1. Restriction Endonucleases
David Peterson

2. Structure of DNA and RNA

3. Information Flow in Cells
DNA RNA Protein
Transcription
Translation

4. Amino acids combine to make peptides and proteins

5. Catalyze reactions Most are proteins
Enzymes
Catalyze reactions
Most are proteins

6. Restriction endonucleases cut DNA at specific sites
X174 DNA
HaeIII
HinfI

7. Restriction Enzymes Aid in DNA Cloning

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

10. Most Restriction Enzymes Recognize Palindromes
5'-GATATC-3'
3'-CTATAG-5'

11. EcoRV Bound to DNA
1AZ0

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

13. Bacteriophage infection: 1 → 300 → 10,000 → 3,000,000 → 109
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

14. Bacteriophages Make Plaques

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