mol5 - BIMM100! Lecture 5 ! Chapter 5 - Recombinant DNA!...

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BIMM100 Also Classic Expts: Chapter 5 -- Recombinant DNA Tues. (165-176); 176-187; Thurs. 188-198 Classic Experiments Finish: – repair & viruses (Genetics: terms & concepts) – the big (little) world of cloning Lecture 5
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Chapter 4 Finished - nucleic acids transcription translation Continuing - replication damage and repair viruses Chapter 5 Molecular Genetic Techniques (review 165-176) NOT COVERING HOMOLOGOUS RECOMBINATION(pp 150-153) http://www.hhmi.org/news/odonnell20100129.html Michael O'Donnell Rockefeller University “Whither the Replisome: Explanation for Why the Replisome Cannot Be Captured Alive” Wednesday, April 14th, 2010 4:00 pm/ Refreshments 3:45 pm CMG Large Conference Room The replisome is the kind of machine that any cartoonist has to love - flying apart and having to be constantly reassembled while moving and working at high speed. This is what Michael O'Donnell writes about his topic: "Replisomes were originally thought to be a multi-protein machine with a stabile and defined structure that is maintained during replication. Discovery that replisomes repeatedly discard sliding clamps and assemble a new clamp to start each Okazaki fragment provided the first hint that the replisome is more dynamic than originally envisioned. Recent studies reveal that the replisome is more dynamic than ever thought possible . .....
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Lecture 5 – Part I. Key Molecular Processes DNA Replication The process The players Coordination and bi-directional replication Finishing DNA Mistakes, Damage and Repair Polymerase – errors and proofreading A segue: Why we care A gallery of damage and repair deamination base excision mismatch repair UV damage and nucleotide excision repair global vs. transcription coupled NHEJ – last chance Viruses, briefly Host range Lytic vs. non-lytic life cycles The special case of retroviruses and Reverse Transcriptase Lecture 5 – Part II. Recombinant DNA Overview and Genetic concepts Cloning tools building a clone Libraries a source of clones screening Defining what you’ve cloned
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Eukaryotic Nucleotide Excision Repair: Global Genome Repair’ XPC and hHR23B dimer binds to damaged distorted DNA. TFIIH (XPB and XPD are helicases) binds to the complex. RPA and XPG bind, and the DNA is further unwound. The nucleases XPG and XPF/ERCC1 cleave the DNA. The oligonucleotide containing the damage is removed. DNA polymerase δ /( ε ) fill in the gap and ligase seals it.
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Eukaryotic Nucleotide Excision Repair: Transcription Coupled Repair (TCR) RNA polymerase stalls at the damaged site. CSB helps displace RNA Pol and recruit TFIIH (XPB and XPD), XPA, and RPA; the DNA is further unwound.
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mol5 - BIMM100! Lecture 5 ! Chapter 5 - Recombinant DNA!...

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