History of DNA Scientific history The journey to understanding that DNA is our genetic material T.H. Morgan (1908) Frederick Griffith (1928) Avery, McCarty,

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Presentation transcript:

History of DNA

Scientific history The journey to understanding that DNA is our genetic material T.H. Morgan (1908) Frederick Griffith (1928) Avery, McCarty, & MacLeod (1944) Erwin Chargaff (1947) Hersey & Chase (1952) Rosalind Franklin (1952) Watson & Crick (1953) Meselson & Stahl (1958)

T.H. Morgan Chromosomes are related to phenotype Worked with Drosophila Fruit flies Associated phenotype with specific chromosomes White-eyed male had a specific X chromosome

T.H. Morgan Conclusion Genes are on chromosomes But is it the protein or the DNA of the chromosome that codes for the gene? Initially proteins were thought to contain genetic information … why? What ’ s so impressive about proteins?!

Frederick Griffith Streptococcus pneumonia bacteria Was working to find a 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 ”

Frederick Griffith – Transforming Principle Transformation = change in phenotype something in heat-killed bacteria could still transmit disease-causing properties live pathogenic strain of bacteria live non-pathogenic strain of bacteria mice diemice live heat-killed pathogenic bacteria mix heat-killed pathogenic & non-pathogenic bacteria mice livemice die A.B. C. D.

Avery, McCarty, & MacLeod Purified both DNA and 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 mice die What ’ s the conclusion?

Avery, McCarty, & MacLeod Conclusion DNA is the “Transforming Principle” First experimental evidence that DNA housed the genetic material

Hershey & Chase Confirmation of DNA Classic “blender” experiment Worked with bacteriophage Viruses that infect bacteria Grew phage virus in 2 media, both radioactively labeled 35 S in the proteins 32 P in the DNA Infected bacteria with labeled phages Why use S ulfur vs. P hosphorus?

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

Hershey & Chase Blender experiment Radioactive phage & bacteria in blender 35S phage Radioactive proteins stayed in supernatant Therefore viral protein did NOT enter bacteria 32P phage Radioactive DNA stayed in pellet Therefore viral DNA did enter bacteria Confirmed DNA is transforming factor Taaa-Daaa!

Chargaff DNA composition: “Chargaff’s rules” Varies from species to species All 4 bases not in equal quantity Bases present in characteristic ratios Humans A = 30.9% T = 29.4% G = 19.9% C = 19.8% That ’ s interesting! What do you notice? Rules A = T C = G

Rosalind Franklin 1 st to “photograph” DNA molecule X-ray crystallography Raymond Gosling : lab assistant; actually took the picture Maurice Wilkins: 1 st to attempt technique; set-up lab Franklin used

Watson & Crick Developed double helix model of DNA 1953 article in Nature

Meselson & Stahl How is DNA copied? 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

Meselson & Stahl Alternate models presented Become experimental predictions conservativesemiconservativedispersive 1 2 P Can you design a nifty experiment to verify?

Meselson & Stahl Label “parent” nucleotides in DNA strands with heavy nitrogen = 15 N Label new nucleotides with lighter nitrogen isotope = 14 N “The Most Beautiful Experiment in Biology” Make predictions … 15 N parent strands 15 N/ 15 N parentreplication

Meselson & Stahl 1st round of replication conservative 15 N/ 15 N 14 N/ 14 N semi- conservative 15 N/ 14 N dispersive 15 N/ 14 N conservative 15 N/ 15 N 14 N/ 14 N semi- conservative 15 N/ 14 N dispersive 15 N/ 14 N 2nd round of replication 14 N/ 14 N 15 N parent strands 15 N/ 15 N 1 Experiment predictions Experiment results 2

Scientific history The journey to understanding that DNA is our 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’s rules: A = T, C = G Hersey & Chase (1952) Confirmation that DNA is genetic material Rosalind Franklin (1952) Photographed DNA molecule Watson & Crick (1953) Determined double helix structure of DNA Meselson & Stahl (1958) Semi-conservative replication

Any Questions??