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pGLO plasmid

pGLO plasmid. Beta Lactamase Ampicillin resistance Green Fluorescent Protein (GFP) Aequorea victoria jellyfish gene araC regulator protein Regulates GFP transcription. ara Operon. lac Operon. araC. B. A. D. LacI. Z. Y. A. Effector (Arabinose). Effector (Lactose). araC. B. A.

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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

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