IGEM:MIT/2005/DeLong Discussion

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  • Rhod. in E.Coli, grabbing free proton from interior of cell, through shuttle mech. in retinal, producing proton.
  • Source of protons can come from anything (fermentation, example).
  • Rates: Can do calculations by looking at graphs. dpH in 5 minutes -.05 of 1.5 mL Eppendorf tubes. appx. -.01 pH drop/min.
  • Maybe engineer the host that you put it in.
    • Membrane vesicles possibly
    • We should do this: look at ATP pool in the cell (flux) to see if the proton pumping with light actually affects the nucleotide...(something)?
    • Can we get sustainable proton flux over reasonable amt time? Reasonable production?
      • Just a flash? No worries.
    • The proof of principle is more important than how sustainable the cell is. That can come later.
  • E.coli gets reducing power (NADH) from foodstuffs, fermentation from LB, perspiring aerobically, biochemical mechanisms that produce protons itself.
  • Need retinal in the media in order to express something and make it work.
  • Real concern whether you can take those protons and do something with it.