1. Bioengineering Turning Genes on and off in Bacteria

3. If all cells have the same DNA then why aren’t all cells the same?

4. Gene Expression  Process of turning genes on and off in cells.  Allows for differentiation.  Gene expression is different in prokaryotes and eukaryotes:  Prokaryotes: operons  Eukaryotes: DNA modification, alternative splicing, transcription factors, RNAi

5. Question?  How can we get a bacteria to express a jellyfish gene under specific conditions ?

6. Problem #1:  How do we get a jellyfish gene into bacteria?  Design a plasmid:

7. Plasmid Design  Origin of replication:  Antibiotic resistance:  Promoter and restriction site within operon:

8. Heat Shock  Method to get bacteria to take in DNA. Bacteria take in DNA when they are in stressful environments!

9. How do bacteria express DNA that is not theirs?  Operon: Sequence of genes that it turned on by the presence of a specific chemical

10. We are going to clone a bacteria that is tricked into making green fluorescence protein!

11. pGLO lab

12. Introductory questions to consider  Where is the gene originally from?  What does the plasmid contain?  How will we get the plasmid into the bacteria?  What will activate the production of GFP in bacteria?  Google BioRAD and pGLO and find the manual and answer these questions prior to Lab next Wednesday!

13. Answers  Where is the gene originally from?  Green Fluorescence Protein from Jellyfish  What does the plasmid contain?  Ampicillin resistance, Arabinose operon  How will we get the plasmid into the bacteria?  Heat Shock  What will activate the production of GFP in bacteria?  Presence of arabinose

14. Hypothesis:  Visual: What will the four Petri dishes that we will make look like?  Written: What will cause the production of GFP in bacteria?