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: jAP6ZtfP9ZQ jAP6ZtfP9ZQ More Advanced: BioRad Intro: sjrXMMmw 25Pw-VwU 25Pw-VwU Genetically Engineering Insulin

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: Which foods contain GM product?

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? Sources: Fernandez and McBride, : USDA, National Agriculture Statistics Service, Acreage.

GM Foods are NOT required to be labeled in US. 12/11/ html 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". *Monsanto, Pesticide Companies Spend $7.1 Million to Defeat Right to Know GMO Labeling Measure in CA

Very ReliableReliableLess ReliableVery Difficult / Not Possible Fresh cornVeggie sausagesVeggie burgersOil Fresh papayaTortilla chipsFried corn snacksSalad dressing Corn bread mixFlavored tortilla chips PopcornCereal (eg cornflakes) Corn mealPuffed corn snacks FriesWheat flour Soy flourMeatballs and burgers containing soy protein Potato chips Soy-based protein drinks/powders Which foods yield viable plant DNA?

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

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 mid- gut. 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 pie.cornell.edu University of Nebraska edu/ Pro-GMO om om Anti-GMO

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