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Experiments for the Output Amplification Model

Output Amplification Model

  1. Determine the maximum concentration of sD that
    cells can produce (in vivo): Compare activity with 2.
  2. Determine how much XylE we need to produce
    prior to adding Catechol (in vitro first, maybe in vivo
    afterwards). In vivo IPTG: Compare cultures that
    have less of it.
  3. Determine XylE and TEV production (in vivo).
    Use robot to induce production and measure
    activities TEV (FRET pairs with TEV link).
  4. Degradation of XylE (in vivo or in vitro if cell division
    is not taken into account). Monitoring activity, then
    approximate the concentration. Remove the IPTG.
  5. Determine TEV kinetics (kcat, Km) on
    XylE-GFP (in vitro).

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