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DNA The Genetic Material (adapted from K. Foglia)

DNA The Genetic Material (adapted from K. Foglia). Scientific History. The march to understanding that DNA is the genetic material T.H. Morgan (1908) Frederick Griffith (1928) Avery, McCarty & MacLeod (1944) Erwin Chargaff (1947) Hershey & Chase (1952) Watson & Crick (1953)

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DNA The Genetic Material (adapted from K. Foglia)

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  1. DNA The Genetic Material (adapted from K. Foglia)

  2. Scientific History • The march to understanding that DNA is the genetic material • T.H. Morgan (1908) • Frederick Griffith (1928) • Avery, McCarty & MacLeod (1944) • Erwin Chargaff (1947) • Hershey & Chase (1952) • Watson & Crick (1953) • Meselson & Stahl (1958)

  3. 1908 | 1933 Chromosomes related to phenotype • T.H. Morgan • working with Drosophila • fruit flies • associated phenotype with specific chromosome • white-eyed male had specific X chromosome

  4. 1908 | 1933 Genes are on chromosomes • Morgan’s conclusions • genes are on chromosomes • but is it the protein or the DNA of the chromosomes that are the genes? • initially proteins were thought to be genetic material… Why? What’s so impressiveabout proteins?!

  5. 1928 The “Transforming Principle” • Frederick Griffith • Streptococcus pneumonia bacteria • was working to find cure for pneumonia • harmless live bacteria (“rough”) mixed with heat-killed pathogenic bacteria (“smooth”) causes fatal disease in mice • a substance passed from dead bacteria to live bacteria to change their phenotype • “Transforming Principle”

  6. The “Transforming Principle” mix heat-killed pathogenic & non-pathogenic bacteria live pathogenic strain of bacteria live non-pathogenic strain of bacteria heat-killed pathogenicbacteria A. B. D. C. mice die mice live mice live mice die Transformation=change in phenotype something in heat-killed bacteria could still transmit disease-causing properties

  7. 1944 DNA is the “Transforming Principle” • Avery, McCarty & MacLeod • purified both DNA & proteins separately from Streptococcus pneumonia bacteria • which will transform non-pathogenic bacteria? • injected protein into bacteria • no effect • injected DNA into bacteria • transformed harmless bacteria into virulent bacteria What’s the conclusion? mice die

  8. 1944 | ??!! Avery, McCarty & MacLeod • Conclusion • first experimental evidence that DNA was the genetic material Oswald Avery Maclyn McCarty Colin MacLeod

  9. 1952 | 1969 Hershey Confirmation of DNA • Hershey & Chase • classic “blender” experiment • worked with bacteriophage • viruses that infect bacteria • grew phage viruses in 2 media, radioactively labeled with either • 35S in their proteins • 32P in their DNA • infected bacteria with labeled phages Why useSulfurvs.Phosphorus?

  10. Protein coat labeled with 35S DNA labeled with 32P Hershey & Chase T2 bacteriophages are labeled with radioactive isotopes S vs. P bacteriophages infect bacterial cells bacterial cells are agitated to remove viral protein coats Which radioactive marker is found inside the cell? Which molecule carries viral genetic info? 32P radioactivity foundin the bacterial cells 35S radioactivity found in the medium

  11. Blender experiment • Radioactive phage & bacteria in blender • 35S phage (for protein detection) • radioactive proteins stayed in supernatant • therefore viral protein did NOT enter bacteria • 32P phage (for DNA detection) • radioactive DNA stayed in pellet • therefore viral DNA did enter bacteria • Confirmed DNA is “transforming factor” Taaa-Daaa!

  12. 1952 | 1969 Hershey Hershey & Chase Martha Chase Alfred Hershey

  13. There are four kinds of nitrogen bases, so there are four kinds of nucleotides... Purines Pyrimidines Adenine Guanine Cytosine Thymine Nucleotide Phosphate group Deoxyribose

  14. 1947 Chargaff • DNA composition: “Chargaff’s rules” • varies from species to species • all 4 bases not in equal quantity • bases present in characteristic ratio • humans: A = 30.9% T = 29.4% G = 19.9% C = 19.8% RulesA = T C = G That’s interesting!What do you notice?

  15. Chargaff’s basepairing rule The number of A=T G=C Source of DNA A T G C Streptococcus 29.8 31.6 20.5 18.0 Yeast 31.3 32.9 18.7 17.1 Herring 27.8 27.5 22.2 22.6 Human 30.9 29.4 19.9 19.8

  16. 1953 | 1962 Structure of DNA • Watson & Crick • developed double helix model of DNA • other leading scientists working on question: • Rosalind Franklin • Maurice Wilkins • Linus Pauling Wilkins Pauling Franklin

  17. 1953 article in Nature Watson and Crick Watson Crick

  18. Rosalind Franklin (1920-1958)

  19. Hydrogen Bonds P D D C G P Cytosine pairs with Guanine...

  20. P Hydrogen Bonds D T A D P Adenine pairs with Thymine.

  21. P P P P D D T T A D D D D D D C C G G G P P P P A D P P D C Hydrogen bonds

  22. P P P P D D T T A D D D D D D C C G G G P P P P Nucleotide A D P P D C

  23. Because of its chemical properties, DNA is shaped like a Double Helix (twisted ladder) Nucleotide Hydrogen bonds Sugar-phosphate backbone Key Adenine (A) Thymine (T) Cytosine (C) Guanine (G)

  24. Hydrogen bonds Formed between (hydrogen ion and oxygen ion) hold these pairs together, forming the “zipper” between the two strands.

  25. The sugar and phosphate form the backbone of the molecule while the bases are down the center. There are two similar strands (evidences obtained from Rosaline Franklin, 1952)

  26. DNA replication- to duplicate another strand of DNA strand from one DNA template. Happens during S phase (interphase) of the cell cycle. Original strand New strand DNA polymerase Growth DNA polymerase Growth Replication fork Replication fork Nitrogen bases Original strand New strand

  27. But how is DNA copied? • Replication of DNA • base pairing suggests that it will allow each side to serve as a template for a new strand “It has not escaped our notice that the specific pairing we have postulated immediately suggests a possible copying mechanism for the genetic material.” —Watson & Crick

  28. Can you designa nifty experimentto verify? Models of DNA Replication • Alternative models • become experimental predictions conservative semiconservative dispersive P 1 2

  29. Meselson & Stahl Matthew Meselson Franklin Stahl Franklin Stahl Matthew Meselson

  30. Scientific History • March to understanding that DNA is the genetic material • T.H. Morgan (1908) • genes are on chromosomes • Frederick Griffith (1928) • a transforming factor can change phenotype • Avery, McCarty & MacLeod (1944) • transforming factor is DNA • Erwin Chargaff (1947) • Chargaff rules: A = T, C = G • Hershey & Chase (1952) • confirmation that DNA is genetic material • Watson & Crick (1953) • determined double helix structure of DNA • Meselson & Stahl (1958) • semi-conservative replication

  31. The “Central Dogma” • Flow of genetic information in a cell transcription translation DNA RNA protein replication

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