Presentation gmo
- 1. Presence of CaMV35S and NOS in Carica Papaya, Glycine Max, Triticum spp. and ZeamaysValeria Rivera-TorresCarlos Santos-PerezJuan Feliciano-FigueroaRISE StudentsMentor: Dr. Belinda Roman-Aviles
- 2. IntroductionGenetically Modified Organism (GMO) refers to animals, plants and microorganisms that are inserted with a foreign gene or genes, that partially or completely change their metabolic pathway. A GM plant contains a promoter and a terminator in order to create the modification in the DNA sequence of the organism. Promoter-35S Terminator- NOS PCR vs. ELISA
- 3. ObjectivesThis experiment focuses on detecting the presence of genetically modified organisms using food items to extract DNA, amplify using PCR and run a gel electrophoresis.Carica papaya (Papaya)Glycine max (Soy milk)Triticum spp. (Wheat)Zeamays (Whole Corn)
- 4. HypothesisIt is expected that Glycine max and Zeamayswill be Genetically modified whether Carica papaya and Triticum spp. will not be.
- 5. Methodology
- 6. Extracting DNAMortar and Pestle TechniqueGrind with pestle for at least 2 min to form a slurry.Instagene® (BioRad®)Lysis the cells from this samplePrecipitate the DNAPlace the samples on the hot plate for 5 minutes at 37°C.Denature Proteins
- 7. Polymerase Chain Reaction (PCR)Each tube contain the respective master mixMastermix- greenMastermix-red Each eppendorff tube contains: NGMO PMM (green)NGMO GMM (red)GMO + PMMM (green)GMO + GMM (red)Carica Papaya PMM (green)CaricaPapay GMM (red)Glycine max PMM (green)Glycine max GMM (red)Each tube contain the respective master mixMastermix- greenMastermix-red Each eppendorff tube contains: NGMO PMM (green)NGMO GMM (red)GMO + PMMM (green)GMO + GMM (red)Whole corn PMM (green)Whole corn GMM (red)Wheat PMM (green)Wheat GMM (red)
- 8. Electrophoresis Make the agarose gel3% of AgaraoseAdd 10 microliters of Orange G loading dye (LD) to each sample and mix well.Load 20 microliters of the molecular weight ruler and 20 micro liters of each sample into your gel. Ethidium Bromide Reasons for the addition:Orange G Loading DyeMolecular Weight Ruler
- 10. Carlos
- 11. Valeria
- 12. Juan
- 13. Conclusion The hypothesis can not be accepted or rejected. Human errors are involved and affected the results of the investigation. Pipetting, concentrated DNA, lots of primer, evaporation, and contamination are among the possible errors that occurred during this experiment.
- 14. Future Plans The researchers will make a repetition of the same process in order to improve the results.This research can be apply to the agriculture in Puerto Rico in order to identify which of the Puerto Rican products are GMOs. Interdisciplinary contribution to the Puerto Rican community. The effects of GMOs present in the Puerto Rican diet.
- 15. ReferencesCantamutto, M., & Poverene, M. (2007). Genetically modified sunflower release: Opportunities and risks. Fields Crops Research, 101, 133-134.Jiao, Z., Deng, J., Li, G., & Cai, Z. (2010). Study on the compositional differences between transgenic and non-transgenic papaya(Carica papaya L.). Journal of Food Composition and Analysis, 23, 640-647.Scipioni, A., Saccarola, G., Arena, F., & Alberto, S. (2008). Strategies to assure the abscence of GMO in food products application process in a confectionery firm. Food Control, 16, 569-578.Terzi, V., Malnati, M., Barbanera, M., Stanca, A., & Faccioli, P. (2003). Development of analytical systems based on real-time PCR for Trititcum species- specific detection and quatitation of bread wheat contamination in semolina and pasta. Journal of Cereal Science, 38, 87-94.BioRad® Manual of: Biotechnology Explorer™ GMO Investigator™ Kit
Editor's Notes
- #3: 35S promoter of the cauliflower mosaic virus (CaMV 35S)NopalineSynthase(NOS) gene of Agrobacteriumtumefaciens.Duplex vs. SimplexThere was two different types of procedures in order to identify if a fruit is genetically modified- PCR that is for gene or ELISA- that is for proteins. Actually, a promoter is involved in RNA transcription from a DNA sequence. It regulates the binding of RNA polymerase and the rate at which RNA is transcribed.Promoters are specific sequences of DNA where polymerase initiates transcription. The terminator is a sequence that signals the end of transcription.For translation a AUG is the start codon, and translation stops at the STOP codon.A transgene is any foreign DNA that has been inserted into the genome of an organism, typically using transposable elements.
- #7: Explain 2 grams of each sample.
- #8: Red- primersforterminator(NOS) NopalineSynthaseofAgrobacteriumtumefasiensandpromoter (CamV35-Cauliflowermosaic virus)Greenisforthephotsystem gene identification-Indentifyifthesampoleis a productof a plant.
- #13: Red iswho determineiftheproductis a GMO.