1. PowerLecture: Chapter 16
Studying and Manipulating Genomes

2. Genetic Changes
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3. Discovery of Restriction Enzymes
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4. Specificity of Cuts
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5. Making Recombinant DNA5’ G
A A T T C 3’
C T T A A G
one DNA fragment
another DNA fragment 5’ G
A A T T C 3’ 3’
C T T A A G 5’

6. Making Recombinant DNA
nick 5’ G
A A T T C 3’ 3’
C T T A A G 5’
nick
DNA ligase action G
A A T T C
C T T A A G

7. enzyme recognition siteA G
cut fragments 3’ 5’
enzyme recognition site G C T A
another DNA fragment A T C 3’ 5’ G
one DNA fragment
DNA ligase action
nick G C T A 3’ 5’ G C T A 3’ 5’
Stepped Art
Fig. 16-2, p.244

8. Using Plasmids ________________________________________________

9. Plasmids
Fig. 16-3a, p.244

10. Plasmids
Fig. 16-3b, p.244

11. Restriction enzyme cuts molecule of chromosomal DNA or cDNA
fragments with
sticky ends
Foreign DNA, plasmid
DNA, and modification
enzymes are mixed
recombinant plasmids
containing foreign DNA
Same enzyme cuts same sequence in plasmid DNA
plasmid DNA
with sticky ends
host cells containing recombinant plasmids
Stepped Art
Fig. 16-4, p.245

12. Making cDNA (complimentary DNA)
Fig. 16-5, p.245

13. Gene Libraries ___________________________________________

14. Using a Probe to Find a Gene
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15. Use of a Probe Colonies on plate Cells adhere to filter
Cells are lysed;
DNA sticks
to filter
Probe is
added
Location where probe binds forms
dark spot on film, indicates colony with gene

16. Amplifying DNA
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17. Polymerase Chain Reaction
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18. Polymerase Chain Reaction
Double-stranded
DNA to copy
DNA heated to
90°– 94°C
Primers added to
base-pair with
ends
Mixture cooled;
base-pairing of
primers and ends
of DNA strands
DNA polymerases
assemble new
DNA strands
Stepped Art
Fig. 16-6, p. 256

19. Polymerase Chain Reaction
Mixture heated again; makes all DNA fragments unwind
Mixture cooled; base-pairing between primers and ends of single DNA strands
DNA polymerase action again doubles number of identical DNA fragments
Stepped Art
Fig. 16-6, p. 256

20. Recording the Sequence
p.248

21. Gel Electrophoresis _____________________________
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22. Reaction Mixture
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23. Nucleotides for Sequencing
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24. Reactions Proceed
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25. Recording the Sequence
T C C A T G G A C C
T C C A T G G A C
Recording the Sequence
T C C A T G G A
T C C A T G G
T C C A T G
T C C A T
T C C A
electrophoresis
gel
T C C
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T C
one of the many fragments of
DNA migrating
through the gel T
one of the DNA fragments
passing through a laser beam
after moving through the gel
T C C A T G G A C C A

26. Recording the Sequence
Fig. 16-8b, p.248

27. DNA Fingerprints ________________________________
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28. Tandem Repeats
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29. RFLPs ___________________________________
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30. Gel Electrophoresis
Fig. 16-9a, p.249

31. Gel Electrophoresis
Fig. 16-9b, p.249

32. Analyzing DNA Fingerprints
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33. Genome Sequencing
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34. Genome Sequencing
Fig a, p.250

35. Genomics
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36. DNA Chips
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37. DNA Chips
Fig , p.251

38. Genetic Engineering
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39. Engineered Proteins _________________________________________________

40. Cleaning Up the Environment
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41. Can Genetically Engineered Bacteria “Escape”?
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42. p.252

43. Engineered Plants
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44. Engineered Plants
Fig a, p.253

45. Engineered Plants
Fig b, p.253

46. The Ti plasmid
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plant cell
foreign gene
in plasmid

47. The Ti plasmid
b The bacterium infects a plant and transfers the Ti plasmid into it.
a A bacterial cell contains a Ti plasmid (purple) that has a foreign gene (blue).
e Young plants with a fluorescent
gene product.
c The plant cell divides.
d Transgenic plants.
Fig , p.253

48. First Engineered Mammals
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49. Transgenic Mice
Fig , p.254

50. Designer Cattle ____________________________________________________

51. Genetically Modified Animals Featherless (created by traditional cross breeding)
Fig a, p.254

52. Genetically Modified Animals Mira a transgenic goat that makes a human anticlotting factor
Fig b, p.254

53. Genetically Modified Animals
Fig c, p.254

54. Xenotransplantation
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55. Safety __________________
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56. Safety
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57. The Human Genome Initiative
Goal - Map the entire human genome
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58. Using Human Genes
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59. Ethical Issues
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