1. 1 DNA Technology

2. 2 DNA Technology is used for 1. produce pharaceutical products 2. produce herbicide resistant crops 3. treat genetic diseases

3. 3 What is MOLECULAR BIOLOGY? Bio on the molecular level Like GENETIC ENGINEERING -when we alter/manipulate genes For what reasons? –Cure diseases/mutations (GENE THERAPY) –Improve food crops –What else?!

4. 4 1. DNA Extraction Chemical treatmentsChemical treatments (like adding detergents) cause cells and nuclei to burst and remove DNA from cells stickyThe DNA is inherently sticky, and can be pulled out of the mixture “ spooling ”This is called “ spooling ” DNA

5. 5 “ Spooled ” DNA

6. 6 2. Splicing (cutting) DNA Restriction enzymesRestriction enzymes (ex. nuclease) cut DNA at specific sequences manageable fragmentsDivides DNA into manageable fragments

7. 7 Sticky ends -places on DNA where R.E. cut

8. 8 3. Transferring Genes Uses a PLASMID – is an extra ring of DNA –Ring of DNA in a bacterium (it’s not the main chromosome tho)

9. 9 RECOMBINANT PLASMID -changed DNA the plasmid acts like a host for the cloning vector to be placed inside

10. 10 How does the prokaryote know how to express eukaryote genes? AH! YOU MAKE SURE YOU CUT THE TARGET GENE WITH ITS PROMOTER!

11. 11

12. 12 Making insulin

13. 13 TRANSGENIC ORGANISM -host organism that gets rDNA (recombinant DNA)

14. 14 4. Gel Electrophoresis sizeDNA fragments can be separated based on size phosphate groups negativelyThe phosphate groups on outside backbone are negatively charged gelelectricityDNA is placed in a gel and electricity is run through

15. 15 Gel Electrophoresis Negative DNANegative DNA moves toward the positive end Smaller farther and fasterSmaller fragments move farther and faster

16. 16 What cuts DNA into smaller chunks so they can drop out throughout the gel?

17. 17 DNA FINGERPRINTING by Gel Electrophoresis Pattern of bands made up of specific DNA fragments

18. 18 Which suspect was at the crime scene?!

19. 19 5. DNA SEQUENCING Figure out the ORDER of NUCLEOTIDES (bases) Inject DNA with dyes that will bind to specific bases Bases will drop out at certain known “heaviness” spots so you can sequence it!

20. 20 6. PCR - Copying DNA Polymerase Chain ReactionPolymerase Chain Reaction Also called PCR A method of making many quick copies of a piece of DNA

21. 21 Steps in Copying DNA A DNA molecule is placed in a small test tube +DNA polymerase+DNA polymerase +heat to separate into ss+heat to separate into ss +primers (ss matching DNA pieces to start replication)+primers (ss matching DNA pieces to start replication) +nucleotides+nucleotides

22. 22 Copying DNA DNA polymerase adds new bases to the separated strandsThe tube is cooled, and DNA polymerase adds new bases to the separated strands

23. 23 PCR Large amounts of DNA can be made from a small starting sample

24. Dr. Kary Mullis 1993 Dr. Kary Mullis (1944) discovered the PCR procedure, while HIGH on LSD!

25. 25 7. Cloning CloneClone- can produce many genetically identical cells or offspring asexual reproductionproduced by asexual reproduction (mitosis) We can quickly produce organisms with desirable traits

26. 26 Cloning organisms body cellA body cell from one organism egg celland an egg cell from another are fused divides like a normal embryoThe resulting cell divides like a normal embryo

27. What was the first cloned organism? 27

28. 28 Who was the 1 st cloned MAMMAL ? Dolly (born 1996) and her surrogate Mommy

29. Dolly and her baby, Bonnie 29

30. Dolly died in 2003 (put to sleep after seeing lung tumors) 30 Normal sheep lifespan 10-12 yrs

31. We clone animals…(lab and domesticated) 31

32. 2001 We’ve brought back an extinct animal – the bucardo goat (but it died of lung defect) 32

33. List of animal clones 1 Carp 2 Cat 3 Cattle 4 Deer 5 Dog 6 Ferret 7 Frog (tadpole) 8 Fruit flies 9 Gaur 10 Goat 11 Horse 12 Mice 33 13 Mouflon 14 Mule 15 Pig 16 Pyrenean ibex 17 Rabbit 18 Rat 19 Rhesus Monkey 20 Sheep 21 Water Buffalo 22 Wolf 23 Zebrafish

34. 2007 1 st primate stem cells cloned 34

35. 2013 1 st HUMAN stem cells cloned 35

36. 36 CLONING = DE-EXTINCTING ANIMALS MAMMOTH -overhunting

37. 37 GASTRIC-BROODING FROG -habitat loss, pollution, disease? (Australia)

38. 38 PASSENGER PIGEON -overhunting (fed to slaves!)

39. 39 DODO -overhunted by sailors (Mauritius)

40. 40 8. Human Genome Project

41. 41 Human Genome Project 1990 -2003 Research effort to sequence all of our DNA (46 chromosomes) Over 3 billion nucleotides Mapping every gene location (loci) Conducted by scientists around the world

42. 42 Benefits of Human Genome Project Improvements in medical prevention of disease, gene therapies, diagnosis techniques …Improvements in medical prevention of disease, gene therapies, diagnosis techniques … Production of useful protein products for use in medicine, agriculture, bioremediation and pharmaceutical industries.Production of useful protein products for use in medicine, agriculture, bioremediation and pharmaceutical industries. Improved bioinformatics – using computers to help in DNA sequencing …Improved bioinformatics – using computers to help in DNA sequencing …

43. 43 9. Biotechnology - TO create new products from ALTERING GENES in plants and animals for more desirable traits

44. 44 GM FOODS -Genetically Modified

45. 45 Herbicide Resistant Crops Soybeans: Roundup Ready Corn: Roundup Ready, Liberty Link Cotton: BXN, Roundup Ready Canola: Liberty Link, Roundup Ready + CP4 EPSPS = Roundup gene Ready

46. 46 Biotechnology Breakthroughs Rice –Enriched with beta-carotene and iron Bananas –Containing edible hepatitis vaccine Rice –Enriched with beta-carotene and iron Bananas –Containing edible hepatitis vaccine

47. 47 Biotechnology Breakthroughs Potatoes with higher solid content Garlic that lowers cholesterol Potatoes with higher solid content Garlic that lowers cholesterol

48. GM vs Artificial Selection 48

49. 49 Pros Reduced pesticide use Lower energy requirements Cleaner water Less soil erosion More food production! Cheaper! $ Reduced pesticide use Lower energy requirements Cleaner water Less soil erosion More food production! Cheaper! $

50. 50 Cons Not natural Does it cause health issues? Should it be labeled? Not natural Does it cause health issues? Should it be labeled?

51. 51 Corn – insecticide Soy – herbicide Cotton – pesticide Papaya – virus- resistant Rice – Vitamin A Dairy – bovine GH

52. 52 10. Epigenetics – altering genes by messing with OUTSIDE DNA stuff like RNA or HISTONES! Which mouse did we methylate (add CH3 group to its histone proteins that DNA wraps around?

53. 11. iPSC induced Pluripotent Stem Cells -turning BODY cells BACK into STEM cells! (no embryo killing) 53 Transfect with viral vectors But…  Causes cancer bc virus doesn’t know when to stop telling cells to reproduce  the rate at which body cells are reprogrammed into stem cells is LOW! 0.1%! Bc we need to know the EXACT time, balance and levels of expression of gene we want.

54. Stem cells are undifferentiated cells 54