Division Ave. High School AP Biology
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1 DNA The Genetic Material Scientific History The march to understanding that DNA is the genetic material u T.H. Morgan (1908) u Frederick Griffith (1928) u Avery, McCarty & MacLeod (1944) u Erwin Chargaff (1947) u Hershey & Chase (1952) u Watson & Crick (1953) u Meselson & Stahl (1958) Chromosomes related to phenotype T.H. Morgan u working with Drosophila fruit flies u associated phenotype with specific chromosome white-eyed male had specific X chromosome 1
2 Genes are on chromosomes Morgan s conclusions u genes are on chromosomes u 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 impressive about proteins?! The Transforming Principle Frederick Griffith u Streptococcus pneumonia bacteria was working to find cure for pneumonia u harmless live bacteria ( rough ) mixed with heat-killed pathogenic bacteria ( smooth ) causes fatal disease in mice u a substance passed from dead bacteria to live bacteria to change their phenotype Transforming Principle 1928 The Transforming Principle mix heat-killed pathogenic & live pathogenic strain of bacteria live non-pathogenic strain of bacteria heat-killed pathogenic bacteria non-pathogenic bacteria A. B. C. D. mice die mice live mice live mice die Transformation = change in phenotype something in heat-killed bacteria could still transmit disease-causing properties 2
3 1944 DNA is the Transforming Principle Avery, McCarty & MacLeod u purified both DNA & proteins separately from Streptococcus pneumonia bacteria which will transform non-pathogenic bacteria? u injected protein into bacteria no effect u injected DNA into bacteria transformed harmless bacteria into virulent bacteria mice die What s the conclusion? 1944??!! Avery, McCarty & MacLeod Conclusion u first experimental evidence that DNA was the genetic material Oswald Avery Maclyn McCarty Colin MacLeod Confirmation of DNA Hershey & Chase u classic blender experiment u worked with bacteriophage viruses that infect bacteria u grew phage viruses in 2 media, Why use radioactively labeled with either Sulfur 35 S in their proteins vs. Phosphorus? 32 P in their DNA u infected bacteria with labeled phages Hershey 3
4 Hershey & Chase Protein coat labeled with 35 S T2 bacteriophages are labeled with radioactive isotopes S vs. P DNA labeled with 32 P bacteriophages infect bacterial cells Which radioactive marker is found inside the cell? bacterial cells are agitated to remove viral protein coats Which molecule carries viral genetic info? 35 S radioactivity found in the medium 32 P radioactivity found in the bacterial cells Blender experiment Radioactive phage & bacteria in blender u 35 S phage radioactive proteins stayed in supernatant therefore viral protein did NOT enter bacteria u 32 P phage radioactive DNA stayed in pellet therefore viral DNA did enter bacteria u Confirmed DNA is transforming factor Taaa-Daaa! 4
5 Hershey & Chase Hershey Martha Chase Alfred Hershey 1947 Chargaff DNA composition: Chargaff s rules u varies from species to species u all 4 bases not in equal quantity u bases present in characteristic ratio humans: A = 30.9% T = 29.4% G = 19.9% C = 19.8% Rules A = T C = G That s interesting! What do you notice? Structure of DNA Watson & Crick u developed double helix model of DNA other leading scientists working on question: wrosalind Franklin wmaurice Wilkins wlinus Pauling Franklin Wilkins Pauling 5
6 Division Ave. High School 1953 article in Nature Watson and Crick Watson Crick Rosalind Franklin ( ) But how is DNA copied? Replication of DNA u 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 6
7 Models of DNA Replication Alternative models u become experimental predictions P conservative semiconservative dispersive Semiconservative replication Meselson & Stahl u label parent nucleotides in DNA strands with heavy nitrogen = 15 N u label new nucleotides with lighter isotope = 14 N The Most Beautiful Experiment in Biology Make predictions 15 N/ 15 N parent replication 15 N parent strands Predictions 1st round of replication 8 14 N/ 14 N 15 N/ 15 N 15 N/ 14 N 15 N/ 14 N 2nd round of replication P 1 15 N/ 15 N 2 15 N parent strands conservative 14 N/ 14 N 15 N/ 15 N conservative 4 14 N/ 14 N 15 N/ 14 N semiconservative semiconservative dispersive N/ 14 N dispersive 7
8 Meselson & Stahl Matthew Meselson Franklin Stahl Franklin Stahl Matthew Meselson Scientific History March to understanding that DNA is the genetic material u T.H. Morgan (1908) genes are on chromosomes u Frederick Griffith (1928) a transforming factor can change phenotype u Avery, McCarty & MacLeod (1944) transforming factor is DNA u Erwin Chargaff (1947) Chargaff rules: A = T, C = G u Hershey & Chase (1952) confirmation that DNA is genetic material u Watson & Crick (1953) determined double helix structure of DNA u Meselson & Stahl (1958) semi-conservative replication The Central Dogma Flow of genetic information in a cell transcription DNA RNA translation protein replication 8
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