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Biotechnology CLIL module
- 1. Biotechnology CLIL MODULE ABOUT… Author:Prof.ssa Elena Bruno
- 2. UNIT 1 CONTENTS • What’sbiotechnology? • How old is biotechnology? • Differences between traditional and innovative biotechnology • Recombinant DNA technology • Production of insulin with recombinant DNA technology
- 3. What is biotechnology? Biotechnologyis the use of living cells or organisms to produce materials useful to people. Traditional Biotechnology Innovative Biotechnology Use of living beings and selection of natural features in living beings use of genetically engineered microorganisms, plant and animal cells GMO
- 4. How Old IsBiotechnology? 10,000 BC Domesticating Crops 1880’s Production of Vaccines Domesticating Animals 8,000-9,000 BC 4,000 BC Leavening Bread 6,000 BC Brewing Beer 1940’s Production of Antibiotics 1980’s Use of genetically modified organisms molds yeasts
- 5. Do traditional biotechnology modifyliving beings? Old biotechnology modifies living beings by selective breeding of plants/animals: on the contrary of modern biotechnology, scientists do not directly modify the DNA
- 6. Do traditional biotechnology modifyliving beings? The traditional biotechnology also use radiations (such as X-rays) to increase the rate of mutations in the DNA of a species (such as wheat): in this way it is possible to increase the genetic variability in a short time and to obtain favorable variations. For example the variety "wheat Creso" was obtained in this way. irradiated wheat is not a GMO!
- 7. Do innovative biotechnology modifiesliving beings? Modern biotechnology modify genetic material (DNA) in a way that does not occur naturally by mating and/or natural recombination EXAMPLE: Resistance against insects is achieved by incorporating into the food plant the gene for toxin production from the bacterium Bacillus thuringiensis (Bt) CORN COB
- 8. Applications of biotechnology MedicalBiotechnology • Diagnostics • Therapeutics • Vaccines Agricultural Biotechnology • Plant agriculture • farms • Food processing Environmental Biotechnology • Cleaning through bioremediation • Preventing environmental problems Forensic Biotechnology • Paternity test • Scientific investigations Industrial Biotechnology • Energy production • New materials
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- 10. Some tools ofbiotechnology Recombinant DNA technology Gene Therapy Cloning Monoclonal Antibodies’ production Stem cells PCR DNA fingerprint DNA sequencing In vitro fertilization ……AND MORE!!!!
- 11. Recombinant DNA technology RecombinantDNA technology is one of the recent advances in biotechnology, which was developed by two scientists named Boyer and Cohen in 1973. Recombinant DNA technology works by taking DNA from two different sources and combining that DNA into a single molecule. That alone, however, will not do much. Recombinant DNA technology only becomes useful when that artificially-created DNA is reproduced. This is known as DNA cloning.
- 12. What’s recombinant DNA? RECOMBINANTDNA : is a DNA molecule that has been made in the laboratory using at least two different sources of DNA Recombinant DNA is produced using in vitro molecular techniques to isolate and manipulate fragments of DNA ? To produce Recombinat DNA you need Molecular scissors: Restriction enzyme Molecular glue: Ligasi enzyme A vector: Plasmide, viruses, etc
- 13. Restriction enzymes Enzymes thatcleave DNA are used to manipulate and recombine DNA. Restriction enzymes, naturally used by bacteria as defense against bacteriophages, cut DNA into smaller pieces. Restriction enzymes cut DNA at specific base sequences called restriction site. CUT=CLEAVE
- 14. Restriction enzymes Some restriction enzymes recogniseand cut palindromic DNA sequences (sequences that read the same way in both directions). Some of them make staggered cuts, generating fragments with sticky ends
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- 16. 16 Enzyme ligase The stickyends may join: a fragment from one source, such as human, can be joined to a fragment from another source, such as bacterium. Initially the fragments are held together only by weak hydrogen bonds: for these reason you use the enzyme ligase to catalyze the formation of covalent bonds between adjacent nucleotides at the ends of the fragments, joining them to form a single, larger molecule They join ligase
