Mutations Germ Mutations Affect only the reproductive cells (egg & sperm)

1. Mutations Germ Mutations Affect only the reproductive cells (egg & sperm) Mutation passed on to offspring Somatic Mutation Affect only body cells Mutation NOT passed on to offspring

2. Point Mutations (1 nucleotide is affected) Substitution (May change in ONE amino acid) Insertion (Frameshift mutation) Deletion (Frameshift mutation) Frameshift mutations DNA sequence is “shifted” left or right Downstream codons are changed…drastic change in the protein. **We ALL have some potentially dangerous mutations

3. Polyploidy More than two sets of chromosomes. Occurs because of a problem in chromosome separation in Meiosis, call Nondisjunction.

4. Normal Meiosis should produce haploid gametes. All gametes are haploid; they each have one copy of each chromosome. When two gametes are used for fertilization, each new cell should be have two copies of each chromosome and be diploid.

5. Nondisjunction in Meiosis I:

6. Nondisjunction in Meiosis II:

7. Selective Breeding Pick best traits and cross them (potato) 1. Inbreeding Cross those with similarities (Dogs) Can result in an increase in genetic defects in inbred population. (cheetahs, dogs) 2. Hybridization Cross those with different characteristics Results in hardier offspring (Corn) 3. Mutagens Create mutations using chemicals, UV light, etc. Selection for best or most desirable traits (seedless grapes)

8. Genetic Engineering (Techniques that directly affect the DNA) DNA Extraction Polymerase Chain Reaction (PCR) Restriction Enzymes (RE) cut DNA DNA Fingerprints Transformation (Recombinant DNA) Gene Therapy Cloning (Somatic Cell Nuclear Transfer) DNA Sequencing

9. DNA Extraction (Get DNA out of the cell.) “Mash up cells” (blend) Add Water Add Salt Add baking powder Add detergent Breaks down cell membrane Dissolves lipids and proteins Filter out solution Add solution to cold isopropyl alcohol DNA is not soluble in alcohol See stringy white DNA

10. Polymerase Chain Reaction (PCR) Add Primer Heat Add A, C, G, T Add DNA Polymerase Repeat 20-30 cycles Millions of copies of the desired portion of DNA!

11. Figure 13-8 PCR Section 13-2 DNA polymerase adds complementary strand DNA heated to separate strands DNA fragment to be copied PCRcycles 1 DNAcopies 1 3 4 4 8 5 etc. 16 etc. 2 2 Go to Section:

12. Restriction Enzymes (RE) Cut DNA at specific base sequences CTTAAG GAATTC Example of EcoRI RE AACTTAAGTTTTTCTTAAGCCC TTGAATTCAAAAAGAATTCGGG RE’s leave fragments that have “sticky ends”. These ends have unpaired bases. AAC T T A A GCCC TTG AAT T CGGG

13. Restriction Enzymes Section 13-2 Recognition sequences DNA sequence Restriction enzyme EcoRI cuts the DNA into fragments. Sticky end Go to Section:

14. Restriction Enzymes Section 13-2 Recognition sequences DNA sequence Restriction enzyme EcoRI cuts the DNA into fragments. Sticky end Go to Section:

15. Figure 13-6 Gel Electrophoresis Section 13-2 Power source DNA plus restriction enzyme Longer fragments Shorter fragments Gel Mixture of DNA fragments Go to Section:

16. Original Well B D C A DNA is negatively charged DNA moves away from negative (-) terminal; and towards the positive (+) terminal Shortest pieces move through porous gel fastest. Longest pieces move more slowly. Thickest bands represent most common fragment size. A is longest fragment. D is shortest fragment. D is also the most common fragment length. DNA Gel Electrophoresis

17. PCR Copies a segment of DNA Blood DNA is Extracted DNA is broken Into fragments by Restriction Enzymes Radioactive Probes Label DNA Gel Electrophoresis (separates based on size) DNA Fingerprint is read.

18. DNA --- Person 1 TTGTGGCCCCCCAATTGTT 1 CTTAGAAAGGAGGGGAAGT 2 ATGAGATTTTTTTTTAGGC 3 ACACACAAGAGATATAGAG 4 AAAAATTGTGGTGTAGAGC 5 CCCCGAAAAAAAAAAAACA 6 CACACAAGATAGATGTGTG 7 TGCGCGCGGGGGGGAATAA 8 CAGTGTTGTATTAATTTAT 9 AGAAAATAAGATATATGGG 10

19. DNA --- Person 2 TTGTGGCCCCCCAATTGTT 1 ATTAGAGGGGAGGGGAAGT 2 ATGAGATTTTTCTTTATGC 3 ACACACATGAGATATAAAG 4 AACAATTGTGGTGTAGAGC 5 CCCCGAAAACCCCAAAACA 6 CACAAAAGATAGATGTGTG 7 TGAGCGCGGGGGGGAATCT 8 CAGTGTTGTATTAACCTAT 9 AGAAAATTTGATATATGGG 10

20. DNA --- Person 1 TTGTGGCCCCCCAATTGTTCTTAGAAAGGAGGGGAAGTATGAGATTTTTTTTTAGGCACACACAAGAGATATAGAGAAAAATTGTGGTGTAGAGCCCCCGAAAAAAAAAAAACACACACAAGATAGATGTGTGTGCGCGCGGGGGGGAATAA CAGTGTTGTATTAATTTATAGAAAATAAGATATATGGG

21. DNA --- Person 1 TTGTGGCCCCCCAATTGTTCTTAGAAAG GAGGGGAAG TATGAGATTTTTTTTTAGGCACACACAAG AGATATAGAGAAAAATTGTGGTGTAGAGCCCCCGAAAAAAAAAAAACACACACAAG ATAGATGTGTGTGCGCGCGGGGGGGAATAACAGTGTTGTATTAATTTATAGAAAATAAG ATATATGGG

