Biotechnology

1. Biotechnology Dolly and surrogate Mom Embryonic stem cells and gene therapy Genetically modified rice.

2. Fourteen month-old genetically engineered (“biotech”) salmon (left) and standard salmon (right). Biotechnology Biotechnology is the engineering of organisms for useful purposes. Usually involves the creation of hybrid genes and their introduction into organisms in which some or all of the gene is not normally present.

3. Animal cloning Genetically modified foods Gene cloning for pharmaceutical production DNA fingerprinting The promise and perhaps perils of embryonic stem cells Biotechnology We’ll examine:

4. Animal Cloning Dolly and her surrogate mother 1997

5. Why Clone Animals? Five genetically identical cloned pigs. To answer questions of basic biology For pharmaceutical production. For herd improvement. To satisfy our desires (e.g. pet cloning).

6. Is Animal Cloning Ethical? The first cloned horse and her surrogate mother/genetic twin. As with many important questions, the answer is beyond the scope of science.

7. Video • Researchers isolated a somatic cell from an adult female sheep • Transferred the nucleus from that cell to an egg cell from which the nucleus had been removed • The egg cell, with its new nucleus, was behaving just like a freshly fertilized zygote • It developed into an embryo, which was implanted into a surrogate mother and carried to term. The Biotechnology of Reproductive Cloning

8. Saved by Cloning? Article Some are firm believers while many view these approaches to be more of a stunt.

9. (Science (2002) 295:1443) Carbon Copy– the First Cloned Pet

10. The Next Step? Attempts at human cloning are viewed very unfavorably in the scientific community.

11. Recombinant DNA, Gene Cloning, and Pharmaceutical Production DNA can be cut at specific sequences using restriction enzymes. This creates DNA fragments useful for gene cloning.

12. The enzyme EcoRI cutting DNA at its recognition sequence Restriction Enzymes are Enzymes That Cut DNA Only at Particular Sequences Restriction enzyme animation Different restriction enzymes have different recognition sequences. This makes it possible to create a wide variety of different gene fragments.

13. Palindrome: word that is the same forward and backwords Examples: racecar dad mom 5'- G A A T T C -3‘ 3'- C T T A A G -5'

14. DNA Cut by a Restriction Enzyme Can be Joined Together in New Ways by Ligase These are recombinant DNA and they often are made of DNA from different organisms.

15. Plasmids are Used to Replicate a Recombinant DNA Plasmids are small circles of DNA found in bacteria. Plasmids replicate independently of the bacterial chromosome. Pieces of foreign DNA can be added within a plasmid to create a recombinant plasmid. Replication often produces 50-100 copies of a recombinant plasmid in each cell.

16. Harnessing the Power of Recombinant DNA Technology – Human Insulin Production by Bacteria

17. Human Insulin Production by Bacteria and cut with a restriction enzyme 6) join the plasmid and human fragment

18. Human Insulin Production by Bacteria Mix the recombinant plasmid with bacteria. Screening bacterial cells to learn which contain the human insulin gene is the hard part.

19. Route to the Production by Bacteria of Human Insulin One cell with the recombinant plasmid A fermentor used to grow recombinant bacteria. This is the step when gene cloning takes place. The single recombinant plasmid replicates within a cell. Then the single cell with many recombinant plasmids produces trillions of like cells with recombinant plasmid – and the human insulin gene.

20. Route to the Production by Bacteria of Human Insulin The final steps are to collect the bacteria, break open the cells, and purify the insulin protein expressed from the recombinant human insulin gene.

21. Route to the Production by Bacteria of Human Insulin Overview of gene cloning. Cloning animation

22. Transgenics Transgenic organism: organism genetically engineered by inserting a gene from another organism Transgenic animals, plants, bacteria research, medical, and agricultural purposes

23. Transgenic Bacteria Insulin, growth hormones, and substances that dissolve blood clots Transgenic bacteria slow formation of ice crystals on crops to prevent from crop damage, clean up oil spills, and decompose garbage

24. Transgenic Plants Genetically engineered to have more resistance to insect or viral pests Herbicide and insecticide resistant soybeans, corn, cotton and canola Peanuts and soybeans that do not cause allergic reactions Rice plants with increased iron and vitamins to decrease malnutrition

25. Transgenic Animals Transgenic livestock produced to improve food supply and human health Transgenic goats engineered to secrete a protein called antithrombin III which prevents human blood from forming clots during surgery

27. Pharming These goats contain the human gene for a clot-dissolving protein that is produced in their milk. Pharming is the production of pharmaceuticals in animals engineered to contain a foreign, drug-producing gene.

