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Lecture 5 DNA replication pt 4

Lecture 5 DNA replication pt 4 - DNA replication Part 4...

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DNA replication Part 4 http://blogs.jobdig.com/wwds/files/2008/08/lobsters_500.jpg 1 Wednesday, September 7, 2011
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Last Time The gamma complex of DNAPIII and the role of the beta clamp “Ring on a String” experiments that demonstrate how the B-Clamp associates with DNA Stewart, J. et al. (2001) results show how the gamma complex associates with the B-clamp. 2 Wednesday, September 7, 2011
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Learning Outcomes 1. Describe the process of replication initiation and termination (in proks). 2. Describe how the origin of replication was identified ( experiment ) and be able to apply this technique to a new problem. 3. Describe mtDNA replication and identify ways it is both similar to and different from E. coli replication. 4. Illustrate how telomerase works. 5. Discuss how mtDNA mutations and telomere mutations are associated with aging phenotypes. By the end of this lecture, you should be able to: 3 Wednesday, September 7, 2011
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DNA replication Pt 4 1. Three phases of prokaryotic DNA replication ( experiment: defining the origin of replication ) 2. Replication of mtDNA 3. Replication in Eukaryotes Image courtesy: http://www.brunel.ac.uk/2206/SHSSC/Telomerase-2.jpg 4 Wednesday, September 7, 2011
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1. Initiation 2. Elongation 3. Termination Three phases of DNA replication 5 Wednesday, September 7, 2011
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How is an origin of replication selected? Primosome assembly at the origin The oriC (Bacterial) Origin Initiation 6 Wednesday, September 7, 2011
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Fig 21.3 Experimental Question: what portion of DNA is crucial for replication to occur? purify Amp gene 7 Wednesday, September 7, 2011
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Fig 21.3 reiterative 8 Wednesday, September 7, 2011
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OriC consensus sequence 245 bp minimal region required for replication initiation in E.coli Fig. 21.4 Comparison with other bacterial genomes shows this region is highly conserved! 9 Wednesday, September 7, 2011
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Fig. 21.4 10 Wednesday, September 7, 2011
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Fig. 21.4 DnaA boxes: 11 Wednesday, September 7, 2011
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+ Hu on the origin + ATP Ready to bind primase (DnaG)! Similar to Fig. 21.5 12 Wednesday, September 7, 2011
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lagging strand leading strand DnaB helicase DnaG primase Interaction of DnaB, DnaG and Pol III From: Lewin, Genes (2000) 13 Wednesday, September 7, 2011
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2. Many copies of dnaA protein bind the four 9-mers within oriC ; DNA wraps around dnaA forming “Initial Complex”.
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