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GMOs in the economy 30. The GMO debate „….don’t take them to your mouth!” Keyword: natural Chance and instincts Genetically Modified Foods 31. Flavr-Savr: the first license for genetically modified food NORMAL PROCESS PG protein DNA MODIFIED PROCESS PG mRNA DNA asRNA PG gene: play a role in ripening - inhibition: increased storage Genetically Modified Crops 32. 1st and 2nd Generation GMOs Rice (Golden rice) - high β-carotene level - 2 genes from lent-lily - 1 gene from a bacterium rape - high oil content - 1 gene from a bacterium Maize, - insect resistance - Herbicide resistance - Bacillus thüringiensis gene soybean Cotton - insect resistance - Pesticide resistance Sugar beet - Pesticide resistance 2nd generation 1st generation Genetically Modified Foods 33. 3rd Generation GMOs: plant factories Potato Spinage - Vaccine against hepatitis B - Vaccine against rabies Tobacco - Against caries - insulin maize - lactoferrin Arabidopsis - Humán intrinsic factor (against anemia) Arabidopsis Terminator Traitor technologies 33b. Terminator: GMOs produce sterile seeds Traitor: the development of GMO has to be activated Advantages: (1) Protection of proprietary right (purchusing new seeds yearly) (2) Protection of spread in nature 34. How to modify plants? Cold-tolerant tomato Agrobacterium Ti plasmid 1. Plaice antifreeze protein 2. Cloning of anti-freeze protein gene to Ti-plasmid of Agrobacterium 3. Recombinant Agrobacterium infects tomato tissues Promoter Promoter 4. Antifreeze-protein expressing transgenic tomato Ti plasmid T-DNA 5. Antibiotics resistance A 3rd generation GM animal 34. Human ATIII gene cloning Anti-coagulant protein in goat milk X Gene injection to zygote 2. Fertilized egg cell Integration of ATIII gene to the genome ATIII protein in goat milk 3. 4. Transgenic embryo develops in the surrogate mother Genetically Modified Livestock Transgenic salmon grows faster Higher level of beta and kappa caseins in the milk of transgenic cow Various human transgenes in pigs for transfer of liver to human 36. 37. Genetically Modified Human Two major aims: (1) Gene therapy (2) Cosmetic improvement Genetically modified microorganisms Viruses gene therapy, tumor therapy, circuit tracing, vaccines Bacteria recombinant proteins Fungi: fermentation, recombinant proteins 38. Recombinant proteins Recombinant proteins Disease Insulin: Clotting factors: Growth hormones: diabetes hemophilia dwarfism Advantage: 39. It does not cause disease (formerly they were gained from corps) Disadvantage: The processing of proteins are different in bacteria (e.g. there is no glycosylation) 40. Bacterial DNA Non-integrating plasmids Genomial DNA Integrating plasmid Genomial DNA Cloning (if foreign DNA is inserted) 41. The effects of insulin glycogen synthseis fatty acid synthesis glycolysis 42. Recombinant insulin Human cell Bacterium cell plasmid DNA RE digestion RE digestion Human insulin gene Protein engineering (1) Directed Evolution (2) Rational Protein Engineering 43. Antibody techniques 44. Polyclonal antibodies Antigen injected into the animal body B cell Memory B cells Antigen B cell Plasma cells Polyclonal antibodies Polyclonal antibodies containing serum is taken out from the animal’s blood 45. 46. Monoclonal antibodies Isolation of B cells from the spleen Cultivation of myeloma cells Immunization of a mouse Fusion of myeloma and B cells Separation of cell lines Screening of suitable cell lines Multiplication in vitro or in vivo Harvesting Inject mouse with antigen X Monoclonal antibodies Mouse spleen cells; some cells ( ) make antibody to antigen X Mutant mouse myeloma cells unable to grow in HAT medium Mix and fuse cells Transfer to HAT medium Unfused cells ( Fused cells ( ) die ) grow Culture single cells in a separate wells Test each well for antibody to antigen X 47. Production of hybrid cells Selective medium, only hybrid cells proliferate Cell fusion Plasma cell Tumor cell (hybridoma) Heterocaryon Hybrid cell 48. Recombinant antibodies Enzyme papain Antibody gene bacteriophage gIII gene Fab fragment gIII proteins (antigen binding) Fc fragment Disulfide bond (responsible for physiological effects) Antibody Fab fragment 49. 50. Antibiotics bacterium mold Alexander Fleming The penicillin inhibits the growth of bacteria 51. Antibiotics The effects of antibiotics How bacteria defend themselves? antibiotic Structure and function of DNA (Nitrimidazole)