1. Fig. 12-00

2. Fig. 12-01

3. Fig. 12-02

4. Fig. 12-03

5. Fig. 12-04

6. Fig. 12-05

7. Fig. 12-06

8. Fig. 12-07 Plasmids Bacterial chromosome Remnant of bacterium Colorized TEM

9. Fig. 12-08-1 Plasmid Bacterial cell Isolate plasmids.

10. Fig. 12-08-2 Plasmid Bacterial cell Isolate plasmids. DNA Isolate DNA. Cell containing the gene of interest

11. Fig. 12-08-3 Plasmid Bacterial cell Isolate plasmids. DNA Isolate DNA. DNA fragments from cell Cut both DNAs with same enzyme. Gene of interest Other genes Cell containing the gene of interest

12. Fig. 12-08-4 Plasmid Bacterial cell Isolate plasmids. Gene of interest Recombinant DNA plasmids DNA Isolate DNA. DNA fragments from cell Cut both DNAs with same enzyme. Gene of interest Other genes Mix the DNAs and join them together. Cell containing the gene of interest

13. Fig. 12-08-5 Plasmid Bacterial cell Isolate plasmids. Recombinant bacteria Gene of interest Recombinant DNA plasmids Bacteria take up recombinant plasmids. DNA Isolate DNA. DNA fragments from cell Cut both DNAs with same enzyme. Gene of interest Other genes Mix the DNAs and join them together. Cell containing the gene of interest

14. Fig. 12-08-6 Plasmid Bacterial cell Isolate plasmids. Clone the bacteria. Recombinant bacteria Bacterial clone Gene of interest Recombinant DNA plasmids Bacteria take up recombinant plasmids. DNA Isolate DNA. DNA fragments from cell Cut both DNAs with same enzyme. Gene of interest Other genes Mix the DNAs and join them together. Cell containing the gene of interest

15. Fig. 12-08-7 Plasmid Bacterial cell Isolate plasmids. Find the clone with the gene of interest. Clone the bacteria. Recombinant bacteria Bacterial clone Gene of interest Recombinant DNA plasmids Bacteria take up recombinant plasmids. DNA Isolate DNA. DNA fragments from cell Cut both DNAs with same enzyme. Gene of interest Other genes Mix the DNAs and join them together. Cell containing the gene of interest

16. Fig. 12-08-8 Plasmid Bacterial cell Isolate plasmids. Some uses of genes Gene for pest resistance Gene for toxic-cleanup bacteria Genes may be inserted into other organisms. Find the clone with the gene of interest. The gene and protein of interest are isolated from the bacteria. Clone the bacteria. Recombinant bacteria Bacterial clone Gene of interest Recombinant DNA plasmids Bacteria take up recombinant plasmids. Harvested proteins may be used directly. Some uses of proteins Protein for “stone-washing” jeans DNA Cell containing the gene of interest Protein for dissolving clots Isolate DNA. DNA fragments from cell Cut both DNAs with same enzyme. Gene of interest Other genes Mix the DNAs and join them together.

17. Fig. 12-09-1 Recognition sequence for a restriction enzyme Restriction enzyme Sticky end Sticky end DNA A restriction enzyme cuts the DNA into fragments.

18. Fig. 12-09-2 Recognition sequence for a restriction enzyme Restriction enzyme Sticky end Sticky end DNA A DNA fragment is added from another source. A restriction enzyme cuts the DNA into fragments.

19. Fig. 12-09-3 Recognition sequence for a restriction enzyme Restriction enzyme Sticky end Sticky end DNA A DNA fragment is added from another source. A restriction enzyme cuts the DNA into fragments. Fragments stick together by base pairing.

20. Fig. 12-09-4 Recognition sequence for a restriction enzyme Restriction enzyme Sticky end Sticky end DNA ligase Recombinant DNA molecule A DNA fragment is added from another source. A restriction enzyme cuts the DNA into fragments. Fragments stick together by base pairing. DNA ligase joins the fragments into strands.