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- 18. 18 Plasmids A plasmid isa small DNA molecule within a cell that is physically separated from a chromosomal DNA and can replicate independently. They are most commonly found in bacteria as small, circular, DNA molecules • Plasmids are naturally exchanged among organisms • They can be modified and used to deliver new pieces of DNA into an organism=VECTOR They often contain genes for antibiotic resistance http://education-portal.com/academy/lesson/what-is-a-dna-plasmid-importance-to-genetic-engineering.html#lesson
- 19. Six steps ofrecombinant DNA technology 1. Isolating (vector and target gene) 2. Cutting (Cleavage) 3. Joining (Ligation) 6. Selecting (Screening) 5. Cloning 2 3 1 4 4. Transformation (Introduction of the plasmid into cells) 5 6
- 20. Six steps ofrecombinant DNA technology
- 21. Screening ampicilline resistance gene lacZgene unfunctioning lacZ gene human gene Plate out the transformed bacteria on a solid nutrient medium containing ampicillin and a sugar called X-gal. Only transformed bacteria that have the ampicillin- resistance plasmid will grow. This bacterium express lacZgene, will hidrolyse sugar and form BLUE colony This bacterium DOESN’T express lacZgene, it isn’t able to hidrolyse sugar, will form WHITE colony: it’s a GMO with useful external gene Repoter genes or marker genes
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- 23. Production of insulinin the past Type I diabetes mellitus is a metabolic disease caused by lack of hormone insulin. It’s treatable only with injection In the past, insulin was taken from the pancreases of cows and pigs. The method was expensive and some patients had severe allergic reaction to the foreign insulin or its contaminants these problems have been overcome thanks to a GMO http://education-portal.com/academy/lesson/what-is-genetic-engineering-definition-and-examples.html
- 24. 24 Production of insulinwith recombinat DNA technology simplified version
- 25. Production of insulinwith recombinat DNA technology Correct version Two genes are chenically synthesized, one for the insulin A chain and one for the Insulin B chain. Plasmid are extracted from Escherichia coli. To make the bacteria produce insulin, the insulin gene must be linked to the beta-galactosidase gene, which carries a promoter for tanscription. The recombinant plasmids are inserted back into bacteria, these bacteria reproduce so that Insulin gene are cloned. The GMO bacteria produce the two Insulin chains that are extracted, purufied and mixed together . The A and B insulin chains conncet via disulfade cross linkages to form the functional insulin protein
- 26. 26 Animals are geneticallyengineered to exalt specific traits or obtain useful products: •GM animals can produce pharmaceuticals •GM mice are bred for human disease research •Xenotransplantation (animals as organs donor in transplantation) Genetically modified animals
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- 29. UNIT 2 CONTENTS • Biotecchnologyin medicine: production of drugs • Monoclonal antibodies • Stem cells (blended learning) • Gene therapy
- 30. TPAis a naturallyoccuring human protein that dissolves blood clots. It’used to treat patients suffering from blood clotting in heart attack or strokes Production of drugs
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- 32. Production of drugswith pharming An expression vector carrying a desired gene can be put into an animal fertilized egg (embryo), which is implanted into a surrogate mother. The transgenic offspring produce the new protein in their milk. The milk is easily harvested and the protein isolated, purified, and made clinically available to patients. Another way of making medically useful products in large amount is pharming: the production of pharmaceuticals in farm animals or plants 2. The human transgene is injected into the Fertilized eggs 3. Eggs are transferred to recipient ewes 1. Donor ewes: after insemination, fertilized eggs are collected 4. The offspring are raised- selection of animals producing the human protein in the milk Ewe=female sheep
- 33. Production of drugswith pharming • Human growth hormone (for children suffering deficiencies) can now be produced by transgenic cows. • Only 15 such cows are needed to supply all the children in the world suffering from this type of dwarfism. transgenic animals are not well seen by the public, and yet ....
