DNA Synthesis Eukaryotes DNA unwinds at a replication origin Two replication forks form Bidirectional synthesis of DNA Much more DNA; complexed with proteins

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  • DNA Synthesis Eukaryotes DNA unwinds at a replication origin Two replication forks form Bidirectional synthesis of DNA Much more DNA; complexed with proteins Eukaryotes have much more DNA Yeast 250-400 replicons Mammalian cells up to 25,000 Solution: Multiple replication origins Yeast 4X that of E. coli Drosophila 100X that of E. coli
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  • DNA Synthesis in Eukaryotes Pre-initiation replication complex: ORC Cdt1 Cdc6 MCM
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  • Enzyme FunctionProkaryotesEukaryotes Helicase Primase Polymerase Proofreading Sliding clamp Prevent ss closure DnaB DnaG Pol III sliding clamp SSBPs MCM Primase s.u. of -polymerase Pol and PCNA (Proliferating Cell Nuclear Antigen) RPA (Replication Protein A) Comparison of some replication components
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  • A Problem and a Solution
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  • Telomere Formation
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  • A model for t loop formation
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  • Reproductive cloning Biotechnology/Cloning
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  • Reproductive cloning
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  • Therapeutic cloning
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  • http://www.rndsystems.com/dam_public/6515.jpg Therapeutic cloning
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  • First Cloning Experiment Cohen et al., 1973 Recombinant DNA
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  • Cloning Vectors: pBR322 plasmid Cloning into pBR322
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  • RT-PCR and Cloning