21. Fig. 12-10 Radioactive probe (single-stranded DNA) Single-stranded DNA Mix with single-stranded DNA from various bacterial clones Base pairing indicates the gene of interest

22. Fig. 12-11-1 Cell nucleus DNA of eukaryotic gene Test tube Transcription Exon Intron Exon Intron

23. Fig. 12-11-2 Cell nucleus DNA of eukaryotic gene RNA transcript mRNA Test tube Transcription Introns removed and exons spliced together Exon Intron Exon Intron

24. Fig. 12-11-3 Cell nucleus DNA of eukaryotic gene RNA transcript mRNA Test tube Reverse transcriptase Transcription Introns removed and exons spliced together Isolation of mRNA from cell and addition of reverse transcriptase Exon Intron Exon Intron

25. Fig. 12-11-4 Cell nucleus DNA of eukaryotic gene RNA transcript mRNA Test tube Reverse transcriptase cDNA strand being synthesized Transcription Introns removed and exons spliced together Isolation of mRNA from cell and addition of reverse transcriptase Synthesis of cDNA strand Exon Intron Exon Intron

26. Fig. 12-11-5 Cell nucleus DNA of eukaryotic gene RNA transcript mRNA Test tube cDNA of gene without introns Reverse transcriptase cDNA strand being synthesized Transcription Introns removed and exons spliced together Isolation of mRNA from cell and addition of reverse transcriptase Synthesis of cDNA strand Synthesis of second DNA strand by DNA polymerase Exon Intron Exon Intron

27. Fig. 12-12

28. Fig. 12-13-1 DNA isolated Crime scene Suspect 1Suspect 2

29. Fig. 12-13-2 DNA isolated DNA amplified Crime scene Suspect 1Suspect 2

30. Fig. 12-13-3 DNA isolated DNA amplified DNA compared Crime scene Suspect 1Suspect 2

31. Fig. 12-14

32. Fig. 12-14a

33. Fig. 12-14b

34. Fig. 12-15 Initial DNA segment Number of DNA molecules 1 248

35. Fig. 12-16 Crime scene DNA Suspect’s DNA Same number of short tandem repeats Different numbers of short tandem repeats STR site 1STR site 2 AGAT GATA

36. Fig. 12-17-1 Mixture of DNA fragments of different sizes Power source Gel

37. Fig. 12-17-2 Mixture of DNA fragments of different sizes Power source Gel

38. Fig. 12-17-3 Mixture of DNA fragments of different sizes Power source Gel Completed gel Band of longest (slowest) fragments Band of shortest (fastest) fragments

39. Fig. 12-18 Amplified crime scene DNA Amplified suspect’s DNA Longer fragments Shorter fragments

40. Fig. 12-19 Crime scene DNA Suspect’s DNA Fragment w Fragment x Fragment y Longer fragments Shorter fragments Fragment z Fragment y Crime scene DNA Suspect’s DNA Cut Restriction enzymes added x w y y z

41. Fig. 12-20

42. Fig. 12-21 Anthrax spore Envelope containing anthrax spores

43. Fig. 12-22-1 Chromosome

44. Fig. 12-22-2 Chromosome Chop up with restriction enzyme DNA fragments

45. Fig. 12-22-3 Chromosome Chop up with restriction enzyme Sequence fragments DNA fragments

46. Fig. 12-22-4 Chromosome Chop up with restriction enzyme Sequence fragments DNA fragments Align fragments

47. Fig. 12-22-5 Chromosome Chop up with restriction enzyme Sequence fragments DNA fragments Align fragments Reassemble full sequence

48. Fig. 12-22a

49. Fig. 12-23

50. Fig. 12-24-1 Normal human gene isolated and cloned Healthy person

51. Fig. 12-24-2 Normal human gene isolated and cloned Normal human gene inserted into virus Healthy person Harmless virus (vector) Virus containing normal human gene

52. Fig. 12-24-3 Normal human gene isolated and cloned Normal human gene inserted into virus Virus injected into patient with abnormal gene Healthy person Harmless virus (vector) Virus containing normal human gene Bone marrow Bone of person with disease

53. Fig. 12-25

54. Fig. 12-26

55. Fig. 12-27

56. Fig. 12-28

57. Fig. 12-T01

58. Fig. 12-UN01 DNA isolated from two sources and cut by same restriction enzyme Gene of interest (could be obtained from a library or synthesized) Recombinant DNA Plasmid (vector) Transgenic organisms Useful products

59. Fig. 12-UN02 Crime sceneSuspect 1Suspect 2 DNA Polymerase chain reaction (PCR) amplifies STR sites Longer DNA fragments Shorter DNA fragments DNA fragments compared by gel electrophoresis Gel

60. Fig. 12-UN03 Normal human gene Virus Bone marrow Normal human gene is transcribed and translated in patient, potentially curing genetic disease permanently