Chapter 16. Molecular Basis of Inheritance
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1 Chapter 16 Molecular Basis of Inheritance
2 DNA genetic material Chromosomes composed of DNA + protein
3 Evidence for DNA 1. Griffith 1928 Streptococcus pneumonia S strain death Inject heat-killed bacteria what happens?
4 Inject live R strain mouse lives R is not pathogenic!
5 Inject killed S + live R strain --- Conclusion: Killed S strain transformed R strain How? What? Griffiths experiment
6 2. evidence: The transforming factor is DNA Hershey and Chase 1952
7 Bacteriophage (virus) Infects bacteria many new viruses made, cell dies Virus composed of protein + DNA
8 Radioactivity in the lab 35 S-met labels viral proteins 32 P labels DNA
9 1. Infect bacteria with 35 S-phage (radioactive protein) 2. Strip off phage from bacterial membrane 3. No radioactivity found in cells Conclusion: viral protein does not enter E. coli
10 1. Infect bacteria with 32 P-phage (radioactive DNA) 2. Strip off phage from bacterial membrane P found in cells Conclusion: Viral DNA enters E. coli, is the genetic material
11 3. DNA base composition Nucleotide base Guanine Cytosine Thymine Adenine
12 DNA is a polymer of nucleotides
13 Chargaff s rules (1950) [T] = [A] [G] = [C] A certain chromosome is 19% A. What is the % of C? Purines are larger than pyrimidines
14 4. DNA structural model Watson, Crick, Franklin 1953 X-ray crystallography DNA is helical Spacing of bases Width of helix suggested 2 strands
15 DNA double helix Sugar-phosphate backbone Anti parallel strands
16 Bases face inward Hydrogen bonds connect bases
17 A - T (2 bonds) G - C ( 3 bonds)
18 DNA replication: history Can read quotes from W + C s paper in textbook 3 models of DNA replication, 1 correct
19 Original DNA copied to new DNA helix Original DNA broken up and combined in new DNA 1 strand original DNA maintained in new DNA
20 Meselson and Stahl 1950s 1. Label DNA (E. coli) with 15 N in growth media 2. Transfer E. coli to 14 N media for 1 generation (20 min)
21 Results: The density of the DNA is intermediate Cells grown longer 14 N, make lighter DNA
22 What would the DNA density be after 20 more minutes of cell group? 14 N DNA gm/cm 3 15 N DNA1.724 gm/cm 3
23 DNA replication is semi-conservative
24 DNA replication: mechanism (E. coli) Human genome = 3 X 10 9 bp DNA (X2) 46 chromosomes E. coli genome = 4 X 10 6 bp DNA 1 circular chromosome 1 origin of replication (ori)
25 Ori Specific sequence of nucleotides Replication proteins attach to ori Forms a replication bubble Two strands of DNA open
26 Replication fork in both directions
27 Proteins in DNA replication Table DNA polymerase (enzyme) Adds nucleotides 5 3 direction only
28 2. Helicase (enzyme) unwinds double helix 3. Single stranded binding protein (SSB) binds to DNA strands to stabilize them
29 4. Topoisomerase (enzyme) breaks, rejoins DNA to relieve physical stress 5. Primase synthesizes a primer
30 Leading strand is 3 5 Lagging strand is 5 3 Each strand is a template for new DNA
31 DNA replication leading strand: steps 1. Primase (enzyme) synthesizes primer complementary to leading strand primer is ~10 bases
32 2. DNA polymerase (pol III) synthesizes new strand 5 3 G, A, T, C nucleotides complementary to template strand 500 nuc/sec Continuous elongation until end of chromosome
33 DNA Synthesis steps: lagging strand 1. Primase makes RNA primer 2. DNA adds nucleotides to primer in 5 3 direction only
34 3. DNA pol III detaches Okazaki fragment ~ 1, 000 nucleotides long
35 4. Another primer added, another Okazaki fragment formed Many primers needed
36 5. Gaps between primers filled in 6. Ligase enzyme bonds fragments
37 DNA replication Fig
38 Telomeres, the protective ends
39 Linear DNA has telomeres No genes Repetitive DNA TTAGGG up to 1000 times 5'...TTAGGG TTAGGG TTAGGG TTAGGG TTAGGG TTAGGG..3 3'...AATCCC AATCCC AATCCC AATCCC AATCCC AATCCC..5' Human chromosomes capped by telomeres
40 Chromosomes shorten with each cell division
41 When telomeres are too short cell senescence (irreversible) ~ 125 cell divisions (humans) life span? Mouse fibroblasts in culture
42 Cells that do not divide often Example: heart muscle Telomeres do not shorten with age
43 Telomeres shorten ~100 bp each time cell divides Lagging strand problem
44 Embryonic cells, some wbc, stem cells, cancer cells express telomerase White blood cell cervical cancer cell embryo
45 Telomerase enzyme synthesizes telomeres Animation garland
46 Telomerase action and the end replication problem Telomerase has both enzyme and RNA
47 Dolly s DNA was 5 years old at the time of cloning Her telomeres were only 80% the length of other 1 year olds
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