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Biology - Genetic Engineering

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  • 1. Genetic Engineering Biology - Edgar
  • 2. Fig. 20-3-1 Restriction site DNA Sticky end Restriction enzyme cuts sugar-phosphate backbones. 5  3  3  5  1
  • 3. Fig. 20-3-2 Restriction site DNA Sticky end Restriction enzyme cuts sugar-phosphate backbones. 5  3  3  5  1 DNA fragment added from another molecule cut by same enzyme. Base pairing occurs. 2 One possible combination
  • 4. Fig. 20-3-3 Restriction site DNA Sticky end Restriction enzyme cuts sugar-phosphate backbones. 5  3  3  5  1 One possible combination Recombinant DNA molecule DNA ligase seals strands. 3 DNA fragment added from another molecule cut by same enzyme. Base pairing occurs. 2
  • 5. Fig. 20-9a Mixture of DNA mol- ecules of different sizes Power source Longer molecules Shorter molecules Gel Anode Cathode TECHNIQUE 1 2 Power source – + + –
  • 6.  
  • 7.  
  • 8.  
  • 9. Fig. 20-10 Normal allele Sickle-cell allele Large fragment (b) Electrophoresis of restriction fragments from normal and sickle-cell alleles 201 bp 175 bp 376 bp (a) Dde I restriction sites in normal and sickle-cell alleles of  -globin gene Normal  -globin allele Sickle-cell mutant  -globin allele Dde I Large fragment Large fragment 376 bp 201 bp 175 bp Dde I Dde I Dde I Dde I Dde I Dde I
  • 10.  
  • 11. Genetic Engineering & Cloning
  • 12. Herbert Boyer and Stanley Cohen
  • 13.  
  • 14.  
  • 15.  
  • 16. Isolate DNA
  • 17. Restriction Enzymes
  • 18.  
  • 19. Cut DNA
  • 20.  
  • 21.  
  • 22.  
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  • 27.  
  • 28.  
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  • 32.  
  • 33.  
  • 34.  
  • 35. Recombinant DNA
  • 36.  
  • 37.  
  • 38.  
  • 39.  
  • 40.  
  • 41.  
  • 42. Polymerase Chain Reaction
  • 43. Polymerase Chain Reaction
    • In Scientific American , Mullis summarized the accomplishment: "Beginning with a single molecule of the genetic material DNA, the PCR can generate 100 billion similar molecules in an afternoon. The reaction is easy to execute. It requires no more than a test tube, a few simple reagents, and a source of heat." [9]
    http:// www.youtube.com/watch?v =xd4De47ldYs
  • 44.  
  • 45.  
  • 46.  
  • 47.  
  • 48. PCR
    • The PCR Song
    • The Concept.
  • 49. GMO Lab
  • 50. Step 1
    • Identify protein that can improve crop.
      • Bt crops: produce delta-endotoxin which is lethal to corn borers.
  • 51. Step 2
    • Isolate gene – Clone
  • 52. Step 3
    • Engineer gene so that crop will “read” it.
      • Removing interons, and add promoter Cauliflower mosaic virus promoter
      • CaMV 35S
  • 53. Step 3
      • Add terminator: NOS terminator from Agrobacterium tumefaciens
  • 54. Get the Gene into the Crop
    • Agrobacterium tumefaciens Crown Gall Disease
  • 55. Fig. 12-8a-1 Agrobacterium tumefaciens DNA containing gene for desired trait Ti plasmid Insertion of gene into plasmid Recombinant Ti plasmid 1 Restriction site
  • 56. Fig. 12-8a-2 Agrobacterium tumefaciens DNA containing gene for desired trait Ti plasmid Insertion of gene into plasmid Recombinant Ti plasmid 1 Restriction site Plant cell Introduction into plant cells 2 DNA carrying new gene
  • 57. Fig. 12-8a-3 Agrobacterium tumefaciens DNA containing gene for desired trait Ti plasmid Insertion of gene into plasmid Recombinant Ti plasmid 1 Restriction site Plant cell Introduction into plant cells 2 DNA carrying new gene Regeneration of plant 3 Plant with new trait
  • 58. 2 Primers
    • GMO Primers: Duplex PCR:
      • CaMV 35S Promoter
      • NOS Terminator
    • Plant Primers: Photosystem II Chloroplast Gene
  • 59. Gel Electrophoresis
    • 1: non-GMO food with plant primers.
    • 2: non-GMO food with GMO primers
    • 3: Test food with plant primers
    • 4: Test food with GMO primers.
    • 5: GMO-positive template with plant primers.
    • 6: GMO-positive template with GMO primers.
    • 7: PCR molecular weight ruler.
  • 60. The Questions
    • Is Genetically Modified food safe to eat?
    • Just how radical is this new technology?
    • What are the benefits?
    • What are the risks?
    • Are GM foods sufficiently regulated in the US?
  • 61. GMO
    • PBS Harvest of Fear: GM Fish
  • 62. 11,497 4700 2838 1700 1159 805 Pst I PstI 923 SspI 2586 HpaI 849 1257

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