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n Chapter 16~ The Molecular Basis of Inheritance
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Searching for Genetic Material n Hershey and Chase √ bacteriophages (phages) √ DNA, not protein, is the hereditary material √ Expt: sulfur(S) is in protein, phosphorus (P) is in DNA; only P was found in host cell
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DNA Structure n Chargaff ratio of nucleotide bases (A=T; C=G) n Watson & Crick The Double Helix √ nucleotides: nitrogenous base (thymine, adenine, cytosine, guanine); sugar deoxyribose; phosphate group
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DNA Bonding n Purines: ‘A’ & ‘G’ n Pyrimidines: ‘C’ & ‘T’ (Chargaff rules) n ‘A’ H+ bonds (2) with ‘T’ and ‘C’ H+ bonds (3) with ‘G’ n Van der Waals attractions between the stacked pairs
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DNA Replication n Watson & Crick strands are complementary; nucleotides line up on template according to base pair rules (Watson) n Meselson & Stahl replication is semiconservative; Expt: varying densities of radioactive nitrogen
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DNA Replication: a closer look n Origin of replication (“bubbles”): beginning of replication n Replication fork: ‘Y’-shaped region where new strands of DNA are elongating n Helicase: catalyzes the untwisting of the DNA at the replication fork n DNA polymerase III: catalyzes the elongation of new DNA
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DNA Replication, II n Antiparallel nature: sugar/phosphate backbone runs in opposite directions (Crick); one strand runs 5’ to 3’, while the other runs 3’ to 5’; DNA polymerase only adds nucleotides at the free 3’ end, forming new DNA strands in the 5’ to 3’ direction only
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DNA Replication, IIIReplication, n Leading strand: synthesis toward the replication fork (only in a 5’ to 3’ direction from the 3’ to 5’ master strand) n Lagging strand: synthesis away from the replication fork (Okazaki fragments); joined by DNA ligase (must wait for 3’ end to open; again in a 5’ to 3’ direction) n Initiation: Primer (short RNA sequence~w/primase enzyme), begins the replication process n DNA polymerase I- removes RNA primer and replaces with DNA
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DNA Repair n Mismatch repair: DNA polymerase n Excision repair: Nuclease n Telomere ends: telomerase
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