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DNA as Genetic Material DNA vs. Protein At first protein was thought to be genetic material - 20 possible AAs in Protein, 4 nucleotides in DNA Griffith’s experiment - Fredrick Griffith found that nonpathogenic bacteria could be made pathogenic by incubating with heatkilled pathogenic bacteria - bacteria were “transformed” Avery-MacLeod-McCarty experiment identify transforming substance in Griffith’s experiment - Isolated DNA, RNA, and Protein from heat-killed pathogenic bacteria - tested which would transform live nonpathogenic bacteria - transformation occurred w/DNA only Hershey-Chase experiment (blender experiment) Used T2 bacteriophage (virus that infects bacteria) to determine if protein or DNA is genetic material - DNA contains P and not S - protein contains S and not P - radioactive S and P were used to label protein and DNA - radioactive P was found in E. coli not S when bacteriophage infected Opener 12-12-16 Explain how in the Hershey-Chase experiment they were able to determine if it were DNA or protein that is transferred from bacteriophages to bacteria during infection. 3-D structure of DNA Erwin Chargaff identified that amount of A and T is Equal, and amount of G and C is equal (Chargaff’s Rule) Rosalind Franklin X-ray crystallography Rosalind Franklin made X ray images of DNA crystals - saw distinct diffraction pattern - James Watson determined that the diffraction pattern was of a helical molecule made of 2 strands James Watson and Francis Crick - Worked out DNA base pairing, explains Chargaff’s rule - Determined that DNA strands are antiparallel - finalized 3-d structure DNA replication Process that occurs in S Interphase Semiconservative nature of replication - Replication begins with 1 double stranded DNA and makes 2 double-stranded daughter DNA molecules - each daughter DNA contains 1 of original parent DNA strands Replication process DNA unwinding (helicase) Priming (primase) Synthesis (DNA polymerase) Process begins at origins of replication (OOR) Unwinding DNA Helicase enzyme breaks hydrogen bond between base pairs Opens up DNA for replication enzymes to have access Priming DNA polymerase cannot initiate synthesis with DNA nucleotides -RNA primer is used to get DNA polymerase started -Primase adds a few RNA nucleotides, base paired to the parent strand Synthesis DNA polymerase matches nucleotides to the template strand by base pair rules - Adds nucleotides to 3’ end of primer - catalyzes reaction that links nucleotides together along the backbone of new DNA strand Leading and lagging strands DNA polymerase can only add nucleotides to 3’ end of growing strand For each daughter DNA being synthesized there is leading strand and a lagging strand Leading strand grows from OOR in 3’ direction Lagging strand is filled in discontinuously on the 5’ end of the strand Lagging strand synthesis Numerous RNA primers used to initiate fragments (called Okazaki fragments) DNA polymerase I replaces RNA primer with DNA nucleotides Ligase used to link backbone of fragments Telomeres Eukaryotic chromosomes are linear, they have an ending Telomeres are special sequences at the end of chromosomes Each replication event results in a shortening of Telomeres In gametes cells telomerase enzyme lengthens the telomeres