29. The Guiness Book of Animal Records 1995. "Spider silk is the strongest of all natural and man-made fibres.... It is even stronger than steel: the dragline of a European garden spider (Araneus diadematus), for example, can support a weight of 0.5 g (0.002 oz) without snapping, whereas a steel strand of similar thickness will snap under the strain of just 0.25 g (0.01 oz)." "Spider silk is extremely strong -- it is about five times stronger than steel and twice as strong as Kevlar of the same weight. Spider silk also has the ability to stretch about 30-percent longer than its original length without breaking, which makes it very resilient."

30. The Stem Cell Concept A stem cell is an undifferentiated, dividing cell that gives rise to a daughter cell like itself and a daughter cell that becomes a specialized cell type.

31. The Promise and Possible Perils of Stem Cells

32. Stem Cells are Found in the Adult, but the Most Promising Types of Stem Cells for Therapy are Embryonic Stem Cells

33. The Inner Cell Mass is the Source of Embryonic Stem Cells The embryo is destroyed by separating it into individual cells for the collection of ICM cells.

34. Some Thorny Ethical Questions Are these masses of cells a human? Is it ethical to harvest embryonic stem cells from the “extra” embryos created during in vitro fertilization?

35. DNA, the Law, and Many Other Applications – The Technology of DNA Fingerprinting A DNA fingerprint used in a murder case. The defendant stated that the blood on his clothing was his. What are we looking at? How was it produced?

36. DNA Fingerprinting Basics Different individuals carry different alleles. Most alleles useful for DNA fingerprinting differ on the basis of the number of repetitive DNA sequences they contain.

37. DNA Fingerprinting Basics If DNA is cut with a restriction enzyme that recognizes sites on either side of the region that varies, DNA fragments of different sizes will be produced. DNA samples treated with ethidium bromide are inserted into wells in a piece of agar (gel) When a charge is applied, DNA which is negative will travel to the positive end of the gel Smaller pieces will travel the furthest Bands are visualized under UV light

38. A Site With Three Alleles Useful for DNA Fingerprinting DNA fragments of different size will be produced by a restriction enzyme that cuts at the points shown by the arrows.

39. The DNA Fragments Are Separated on the Basis of Size The technique is called gel electrophoresis. The pattern of DNA bands is compared between each sample loaded on the gel. Gel electrophoresis animation

40. Possible Patterns for a Single “Gene” With Three Alleles In a standard DNA fingerprint, about a dozen sites are analyzed, with each site having many possible alleles.

41. A DNA Fingerprint When many genes are analyzed, each with many different alleles, the chance that two patterns match by coincidence is vanishingly small. DNA detective animation HGP fingerprinting page

42. SLT 3/8/05 DNA and the Law Some applications of DNA fingerprinting in the justice system.

43. Restriction Enzymes and Gel Electrophoresis http://arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/gels/virgel.html Biotechnology Australia site - Games

44. Genetically Modified Foods Many of our crops in the US are genetically modified. Should they be?

45. GM Crops are Here Today Source: Pew Initiative on Food and Biotechnology, August 2004.

46. Methods for Plant Genetic Engineering are Well-Developed and Similar to Those for Animals • Restriction enzyme cut plasmid • Insert gene of interest • Introduce to plant cells

47. Golden rice (yellow) with standard rice (white). Golden Rice is Modified to be Provide a Dietary Source of Vitamin A Worldwide, 7% of children suffer vitamin A deficiency, many of them living in regions in which rice is a staple of the diet.

48. Genetically Modified Crops Genetically Modified Cotton (contains a bacterial gene for pest resistance) Standard Cotton

49. GMOs, Especially Outside the US, Are a Divisive Issue Protesters at the 2000 Montreal World Trade Summit European sentiment

50. Current Concerns by Scientists Focus on Environmental, Not Health, Effects of GM Crops The jury’s still out on the magnitude of GM crop’s ecological impact, but the question is debated seriously.