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Restriction Endonucleases David Peterson Structure of DNA and RNA Information Flow in Cells DNA RNA Transcription Protein Translation Amino acids combine to make peptides and proteins Enzymes • Catalyze reactions • Most are proteins Restriction endonucleases cut DNA at specific sites X174 DNA HaeIII HinfI Restriction Enzymes Aid in DNA Cloning Palindromes • Madam, I’m Adam. • Gary knits a stinky rag. • Harass sensuousness, Sarah. • Go hang a salami, I’m a lasagna hog. Most Restriction Enzymes Recognize Palindromes -GATATC-3' 3'-CTATAG-5' 5' EcoRV Bound to DNA www.pdb.org 1AZ0 Phage l 48,512 bp dsDNA virus that grows on E. coli. Life cycle ~ 30 min at 37°C typical burst ~ 100-300 particles Bacteriophage infection: 1 → 300 → 10,000 → 3,000,000 → 109! 30’ 30’ 30’ 30’ Phage DNA Bacterium 30 min Typical phage infection cycle: 1. Phage DNA penetrates bacterial envelope 2. Bacterial DNA is disrupted & phage DNA is replicated 3. Phage structural proteins are synthesized 4. Heads, tails, fibers made and assembled into particles 5. Cell lysis, release of ~ 100 – 300 phage particles Bacteriophages Make Plaques Werner Arber Dan Nathans and Hamilton Smith Nobel Prize in Physiology or Medicine 1978
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