1. Biotechnology l Introduction l Tools l Process l Applications

2. Biotechnology l Introduction – basic idea –Gene is identified and excised from one organism –Gene is placed in vector (plasmid) and amplified –Gene is transferred to new organism or used in host organism to make protein product

3. Biotechnology - Tools l Restriction endonucleases –Nucleases cut nucleic acid – at first seem non specific –Linn & Abner discover that some strains of bacteria are able to resist bacteriophage infection by digesting infecting DNA –Different bacteria produce different restriction enzymes

4. Biotechnology - Tools l Restriction Endonucleases –Cut at specific 4, 6, or 8 base sites –Site is a “palindrome” »Racecar »Madam I’m Adam »Damit I’m Mad –Some restriction enzymes generate “sticky ends”

5. Biotechnology - Tools

7. l Plasmids –Carry an antibiotic resistance marker –Carry restriction sites in convenient locations to insert DNA –Carry characteristics that allow the plasmid to reproduce in several organisms

9. Biotechnology - Tools

10. l Polymerase Chain Reaction (PCR) –Allows any segment of DNA to be amplified chemically in minutes –Uses a thermostable DNA polymerase –Machine can cycle every 60-90 seconds

11. Biotechnology - Tools

13. l Agarose Gel Electrophoresis –Can separate DNA fragments made with restriction enzymes –Can separate PCR made DNA –Can be used to sequence DNA

14. Biotechnology - Tools

15. Biotechnology - Process l Basic process worked out by Cohen & Boyer l Cut plasmid DNA and target DNA with same restriction enzyme l Mix DNA and allow sticky ends to match up l Select DNA clones having target gene

17. Biotechnology - Tools