1. Liquid cultures to Lysis: Movie Clip from Teachers domain Student Guide Lab 6

2. 1.What is made of DNA? 2.What is made of protein? 3.How is this related to rfp?

3. 1.What is made of DNA? 2.What is made of protein?

4. AraC dimer arabinose rfp Rfp protein rfp

5. RFP Bruce Wallace

6. RFP Bruce Wallace

7. Growth of transformed bacteria on various plates P+ plates P- plates LBLB/ampLB/amp/ara LBLB/amp No growth Bruce Wallace

8. The AraC Regulatory system

10. PreLab 6 Questions Lab 5a: Questions -Pass cultures in the room -Expectations -Why they grew this way? - All about the AraC operon

11. RFP expression araC generfp geneP BAD Transcription mRNA Translation araC protein Bruce Wallace

12. RFP expression rfp geneP BAD araC protein araC gene araC protein prevents RFP transcription by causing a loop to form in the region of the fp gene r Bruce Wallace NO ARABINOSE:

13. RFP expression araC protein arabinose araC generfp geneP BAD arabinose – araC protein complex RNA polymerase Arabinose – araC protein complex prevents DNA looping and helps to align RNA polymerase on the promoter site (P BAD ). mRNA Transcription Translation RFP (red fluorescent protein) Bruce Wallace With ARABINOSE:

14. Colony isolation and culture Preparing an overnight culture of E. Coli for RFP expression LB/amp/ara broth Bruce Wallace

15. Growth of transformed bacteria on various plates P+ plates P- plates LBLB/ampLB/amp/ara LBLB/amp No growth Bruce Wallace

17. Purification of RFP from an overnight culture Overnight culture Cell pellet with RFP Lysed cells Pellet cell debris RFP with binding buffer Bruce Wallace