Detecting Transgenes in Soy: A Study of GMOs in Soy Products

1. Who’s Bean Modifying Our Soy: The Detection of Transgenes in Soy Products Courtney Geary, Christian Schill, Yuge Xiao, and Sydney Yang http://www.healthbeyondhype.com/info/wp-content/uploads/2011/05/Soy-field-by-eduardoMoraes.jpg

2. http://www.apoorvasaxena.in/Gallery.html Project Summary • What constitutes a genetic modification (GM)? • Labeling • Techniques • Genetic Assays • Protein Based Assays • Data Analysis • Conclusion

3. GM Foods Across the Globe • United States: biggest manufacturer for GM foods Labelling • United States does not require labelling8 • Non-GMO Project and USDA Certified Organic6 http://gmoanswers.com/public-review

4. http://sitemaker.umich.edu/sec005group5/files/gmo_3.gif GM crops and attitudes towards GM foods across the world Labelling

5. How are GM Crops Created? • Gene isolated and inserted into gene cassette (promoter, target gene, terminator)5 • Gene cassette inserted into plant through gene gun or agrobacterium5 http://www.scq.ubc.ca/wp-content/uploads/2006/07/genetransferring.gif

6. Genetic Modification of Soy Plants: http://www.24hourturnaround.com/soy-debate/ • Soy plants are the most commonly genetically modified plant in the United States. COMMON SOY INSERTIONS OF INTEREST: • CaMV35S Promoter2 • NOS Terminator • NPTII9 • CP4-EPSPS “Roundup Ready”3 SOY LECTIN (control): • All soy plants produce soy lectin.1

7. Our Study... Locally Grown Soybeans (ARA) http://alturl.com/8e8ei Testing for presence of GMOs in these soy based products: Pepperidge Farms Goldfish Crackers (GC) http://alturl.com/vhp6s House Foods Premium Tofu (HFT) Savoritz 4 Kids Cheddar Penguins (PCC) http://alturl.com/sqr7a http://alturl.com/gki2x

8. How Are GMOs Detected? There are many methods that can be used to detect GMOs. Gel Electrophoresis ELISA Testing Polymerase Chain Reaction (PCR) http://www.antidopingresearch.org/PressRoom_images_illustrations.php

9. How Does PCR Work? Simplex & Multiplex PCR • Detect the presence of and amplify target gene(s)4 • Uses unique primer(s) • Includes junction primers Primer Primer http://www.scielo.br/img/revistas/bjm/v34n4/4a01f02.gif

10. Gel Electrophoresis • Cathode attracts negatively charged DNA4 • Smaller fragments travel longer distance from the wells4 • Sizes of bands can be calculated http://www.britannica.com/EBchecked/media/40224/In-gel-electrophoresis-an-electric-field-is-applied-to-a

11. ELISA Tests for Proteins • Uses antibodies to detect proteins in a solution • Antibodies bind only to specific proteins • Bound protein reacts to yield colored product http://exploreable.wordpress.com/2011/05/25/elisa-enzyme-linked-immunosorbent-assay/

12. Methods • DNA extracted from all samples • UV-Vis Spectroscopy to determine purity • Simplex PCR to detect CaMV • Multiplex PCR to detect CaMV, Nos, CP4-EPSPS, and NPTII • Gel Electrophoresis on all PCR products • ELISAs to detect the presence of proteins (CP4-EPSPS, NPTII) http://www.apoorvasaxena.in/Gallery.html

13. Data Analysis http://www.worldwildlife.org/projects/advancing-responsible-lending

14. Results of UV/Vis Spectroscopy: Ideal Acceptable Poor

15. Simplex PCR - Gel Electrophoresis bp bp bp bp

18. NPTII ELISA • inconclusive • mixed positive and negative absorbances • problem might lie in shared factor Multiplex PCR • inconclusive • no visible bands • possible causes: contamination, impurities6 • not primary sources • too many primers

19. Pepperidge Farms Goldfish Crackers (GC) • no CamV 35S band • low RR protein (0%-0.1%) • Tertiary Source AsaROW AG390 (ARA) • presence of CamV 35S band • > 2% RR protein content • Primary Source Penguin Cheddar Crackers (PCC) • no CamV 35S band • no RR protein present • Tertiary Source • made in Canada • product of Aldi, of Germany House Foods Premium Tofu (HFT) • no CamV 35S band • low RR protein concentration (0%-0.1%) • Secondary Source • consistent with Non-GMO Project Verified label

20. Conclusion • Genetically modified foods have become increasingly more prevalent in the American diet because of the lack of labeling requirements. • This study focused on the detection of GM soy in four products; Cheddar Penguins, Goldfish Crackers, Tofu, and Soy Beans. • Of the four products, only the soybeans were found to contain genetic modification common in soy.

21. Sources (1) Andrews, Ryan. "All about Lectins." www.precisionnutrition.com. N.p., n.d. Web. 2014. <http://www.precisionnutrition.com/all-about-lectins>. (2) “The CaMV 35S Promoter" Patentlens.net. Cambia, n.d. Web. 27 July 2014. (3) Funke, Tod, Huijong Han, Martha L. Healy-Fried, Markus Fischer, and Ernst Schönbrunn. "Molecular Basis for the Herbicide Resistance of Roundup Ready Crops." Molecular Basis for the Herbicide Resistance of Roundup Ready Crops. N.p., n.d. Web. 27 July 2014. (4) James, Delano, et al. "Reliable Detection and Identification of Genetically (5) Genetically Modified Crops: Resources for Environmental Literacy. Arlington: NSTA, 2007. PDF file. Modified Maize, Soybean, and Canola by Multiplex PCR Analysis." Journal of Agriculture and Food Chemistry (2003): n. pag. Print. (6) <http://www.nongmoproject.org/learn-more/understanding-our-seal/> (7) Oswald, Nick. "The Essential PCR Troubleshooting Checklist." Bitesize Bio. N.p., 28 Jan. 2008. Web. 30 July 2014. <http://bitesizebio.com/343/ the-essential-pcr-troubleshooting-checklist/>. (8) Rabin, Roni Caryn. "Information Not on the Label." The New York Times 26 May 2014: n. pag. New York Times. Web. 30 July 2014. <http://well.blogs.nytimes.com/2014/05/26/information-not-on-the-label/ ?_php=true&_type=blogs&_r=0>. (9) Roa-Rodriguez, Carolina, and Carol Nottenburg. "Antibiotic Resistance Genes and Their Uses in Plant Genetic Transformation." Patentlens.net. Cambia, 2003. Web. 17 July 2014.

22. ACKNOWLEDGEMENTS • Pennsylvania Department of Education • Corporate Sponsors • Individual Donors • PGSS Alumni Association • Nathan Wilson, Team Project Leader • Charanya Kaushik, Team Project TA • Dr. Carrie Doonan • Barry Harris • Dr. Barry Luokkala • Core and Elective Course Professors • RLDs and TAs