Difference between revisions of "Part:BBa K3733005:Experience"
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===Applications of BBa_K3733005=== | ===Applications of BBa_K3733005=== | ||
| + | <p>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<sup>[1]</sup>. 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 <i>E. coli</i> DH5α.</p> | ||
| + | <p>In the experiment, we expand the culture of <i>E.coli</i> strains to OD<sub>600</sub>=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. </p> | ||
| + | <div><img></img></div> | ||
| + | <p><b>Figure 2. </b> OD<sub>600</sub> corrected RFU values culturing after 12 h after induction of different concentrations of sodium thiosulfate</p> | ||
===User Reviews=== | ===User Reviews=== | ||
Revision as of 00:54, 15 October 2021
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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
User Reviews
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