Part:BBa_K353002:Experience

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Applications of BBa_K353002

Part BBa_K353002 is modification of earlier parts characterizing the pBAD (BBa_I0500) promoter. It consists of pBAD (BBa_I0500) promoter, an RNA tag/RSID (K353010), GFP (E0040), and a terminator (B1006). The parts were characterized on plasmid 1C3 in BW27783 cells. GFP fluorescence data was collected on a plate reader overnight (99 reads every thirteen minutes). See dose-response curve for details. To gather flow cytometry data, fluorescence was read and plotted Below is a graph of our flow cytometry data for part K353002 plotting GFP output for BW27783 cells:

Both of these graphs plot the output of part K353002 in BW27783 cells induced with a series of concentrations of L-arabinose, measured in molarity. The first is a histogram showing the distribution of expression levels at each concentration of L-arabinose. The second is the same data displayed as a cumulative distribution so that population medians are more apparent.

We measured a dose-response-curve of median output vs L-arabinose concentration in triplicate. This curve constitutes the basic characterization of part K353002, including measurement of the concentration of L-arabinose which gives half-maximal output, the steepness of transition from off to on, and the parts dynamic range. Fitting a Hill curve provided the following values plus or minus 95% confidence intervals:

Half-max induction: 143 +- 36 uM

Hill coefficient: 1.4 +- .6

Fold Induction: 860

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