22. DNA --- Person 2 TTGTGGCCCCCCAATTGTTATTAGAGGGGAGGGGAAGT ATGAGATTTTTCTTTATGCACACACATGAGATATAAAGAACAATTGTGGTGTAGAGCCCCCGAAAACCCCAAAACACACAAAAGATAGATGTGTGTGAGCGCGGGGGGGAATCT CAGTGTTGTATTAACCTATAGAAAATTTGATATATGGG

23. DNA --- Person 2 TTGTGGCCCCCCAATTGTTATTAGAGGGGAGGGGAAG TATGAGATTTTTCTTTATGCACACACATGAGATATAAAG AACAATTGTGGTGTAGAGCCCCCGAAAACCCCAAAACACACAAAAG ATAGATGTGTGTGAGCGCGGGGGGGAATCTCAGTGTTGTATTAACCTATAGAAAATTTGATATATGGG

24. - - - - DNA Person #2 ATAGATGTGTGTGAGCGCGGGGGGGAATCTCAGTGTTGTATTAACCTATAGAAAATTTGATATATGGG ATAGATGTGTGTGCGCGCGGGGGGGAATAACAGTGTTGTATTAATTTATAGAAAATAAG AGATATAGAGAAAAATTGTGGTGTAGAGCCCCCGAAAAAAAAAAAACACACACAAG AACAATTGTGGTGTAGAGCCCCCGAAAACCCCAAAACACACAAAAG TATGAGATTTTTCTTTATGCACACACATGAGATATAAAG TTGTGGCCCCCCAATTGTTATTAGAGGGGAGGGGAAG TTGTGGCCCCCCAATTGTTCTTAGAAAG TATGAGATTTTTTTTTAGGCACACACAAG DNA Person #1 GAGGGGAAG ATATATGGG + + + +

25. Blood from victim A well 1 Blood from victim B well 2 Semen collected on victim A well 3 Semen collected on victim B well 4 Blood from suspect X well 5 Blood from suspect Y well 6 Blood from suspect Z well 7 Results from the Restriction Enzyme (RE) Digest: 1 2 3 4 5 6 7

26. Mother’s blood well 1 Child’s blood well 2 Possible Father 1 well 3 Possible Father 2 well 4 Possible Father 3 well 5 Who is the father? Results from the Restriction Enzyme Digest: Mother Child Father #1 Father #2 Father #3

27. TRANSFORMATION EcoRI RE site “Sticky Ends” Add RE INSULIN + “Recombinant DNA” Insulin gene inside bacteria. *Bacteria produce Human INSULIN!!! INSULIN EcoRI RE sites

28. Figure 13-9 Making Recombinant DNA Section 13-3 Gene for human growth hormone Recombinant DNA Gene for human growth hormone DNA recombination Human Cell Sticky ends DNA insertion Bacterial Cell Bacterial chromosome Bacterial cell for containing gene for human growth hormone Plasmid Go to Section:

29. Transgenic tobacco plant with a gene from a firefly.

30. Gene Therapy Replace a defective gene with a good copy. Put “good” gene into a virus. Have virus infect your cells. Virus puts DNA (with good gene) into YOUR cells. Not YET common. Best treatment so far has been treatment of SCIDS. (Bubble Boy Syndrome). Promising treatment for a form of muscular dystrophy and a form of blindness known as Leber’s congenital amaurosis (LCA).

31. Figure 13-10 Plant Cell Transformation Section 13-3 Agrobacterium tumefaciens Gene to be transferred Cellular DNA Inside plant cell, Agrobacterium inserts part of its DNA into host cell chromosome Recombinant plasmid Plant cell colonies Transformed bacteria introduce plasmids into plant cells Complete plant is generated from transformed cell Go to Section:

32. Flowchart Section 13-4 Cloning A body cell is taken from a donor animal. An egg cell is taken from a donor animal. The nucleus is removed from the egg. The body cell and egg are fused by electric shock. The fused cell begins dividing, becoming an embryo. The embryo is implanted into the uterus of a foster mother. The embryo develops into a cloned animal. Go to Section:

33. Figure 13-13 Cloning of the First Mammal Section 13-4 A donor cell is taken from a sheep’s udder. Donor Nucleus These two cells are fused using an electric shock. Fused Cell Egg Cell The nucleus of the egg cell is removed. An egg cell is taken from an adult female sheep. The fused cell begins dividing normally. Embryo Cloned Lamb The embryo is placed in the uterus of a foster mother. The embryo develops normally into a lamb—Dolly Foster Mother Go to Section:

34. Single strand of DNA Fluorescent dye Strand broken after A Power source Strand broken after C Strand broken after G Strand broken after T Gel Figure 13-7 DNA Sequencing Section 13-2 Go to Section:

35. The Future of DNA Sequencing? • Pacific Biosciences SMRT DNA Sequencing • DNA Sequencing in under an hour for $100? • http://www.pacificbiosciences.com/video_lg.html

36. The Power of RNAi (RNA Interference) • RNAi NOVA Video 15 min • http://www.pbs.org/wgbh/nova/sciencenow/3210/02.html

37. Can we bring back extinct organisms? • 60 Minutes "Resurrecting the Extinct" (12 min) • http://www.cbsnews.com/video/watch/?id=6078982n&tag=contentMain;cbsCarousel

38. Quick Video from HHMI • HHMI 2002 Holiday Lecture Series • http://media.hhmi.org/hl/02Lect1.html • 41:46  Clip 26 through 50:00 (8 minutes) • 1000 Smell receptors