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pGLO plasmid

pGLO plasmid

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pGLO plasmid

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  1. pGLO plasmid • Beta Lactamase • Ampicillin resistance • Green Fluorescent Protein (GFP) • Aequorea victoria jellyfish gene • araC regulator protein • Regulates GFP transcription

  2. ara Operon lac Operon araC B A D LacI Z Y A Effector (Arabinose) Effector(Lactose) araC B A D LacI Z Y A RNA Polymerase RNA Polymerase B A D araC Z Y A Need to understand Lactose and Arabinoseoperon

  3. ara GFP Operon ara Operon araC GFP Gene araC B A D Effector(Arabinose) Effector (Arabinose) araC B A D araC GFP Gene RNA Polymerase RNA Polymerase B A D araC araC GFP Gene Took Araoperon and put it into a plasmid/ replace BAD with GFP

  4. VERY IMPORTANT TO UNDERSTAND THIS SLIDE • Ampicillin (Amp) tests for the uptake of plasmid • Cells transformed with plasmid will GROW on Amp • Arabinose (Ara) removes repressor and express GFP • Cells transformed with plasmid and given Arabinose will GLOW on Amp + Ara

  5. What will happen on these plates?Experiment 5.2 • Answer with die, grow or glow • Bacteria that were not transformed • LB • LB + Amp • LB + Amp + Ara • Bacteria that were transformed with pGLO plasmid. • LB • LB + Amp • LB + Amp + Ara

  6. Experiment 7.3 – isolate GFP protein • What plates should we use? • LB only, LB + Amp or LB + Amp + Ara • What were these plates grown on?

  7. What will happen on these plates?Experiment 5.1 • Answer with die, grow or glow • Bacteria that were not transformed • LB • LB + Amp • Bacteria that were transformed with S3 plasmid. • LB • LB + Amp