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The History Of DNA. Quick Review. Chromosomes are made up of DNA and Proteins . Genes are a segment of the DNA molecule that carries the instructions for producing a specific traits. 1952). DNA or Proteins?.
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The History Of DNA Honors Biology 2006-2007
Honors Biology 2006-2007
Quick Review • Chromosomes are made up of DNA and Proteins. • Genes are a segment of the DNA molecule that carries the instructions for producing a specific traits. Honors Biology 2006-2007
1952) DNA or Proteins? • Prior to the 1950’s, scientists could not decide whether DNA or protein was the material responsible for heredity. • 3 experiments would lead to the answer: • 1. Frederick Griffith • 2. Avery, McCarty & MacLeod • 3. Hershey & Chase Honors Biology 2006-2007
1928 Griffith’s Experiment • Frederick Griffith was working to find a cure for Streptococcus pneumonia (bacteria) • Test Subjects: lab mice • He used different strains (types) of bacterium in this experiment. Honors Biology 2006-2007
Strains of Bacteria Used • S Bacteria = Disease Causing (pathogenic) • Have protective outer coat that “hides” the bacteria from the body’s immune defenses. • R Bacteria = Do not cause disease (non-pathogenic) • Lacked the protective coat and thus the immune system can identify and destroy them. Honors Biology 2006-2007
The Experimental Trials 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? something in heat-killed bacteria could still transmit disease-causing properties Honors Biology 2006-2007
Griffith’s “Transforming Factor” • Why did the mice die that were given a mixture of heat-killed pathogenic bacteria and live non-pathogenic bacteria? • A substance had been passed from the dead bacteria to live bacteria = “Transforming Factor” Honors Biology 2006-2007
Griffith’s Results • After examining blood samples, he found that somehow the uncoated bacteria had TRANSFORMED into bacteria with coats. • Transformation is a change in the genetic makeup caused when cells take up foreign genetic material. BUT, what was it? DNA or Protein?? Honors Biology 2006-2007
What Next? • Whatever the molecule, it had to have several properties in order to fit the bill: • It had to be duplicated whenever a cell divided, so it could be passed on unchanged. • It had to be in the form of an informational code • It had to be (mostly) stable and resistant to change Honors Biology 2006-2007
Avery, McCarty & MacLeod 1944 Oswald Avery Colin MacLeod Maclyn McCarty Honors Biology 2006-2007
DNA causes Transformation • Avery, McCarty & MacLeod • purified both DNA & proteins from Streptococcus pneumonia bacteria • which will transform non-pathogenic bacteria? • Experiment 1: • injected protein into bacteria = no effect • injected DNA into bacteria = transformation! • Experiment 2: • added protein destroying enzymes to bacteria = transformation! • added DNA destroying enzymes to bacteria = no effect Honors Biology 2006-2007
Hershey & Chase Martha Chase Alfred Hershey Honors Biology 2006-2007
1952 | 1969 Confirmation of DNA • Hershey & Chase • “Blender” experiment using bacteriophages • viruses that infect bacteria • Half of the phages had their proteins tagged with radioactive sulfur (35S) • The other half had their DNA tagged with radioactive phosphorus (32P) • All phage were then allowed to infect bacteria! Honors Biology 2006-2007
Honors Biology 2006-2007
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 liquid Honors Biology 2006-2007
Taaa-Daaa! Blender experiment Results • Radioactive phage & bacteria in blender • 35S phage • radioactive proteins were in liquid • therefore protein did NOT enter bacteria • 32P phage • radioactive DNA was in pellet • therefore DNA did enter bacteria Honors Biology 2006-2007
Hershey & Chase’s Conclusion • Hershey & Chase Confirmed DNA is “transforming factor” • Proved that DNA AND NOT protein is the hereditary material in cells (i.e., genes are made up of DNA) Honors Biology 2006-2007
Wilkin’s and Franklin’s Photographs • In 1952, Maurice Wilkins and Rosalind Franklin, developed high quality X-ray diffraction photographs of strands of DNA. • These photos were key pieces of data used to determine the helical structure of DNA molecules. Honors Biology 2006-2007
1953 article in Nature Watson and Crick Honors Biology 2006-2007
The Structure of DNA Honors Biology 2006-2007
Deoxyribonucleic Acid • DNA is the molecule of heredity. • DNA (Deoxyribonucleic Acid) is a double-stranded nucleic acid that determines an organism’s traits by controlling the production of proteins. • DNA holds the information for life! • Why do we study DNA?? Honors Biology 2006-2007
DNA’s Size • DNA is an extremely long molecule • DNA from one of your cells would measure 6 feet tall! • All of the DNA in your body could stretch to the sun and back 400 times! The earth is 93 million miles from the sun! • 5 million strands of DNA can fit through the head of one needle! Honors Biology 2006-2007
