Reporter

Part:BBa_I13556:Experience

Designed by: Austin Che   Group: iGEM2006_MIT   (2006-09-15)


Leakiness

It has been noticed that if this part is grown up in LB overnight in the absence of any inducer, the cells are strongly induced (up to about half the maximum fluorescence if AHL is added). This effect is not seen when the cells are grown up in EZ rich media or when the cells are made to grow slower. For example, there is a dramatic difference in the apparent leakiness of part depending on whether cells grown in LB are incubated at 37C (leaky) or 30C (not leaky). The root cause of this effect is unknown.

Transfer curve

Protocol

  1. Two colonies of pSB4K5.I13556 were grown up in 5ml LB Kan overnight
  2. Cultures were diluted 1:1000 and grown for 2 hours in 4ml EZ Rich medium + Kan
  3. 3 replicates of 100μl aliquots each for 8 AHL concentrations were made
  4. added 100μl EZ+AHL to 8 final concentrations of AHL: 0, 1E-11, 1E-10, 1E-9, 1E-8, 1E-7, 1E-6, 1E-5 M
  5. The cells were measured on a Victor3 fluorescence plate reader and grown overnight in the plate reader at 37C

Data

All data:

I13556-fluorescence.png

I13556-OD.png

Taking the last time point for all samples, we get the following transfer curve (note that the first time point labeled as 1E-12 is actually 0 AHL and was just set there for ease of plotting):

I13556-transfercurve.png

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