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Lab Meeting

Lab Meeting. 09/27/13. Program Update. Unafold arrived Need to install g77 complier. Theophylline Detection Results. How much Theophylline do cells need to grow on plates? . Test growth at 0.4, 0.2, 0.1, 0.05, 0.0 mM of Theophylline Look for visible GFP expression. psB1A2. High copy .

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Lab Meeting

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  1. Lab Meeting 09/27/13

  2. Program Update • Unafold arrived • Need to install g77 complier

  3. Theophylline Detection Results

  4. How much Theophylline do cells need to grow on plates? • Test growth at 0.4, 0.2, 0.1, 0.05, 0.0 mM of Theophylline • Look for visible GFP expression

  5. psB1A2 High copy Grow o/n @ 37C LB + Amp

  6. Colony streak culture onto plates LB + Amp 0.1mM Theo 100ug/ml Amp 0.05mM Theo 100ug/ml Amp 0.0mM Theo 100ug/ml Amp 0.4mM Theo 100ug/ml Amp 0.2mM Theo 100ug/ml Amp Grow o/n @ 37C

  7. 0.0mM Theophylline 0.05mM Theophylline 0.1mM Theophylline 0.2mM Theophylline 0.4mM Theophylline

  8. How much Theophylline do cells need to grow with a fixed amount of tetA? • Test growth at 0.4, 0.2, 0.1, 0.05, and 0.0 mM of Theophylline • Also, test growth with 10ug/mL of tetA +/- Amp • Looking for visible growth

  9. psB1A2 High copy Grow o/n @ 37C LB + AMP + Theophylline

  10. Colony streak culture onto plates LB + Amp + Theophylline 0.0mM Theo 100ug/ml Amp 10mg/ml tetA 0.1mM Theo 100ug/ml Amp 10mg/ml tetA 0.05mM Theo 100ug/ml Amp 10mg/ml tetA 0.4mM Theo 100ug/ml Amp 10mg/ml tetA 0.2mM Theo 100ug/ml Amp 10mg/ml tetA

  11. Colony streak culture onto plates LB + Amp + Theophylline 0.0mM Theo 10mg/ml tetA 0.1mM Theo 10mg/ml tetA 0.05mM Theo 10mg/ml tetA 0.4mM Theo 10mg/ml tetA 0.2mM Theo 10mg/ml tetA

  12. LB+TetA 0.0mM Theophylline 0.05mM Theophylline 0.1mM Theophylline 0.2mM Theophylline 0.4mM Theophylline

  13. LB+AMP+TetA 0.0mM Theophylline 0.05mM Theophylline 0.1mM Theophylline 0.2mM Theophylline 0.4mM Theophylline

  14. Conclusions • Cell expression of the reporter gene is robust in the presence of the theophylline • Therefore, ecdM8 is likely not being translated • Gst9 cloning a potential solution

  15. Future Work • Finish Program • Develop 10 minute Preview Presentation • Start Evolution Experiment

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