Part:BBa_K861169:Experience

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Device Design

In this design, we used promoter PcstA and an intermediate regulator cI, as PcstA is activated by CRP in low glucose concentration, whereas we need a device boosted by high concentration of glucose. As you can see, in this pathway, protein cI will be expressed when CRP is activated under low glucose concentration.The large amount of cI repressed the promoter R0051 heavily. On the contrary, when glucose concentration is high, the promoter is derepressed.

We made RFP as reporter and certify that the design works well via two different methods.

Fluorescence measurement

In the first method, Fluorescence intensity of cell culture was directly read from a plate reader. And all data is normalized by OD600.

In this figure, the fluorescence of RFP generator promoted by PcstA decreased when concentration of glucose rose, meanwhile, the fluorescence of the indirect regulatory device increased with the glucose concentration, indicating that the device works as expected.

The second method is practiced for single cells. The fluorescence pictures of different glucose concentration were captured by fluorescence microscope. Also, a program named FANCY was designed to recognize single cell and calculate the fluorescence strength in the images. As the result shows, it conformed well to that of the first method.

All these results indicated that the device we designed can meet the demand, promoters activated by glucose.

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