Difference between revisions of "Part:BBa K4948031:Experience"
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| − | + | =Usage and Biology= | |
| − | + | https://static.igem.wiki/teams/4948/wiki/parts/contribution/pa.jpg | |
| − | + | Figure 1. Optimized nsrR expression system and nitrate induced reporter system | |
| − | + | ||
| − | + | Our team's nitrate sensor (""BBa_K4948031"") was more sophisticated, based on the PyeaR promoter (""BBa_K216005"") from Edinburgh iGEM 2009. Since nsrR is a nitrate-dependent transcriptional repressor, a high amount of nsrR requires a high concentration of nitrate, resulting in a low performance sensor. Therefore, it is essential to determine the optimal concentration of nsrR. To this end, we used different constitutive expression units to control the intracellular concentration of nsrR. In addition, we added the hmpA1 site, which is known as the binding site of nsrR, between the yeaR promoter and the ribosome binding site. | |
| − | + | ||
| − | + | https://static.igem.wiki/teams/4948/wiki/parts/contribution/promoter-mscarlet.jpg | |
| − | + | Figure 2. Measuring the strength of a promoter with the reporter system | |
| − | + | ||
| + | |||
| + | https://static.igem.wiki/teams/4948/wiki/parts/contribution/f3.jpg | ||
| + | Figure 3. Changes in fluorescence intensity depending on the strength of the ribosome binding site in nitrate sensing | ||
| + | |||
| + | Our sensor is capable of detecting trace amounts of nitrate than existing sensors (Figure 4). To increase the intensity of nitrate-induced fluorescence, we added a strong RBS between the yeaR promoter and reporter gene, and in addition to this, we used a medium with higher nutrients to compensate for nitrate-induced cytotoxicity. | ||
| + | https://static.igem.wiki/teams/4948/wiki/parts/contribution/4.jpg | ||
| + | |||
| + | Figure 4. Fluorescence intensity by nitrate concentration | ||
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Latest revision as of 14:53, 12 October 2023
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_K4948031
User Reviews
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Usage and Biology
Figure 1. Optimized nsrR expression system and nitrate induced reporter system
Our team's nitrate sensor (""BBa_K4948031"") was more sophisticated, based on the PyeaR promoter (""BBa_K216005"") from Edinburgh iGEM 2009. Since nsrR is a nitrate-dependent transcriptional repressor, a high amount of nsrR requires a high concentration of nitrate, resulting in a low performance sensor. Therefore, it is essential to determine the optimal concentration of nsrR. To this end, we used different constitutive expression units to control the intracellular concentration of nsrR. In addition, we added the hmpA1 site, which is known as the binding site of nsrR, between the yeaR promoter and the ribosome binding site.
Figure 2. Measuring the strength of a promoter with the reporter system
Figure 3. Changes in fluorescence intensity depending on the strength of the ribosome binding site in nitrate sensing
Our sensor is capable of detecting trace amounts of nitrate than existing sensors (Figure 4). To increase the intensity of nitrate-induced fluorescence, we added a strong RBS between the yeaR promoter and reporter gene, and in addition to this, we used a medium with higher nutrients to compensate for nitrate-induced cytotoxicity.
Figure 4. Fluorescence intensity by nitrate concentration
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