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- 35. Use of Monoclonalantibodies
- 36. Use of Monoclonalantibodies
- 37. ADC: monoclonal antibodiesdrug conjugated https://www.youtube.com/watch?v=ACxmsMmo9hU
- 38. 38 Production of vaccines Typesof Vaccine Whole-Agent Vaccine: Contains whole, non virulent microorganisms Subunit vaccine: Contains some part or product of microrganisms that can produce an immune response Problems These microrganisms can back- mutate to a virulent form Allergic reactions (e.g. against egg substrate used to grow viruses) They can be produced with biotechnology, they are safer than attenuated vaccines because they can’t reproduce and they contain little or no extra material (no allergic reactions)
- 39. 39 Production of recombinantvaccines
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- 41. Gene Therapy Gene therapyis an experimental technique for treating disease by altering the patient's genetic material. Most often, gene therapy works by introducing a healthy copy of a defective gene into the patient's cells. Researchers must find safe, efficient vehicles--called vectors--to carry the modified DNA to targeted cells. There are several vectors already in use, including chemical solutions, synthetic fat molecules, and viruses that have been modified so that they are harmless.
- 42. Gene Therapy Gene therapymay be classified into the two following types, only one of which has been used in humans: Somatic gene therapy As the name suggests, in somatic gene therapy, the therapeutic genes are transferred into the somatic cells (non sex- cells), or body, of a patient. Any modifications and effects will be restricted to the individual patient only, and will not be inherited by the patient's offspring or later generations Germline gene therapy In germline gene therapy, germ cells (sperm or eggs) are modified by the introduction of functional genes, which are integrated into their genomes. Germ cells will combine to form a zygote which will divide to produce all the other cells in an organism and therefore if a germ cell is genetically modified then all the cells in the organism will contain the modified gene. This would allow the therapy to be heritable and passed on to later generations technical and ethical problems=eugenics
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- 44. Gene Therapy totreat SCID A form of SCID is caused by a deficiency of an enzyme (adenosine deaminase or ADA), which is necessary for lymphocytes to develop.
- 45. http://learn.genetics.utah.edu/content/genetherapy/gtdoctor/ Space Doctor Welcome tothe Extraterrestrial Gene Therapy Lab! As Space Doctor, you'll use your gene therapy knowledge to treat three ailing aliens who come into your clinic. A bit of fun…with gene therapy
- 46. Stem cells https://www.youtube.com/watch?v=2-3J6JGN-_Y ESL VIDEO:A Stem Cell Story Quick Quiz
- 47. UNIT 3 • Proand cons of biotechnology
- 50. PRO: advantages ofGMOs Insect Resistance Some GMO foods have been modified to make them more resistant to insect pests. The University of California in San Diego reports that a toxic bacterium can be added to crops to make them insect repellent, yet safe for human use. This can reduce the amount of pesticide chemicals used on the plants, thus potentially reducing exposure to pesticides.
- 51. PRO: advantages ofGMOs More Nutritious Foods The Food and Agricultural Organization of the United Nations reports that some GMO foods have been engineered to be more nutritious in terms of mineral or vitamin content. Not only does this help you get the nutrients you need, it can also play a significant role in battling malnutrition in the developing world. The United Nations advises that vitamin A-enhanced rice is helping to reduce global vitamin A deficiencies. Golden rice
- 52. PRO: advantages ofGMOs More Nutritious Foods Genetically-modified purple tomatoes : Their dark pigment is intended to give tomatoes the same potential health benefits as fruit such as blueberries.
- 53. PRO: advantages ofGMOs More resistant plant •Cold tolerance An antifreeze gene from cold water fish has been introduced into plants such as tobacco and potato. With this antifreeze gene, these plants are able to tolerate cold temperatures that normally would kill unmodified seedlings. •Drought tolerance/salinity tolerance Creating plants that can withstand long periods of drought or high salt content in soil and groundwater will help people to grow crops in formerly inhospitable places
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- 55. 55 Edible vaccines Plants enginereedto contain the vaccine can be grown locally, in the area where vaccination in required, overcoming the logistic and economic problems of transporting prepared vaccines over long distances. Most immportantly, edible vaccines do not require syringes, saving money and eliminating the risk of infection from contamined needles. PRO: advantages of GMOs
- 56. 56 Bioremediation PRO: advantages ofGMOs GM bacteria can clean up oil contamination repidly and effectly, but can you think about the effect caused by the uncontrolled spread of these GM bacteria?
