed May 31, 2006
  2. Smyth S, Phillips PWB. Competitors co-operating: establishing a supply chain to manage genetically modified canola. IFAMR 4: 5166 (2001)
  3. Klee HJ, Muskopf YM, Gasser CS. Cloning of an Arabidopsis thaliana gene encoding 5-enolpyruvylshikimate-3-phosphate synthase: sequence analysis and manipulation to obtain glyphosate-tolerant plants. Mol. Gen. Genet. 210: 437-432 (1987) https://doi.org/10.1007/BF00327194
  4. Bruderer S, Leitner KE. Genetically modified (GM) crops: molecular and regulatory details. Version 2. BATS, 30/06/2003. Available from: http://www.bats.ch/gmo-watch/GVO-report140703.pdf. Accessed May 20, 2006
  5. Miraglia M, Berdal KG, Brera C, Corbisier P, Holst-Jensen A, Kok EJ, Marvin HJP, Schimmel H, Rentsch J, van Rie JPPF, Zagon J. Detection and traceability of genetically modified organisms in the food production chain. Food Chem. Toxicol. 42: 1157-1180 (2004) https://doi.org/10.1016/j.fct.2004.02.018
  6. Duijn G, Biert R, Bleeker-Marcelis H, Boeijen I, Adan AJ, Jhakrie S, Hessing M. Detection of genetically modified organisms in foods by protein- and DNA-based techniques: bridging the methods. J. AOAC Int. 85: 787-791 (2002)
  7. Heo MS, Kim JH, Shin WS, Park SH, Park HK, Kim MC. Limit of detection for genetically modified soybean in doenjang (Korean fermented soypaste). Food Sci. Biotechnol. 13: 657-661 (2004)
  8. Kim YM, Sohn SH, Jeong SI, Yoon MS, Kim TS, Park YH. Detection methods for genetically modified soybeans. J. Korean Soc. Agric. Chem. Biotechnol. 45: 185-189 (2002)
  9. Shin DW, Park SH, Woo GJ, Kim HY, Park CS. Case study for natural gene transfer from genetically modified food to food microorganisms. Food Sci. Biotechnol. 13: 342-346 (2004)
  10. Germini A, Zanetti A, Salati C, Rossi S, Forre C, Schmid S, Fogher C, Marchelli R. Development of a seven-target multiplex PCR for the simultaneous detection of transgenic soybean and maize in feeds and foods. J. Agr. Food Chem. 52: 3275-3280 (2004) https://doi.org/10.1021/jf035052x
  11. Hernandez M, Rodriguez-Lazaro D, Zhang D, Esteve T, Pia, M, Prat S. Interlaboratory transfer of a PCR multiplex method for simultaneous detection of four genetically modified maize events: Bt11, MON810, T25, and GA21. J. Agr. Food Chem. 53: 3333-3337 (2005) https://doi.org/10.1021/jf049192y
  12. Kim JH, Song HS, Heo MS, Lee WY, Lee SH, Park SH, Park HK, Kim MC, Kim HY. Detection of eight different events of genetically modified maize by multiplex PCR method. Food Sci. Biotechnol. 15: 148-151 (2006)
  13. Heo MS, Kim JH, Park SH, Woo GJ, Kim HY. Detection of genetically modified maize by multiplex PCR method. J. Microbiol. Biotechn. 14: 1150-1156 (2004)
  14. Demeke T, Giroux RW, Reitmeier S, Simon SL. Development of a polymerase chain reaction assay for detection of three canola transgenes. J. Am. Oil Chem. Soc. 79: 1015-1019 (2002) https://doi.org/10.1007/s11746-002-0595-2
  15. James D, Schmidt AM, Wall E, Green M, Masri S. Reliable detection and identification of genetically modified maize, soybean, and canola by multiplex PCR analysis. J. Agr. Food Chem. 51: 5829-5834 (2003) https://doi.org/10.1021/jf0341159
  16. Holst-Jensen A, Ronning SB, Lovseth A. PCR technology for screening and quantification of genetically modified organisms (GMOs). Anal. Bioanal. Chem. 375: 985-993 (2003) https://doi.org/10.1007/s00216-003-1767-7