Genetically Modified Organisms (Transgenics) – “The Right to Know” Princeton Satellite 2013 Kathy McKone What does it mean to be genetically modified? Inserting a desired gene from one organism into an organism of a different species. WHY Bother w/ GM Food? 1980 World Population = 4.5 Billion 2011 World Population = 7 Billion Farmable Land disappearing Worries about ability to produce enough food Concerns of pesticide & herbicide overuse; and long-term effects of these chemicals The Workhorse of Genetic Engineering = Bacteria General GMO: http://www.youtube.com/watch?v= jAP6ZtfP9ZQ More Advanced: http://www.youtube.com/watch?v=T-IJikX1144 BioRad Intro: http://www.youtube.com/watch?v=RIP sjrXMMmw http://www.youtube.com/watch?v=m39 25Pw-VwU Genetically Engineering Insulin Which foods contain GM product? US Approval for GM food crops •Corn •Soy •Papaya •Canola •Potato •Chicory •Rice •Squash •Sugarbeet •Tomatoes Approval does not necessarily mean these crops are distributed Database of GM crops: www.agbios.com Agriculture: Genetically Modified (GM) Organisms Inserting genes from one organism (Bacteria) into another organism (Plant). Adding genes to plants can make them draught or insect resistant. 1994: first GM food was sold: Flavr Savr tomato = able to ripen on the vine & not turn to mush; wasn’t successful b/c of taste Image:Results of insect infestation on Bt (right) and non-Bt (left) cotton bolls. Source: USDA According to the USDA, in the U.S. in 2011: CROPS genetically modified: 94% soy 88% corn 90% cotton Which item would NOT be a good selection to test with students? Which foods contain GM product? % of all crop planted 100 90 GM corn GM soy 80 70 60 50 40 30 20 10 19 95 19 96 19 97 19 98 19 99 20 00 20 01 20 02 20 03 20 04 0 Sources: 1996-1999 Fernandez and McBride, 2000-2004: USDA, National Agriculture Statistics Service, Acreage. GM Foods are NOT required to be labeled in US. California's Prop 37 – to label GM foods *DEFEATED (Nov. 2012) Biotech companies like Monsanto and Syngenta hold patents on the GE seeds and also "control who conducts research". http://www.aljazeera.com/indepth/opinion/20 12/11/201211811503032830.html *Monsanto, Pesticide Companies Spend $7.1 Million to Defeat Right to Know GMO Labeling Measure in CA http://www.carighttoknow.org/5_million_more Which foods yield viable plant DNA? Very Reliable Reliable Fresh corn Veggie sausages Veggie burgers Fresh papaya Tortilla chips Fried corn snacks Salad dressing Corn bread mix Flavored tortilla chips Popcorn Cereal (eg cornflakes) Corn meal Puffed corn snacks Fries Wheat flour Soy flour Meatballs and burgers containing soy protein Potato chips Soy-based protein drinks/powders Less Reliable Very Difficult / Not Possible Oil Cell Biology REVIEW: Three Segments of a Gene 1) Promoter = found at the beginning of a gene; docking site of RNA polymerase 2) Gene = codes for protein of interest (pest resistance) 3)Terminator = stop transcription Why use CaMV 35S and NOS? CaMV 35S – Sequence for the promoter of 35S transcript of the Cauliflower mosaic virus. Used because it functions in every plant cell NOS- Sequence for nopaline synthase terminator from soil bacterium Agrobacterium tumefacians Used because it evolved to be recognized in most plants 65% of food crops use CaMV promoter; adding NOS detection ~ 85% Most Used Promoter used in GM crops The Cauliflower Mosaic Virus promoter (CaMV 35S) is used in most transgenic crops to activate foreign genes which have been artificially inserted into the host plant http://www.microbiologyonline.org.uk/graphics/bluetong.gif Popular gene from soil bacterium: Bacillus thuringiensis (Bt) Bt gene protein endotoxin (lethal to many insects) Pesticides are not needed for crops! Endotoxins are activated by the HIGHLY ALKALINE insect midgut. Also kills non-target insects (monarch butterfly). Desirable Traits Pest Resistance Herbicide Tolerance Viral Resistance Drought Resistance Increased Nutritional Value; golden rice (beta carotene) Improved Fruit Altered Ripening Opponents argue Creation of super pests Creation of super weeds Loss of biodiversity Biotechnology companies control agriculture GMO-Based Web Sites Cornell http://www.geopie.cornell.edu University of Nebraska http://croptechnology.unl. edu/ Pro-GMO http://www.monsanto.c om Anti-GMO http://www.greenpeace.org BioRad’s GMO Investigator Kit Contents Not Included but required: •Thermal cycler •Water bath/heat block •Electrophoresis Module (agarose, TAE buffer & Fast Blast DNA stain) •Electrophoresis equipment & power supply •2-20 ul pipettes & barrier tips Bio-Rad certified Non-GMO food InstaGene Master Mix GMO primers Plant PSII primers GMO & PSII positive control DNA PCR MW Ruler DPTPs, microtubes, PCR tubes, foam floats Manual