- 58. consumers are theguinea pigs?
- 60. CONS: problems withGMOs Allergic Reactions GMO foods can present significant allergy risks to people. Genetic modification often mixes or adds proteins that weren't indigenous to the original plant or animal, causing new allergic reactions in the human body. In some cases, proteins from an organism that you're allergic to may be added to an organism that you weren't originally allergic to, prompting the same allergic reaction experienced from the first organism.
- 61. CONS: problems withGMOs Possible «escape» of transgene from crops to other species Gene Transfer A constant risk of GMO foods is that the modified genes of the organisms may escape into the wild. Brown University warns that herbicide-resistant genes from commercial crops may cross into the wild weed population, thus creating "superweeds" that are impossible to kill with herbicides More than ten resistant weed species have appeared in the U.S
- 62. CONS: problems withGMOs Gene Transfer A related risk is that the escape of genetically enhanced animals and vegetation can create new super-organisms that can out-compete natural animal and plant populations to drive certain species into extinction.
- 63. CONS: problems withGMOs some crop are genetically modified to resist herbicides, this encourages extensive use of herbicides. Another negative impact is the possibility that increased us of an herbicide will select for weeds with naturally mutations that make them resistant to that herbicide (such as glycofosfate)
- 64. CONS: problems withGMOs Economic concerns Bringing a GM food to market is a lengthy and costly process, and of course agri-biotech companies wish to ensure a profitable return on their investment. Many new plant genetic engineering technologies and GM plants have been patented, and patent infringement is a big concern of agribusiness. Yet consumer advocates are worried that patenting these new plant varieties will raise the price of seeds so high that small farmers and third world countries will not be able to afford seeds for GM crops, thus widening the gap between the wealthy and the poor
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- 66. CONS: problems withGMOs Ethical issues See-Through Frog. GMO Bulls GM animals may suffer poor health and reduced life span Bald Chickens
Editor's Notes
- #4 Biotechnology began when humans began to plant their own crops, domesticate animals, ferment juice into wine, make cheese, and leaven bread These mice are glowing because scientists inserted a gene found in certain bioluminescent jellyfish into their DNA
- #5 Fermented foods, Curd, Cheese and Butter, Organic Acids, Alcoholic Beverages, Industrial solvents, Curing of Beverages,Enzymes, Dextrin, Vitamins, Antibiotics, Tissue Culture, Vaccines, Steroids and old biotechnology is also used in sewage treatment,biogas generation and development of better strains of biofertilizers.
- #8 Corn cob
- #20 This plasmid carries two useful genes, ampR, conferring resistance to the antibiotic ampicillin and lacZ, encoding the enzyme beta-galactosidase which catalyzes the hydrolysis of sugar. Plate out the transformed bacteria on a solid nutrient medium containing ampicillin and a sugar called X-gal. Only bacteria that have the ampicillin-resistance plasmid will grow.The X-gal in the medium is used to identify plasmids that carry foreign DNA. Bacteria with plasmids lacking foreign DNA stain blue when beta-galactosidase hydrolyzes X-gal. Bacteria with plasmids containing foreign DNA are white because they lack beta-galactosidase.
- #23 Figure 7.3 Selection for Recombinant DNA. Selectable marker (reporter) genes are used by biologists to select for bacteria that have taken up a plasmid. In a typical experiment, most of the bacteria will not take up any DNA. Of those that do, only a small fraction will take up recombinant DNA.
- #24 Insulin is a hormone made by the pancreas that allows your body to use sugar (glucose) from carbohydrates in the food that you eat for energy or to store glucose for future use. Insulin helps keeps your blood sugar level from getting too high (hyperglycemia) or too low (hypoglycemia)
- #31 Figure 7.10 Tissue Plasminogen Activator. TPA is a naturally occurring human protein that dissolves blood clots. It is used to treat patients suffering from blood clotting in heart attacks or strokes, and is manufactured using recombinant DNA technology.