DNA’s Helical Structure • Watson & Crick discovered DNA is made up of 2 chains of nucleotides joined together by their nitrogen bases. • The 2 strands are twisted together, forming a Double-Helix. • Like a spiral staircase, twisted ladder, or zipper Honors Biology 2006-2007
Subunits of DNA • When we group the deoxyribose sugar, phosphate group and nitrogen base together into a subunit it is called a Nucleotide. • One strand of DNA has many millions of nucleotides. nucleotide Honors Biology 2006-2007
O -P O O O C C Nucleotides Continued • Sugar = Deoxyribose. • The Phosphate Group = Phosphorus atom surrounded by 4 oxygen atoms. • The Nitrogen Bases are carbon ring structures that contain one or more atoms of nitrogen. Nitrogenous base Phosphate O C C C Deoxyribose O Honors Biology 2006-2007
What is DNA made of? phosphate • The backbone of the molecule is alternating phosphate groups and deoxyribose sugars. • The “rungs” of the DNA “ladder” are nitrogenousbases. deoxyribose bases Honors Biology 2006-2007
Four nitrogenous bases • DNA has four different nitrogenous bases: • Adenine (A) • Thymine (T) • Cytosine (C) • Guanine (G) Honors Biology 2006-2007
N N C N C C C N N C Two Classes of DNA Bases • Pyrimidines are single ring bases. • Thymine • Cytosine (Pyrimidins have a “y”) • Purines are double ring bases. • Adenine • Guanine N N C O C C N C Honors Biology 2006-2007
Chargaff’s Rule • The amount of Adenine is always equal to the amount of Thymine. • Adenine and Thymine have a 1:1 ratio • The amount of Guanine is always equal to the amount of Cytosine. • Guanine and Cytosine have a 1:1 ratio Honors Biology 2006-2007
Base pairing in DNA • Pairing • Adenine and Thymine always join together • Two Hydrogen Bonds • Cytosine and Guanine always join together • 3 Hydrogen Bonds • Adenine-Thymine and Guanine-Cytosine are called Complimentary Base Pairs. Honors Biology 2006-2007
hydrogen bonds How do the strands stick? • Hydrogen Bondsform between the nitrogenous bases. • Weak, but there are millions and millions of them in a single molecule of DNA. • Remember hydrogen bonds can form between hydrogen and any electronegative atom! Honors Biology 2006-2007
Nucleotide Sequence • Nucleotide Sequence is extremely important. • Differences in organisms are due to the difference in sequence of the nucleotides along a strand of DNA. • Nucleotide sequence forms the unique genetic information of an organism. Honors Biology 2006-2007
DNA Replication Honors Biology 2006-2007
Copying DNA • Each cell in an organism has an exact copy of the DNA that was in the fertilized egg (Mitosis) • The DNA in the chromosome is copied in a process called DNA Replication. • If DNA wasn’t copied before cell division, new cells would only have half the DNA of their parents. Honors Biology 2006-2007
DNA Replication • Base pairing allows each strand to serve as a pattern (template) for a new strand • Semi-Conservatice Model • 2 molecules are formed. • Each has an original strand and one new strand. Honors Biology 2006-2007
let’s meetthe team… DNA Replication • Large team of enzymes coordinates replication: • DNA Helicase - unwinds the DNA so that it can be copied • DNA Polymerase(s) - Brings in the nucleotide building blocks to make the complimentary strand AND “proofreads” the finished strands. • DNA Ligase - Acts as “glue” cementing new pieces of DNA together to form a continuous strand. Honors Biology 2006-2007
1st step: Unwinding • Unwind & unzip DNA • helicase enzyme • unwinds DNA helix • stabilized bysingle-stranded binding proteins • The areas where the double helix separate are called replication forks. single-stranded binding proteins Honors Biology 2006-2007
direction of replication Replication fork Honors Biology 2006-2007
Now, that’s a compliment! 2nd step - Adding Bases • DNA Polymerases bring in new nucleotides that match up to template strands Honors Biology 2006-2007
Results of DNA Replication • DNA Replication results in the formation of 2 DNA molecules, each identical to the original DNA molecule. • Each new molecule has one strand from the original DNA molecule and one new strand. • Semi-conservative! Honors Biology 2006-2007
Checking for Errors • Sometimes errors occur when the wrong nucleotide is added to the new strand. • Luckily, DNA Polymerase has a “proofreading function.” • DNA Polymerase can backtrack and remove incorrect nucleotides. Honors Biology 2006-2007
Fast & accurate! • It takes E. coli <1 hour to copy 5 million base pairs in its single chromosome • divide to form 2 identical daughter cells • Human cell copies its 6 billion bases & divide into daughter cells in only few hours • remarkably accurate • only ~1 error per 100 million bases • ~30 errors per cell cycle Honors Biology 2006-2007