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DNA Synthesis

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 4X that of E. coli •Drosophila 100X that of E. coli. •Solution:

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DNA Synthesis

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  1. 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 4X that of E. coli •Drosophila 100X that of E. coli •Solution: Multiple replication origins •Yeast 250-400 replicons •Mammalian cells up to 25,000

  2. DNA Synthesis in Eukaryotes Pre-initiation replication complex: ORC Cdt1 Cdc6 MCM

  3. Comparison of some replication components

  4. A Problem and a Solution

  5. Telomere Formation

  6. A model for t loop formation

  7. Biotechnology/Cloning Reproductive cloning

  8. Reproductive cloning

  9. Therapeutic cloning

  10. Therapeutic cloning http://www.rndsystems.com/dam_public/6515.jpg

  11. First Cloning Experiment Cohen et al., 1973 “Recombinant DNA”

  12. Cloning into pBR322 Cloning Vectors: pBR322 plasmid

  13. RT-PCR and Cloning

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