Composite

Part:BBa_K353002:Experience

Designed by: Alejandro Virrueta   Group: iGEM10_Stanford   (2010-10-25)
Revision as of 01:04, 5 November 2010 by Anshurama (Talk | contribs) (→‎Applications of BBa_K353002)

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

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) after induction with varying does of arabinose. See dose-response curve for details. To gather flow cytometry data, fluorescence was read and plotted with number of cells as a function of intensity. The arabinose induction concentrations are provided as percentages. The top graph represents a fluorescence read for a single sample while the bottom graph represents a normalized fluorescence reading for the same data.

Below is a graph of our flow cytometry data for part K353002 plotting GFP output for BW27783 cells: Realdata.jpg

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.

740px-Flow data.jpg

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

User Reviews

UNIQe7320180f6b8f7ba-partinfo-00000000-QINU UNIQe7320180f6b8f7ba-partinfo-00000001-QINU