Part:BBa_K3733005:Experience

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Applications of BBa_K3733005

It is known from the literature that the threshold for the two-component system to sense the corresponding signal molecule can be changed by changing the strength of the promoter and RBS^[1]. At the same time, the weaker the promoter and RBS used to express the proteins in the two-component system, the lower the threshold for sensing signal molecules. In order to enable our engineered bacteria to sense and respond to different concentrations of thiosulfate, we used J23106a as a promoter to control the expression of membrane protein ThsS(BBa_K2507000) and transcription factor ThsR(BBa_K2507001) in the two-component system ThsSR in E. coli DH5α.

In the experiment, we expand the culture of E.coli strains to OD₆₀₀=0.4, spot the bacteria liquid in a 96-well plate, and add a series of concentration gradient sodium thiosulfate solutions to induce fluorescence expression. RFU is measured by Synergy H1 microplate reader, and the gain is 50.

Figure 2.OD₆₀₀ corrected RFU values culturing after 12 h after induction of different concentrations of sodium thiosulfate

[1] Woo S G, Moon S J, Kim S K, et al. A designed whole-cell biosensor for live diagnosis of gut inflammation through nitrate sensing[J]. Biosensors and Bioelectronics, 2020, 168: 112523.

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