1. Restriction Endonucleases David Peterson

2. Structure of DNA and RNA

3. DNA RNA Protein Information Flow in Cells

4. DNA is a Double Helix

6. Phage 48,512 bp dsDNA virus that grows on E. coli. Life cycle ~ 30 min at 37°C typical burst ~ 100-300 particles

7. Bacteriophage infection: 1 → 300 → 10,000 → 3,000,000 → 10 9 ! 30’ 30’ 30’ 30’ Bacterium Phage DNA 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 30 min

8. 1000’s of phage particles attacking an E. coli cell

9. Bacteriophages Make Plaques

11. Restriction Endonucleases Type 1 Cuts at random position relative to recognition sequence (sometimes far away) Type II Cuts at recognition sequence Type III Cuts at fixed distance from recognition sequence

12. SV40 DNA Cut with Hin cII

13. Palindromes Madam, I’m Adam. Gary knits a stinky rag. Harass sensuousness, Sarah. Go hang a salami, I’m a lasagna hog.

14. Most Restriction Enzymes Recognize Palindromes GATATC ||||||||||||||||||| CTATAG 5’ 3’

15. EcoRV Bound to DNA www.pdb.org 1AZ0

17. Methylation sites

18. Most Restriction Enzymes Recognize Palindromes GATATC ||||||||||||||||||| CTATAG 5’ 3’

19. EcoRV Bound to DNA www.pdb.org 1AZ0

20. SV40 DNA Cut with Hin cII

21. Restriction Enzymes Aid in DNA Cloning

22. Dan Nathans and Hamilton Smith Werner Arber Nobel Prize in Physiology or Medicine 1978