Chapter 7 DNA replication

Chapter 7 DNA replication - Chapter 7 DNA Replication...

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Unformatted text preview: Chapter 7 DNA Replication Suggested Problems: 2, 3, 4, 5, 7, 8, 13, 19, 21 Outline: A. Semiconservative replication B. Historical experiments 1. Meselson-Stahl 2. Cairns C. Replication 1. Leading strand/Lagging strand 2. The replisome 3. Replication initiation a. Prokaryotes b. Eukaryotes 4. Replication termination. D. PCR E. Dideoxy sequencing Possible ways DNA could be replicated Meselson-Stahl experiment proved semiconservative model Labeled parental E. coli DNA with a heavy isotope of nitrogen ( 15 N). Newly synthesized strands with 14 N (instead of 15 N) are shown in black. Separate 14 N and 15 N by centrifugation John Cairns showed that E. coli chromosomes replicate from an origin, using a replication fork labeled DNA with radioactive hydrogen during or after replication, he fixed the cells and covered them with photograhic emulsion sites with radioactivity show black dots Reaction catalyzed by DNA polymerase III Energy for the reaction comes from breaking the high-energy phosphate bond on the 5 carbon. Semiconservative DNA replication Function of DNA Polymerase III DNA polymerase III copies DNA in a 5 3 direction only DNA polymerase III also has 3 5 exonuclease activity (involved in repair of mismatches) DNA polymerase III cannot start cold---it can extend a chain of nucleotides, but it cannot start one. Steps in lagging strand synthesis: Other enzymes: primase: adds RNA primer DNA pol I : removes RNA, fills...
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Chapter 7 DNA replication - Chapter 7 DNA Replication...

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