IGEM:Imperial/2010/Experiments1

 Experiments for the Output Amplification Model

Output Amplification Model

 Determine the maximum concentration of sD that cells can produce (in vivo): Compare activity with 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.  Determine XylE and TEV production (in vivo). Use robot to induce production and measure activities TEV (FRET pairs with TEV link).  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.  Determine TEV kinetics (kcat, Km) on XylE-GFP (in vitro). 