Difference between revisions of "Part:BBa K3989012"
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<img src="https://static.igem.org/mediawiki/parts/3/37/21_UZurich_EsaR_activation_repression.jpeg"> | <img src="https://static.igem.org/mediawiki/parts/3/37/21_UZurich_EsaR_activation_repression.jpeg"> | ||
− | <figcaption><b>Figure 1</b>. EsaR acts as either an activator or a repressor.</figcaption> | + | <figcaption><b>Figure 1</b>. EsaR acts as either an activator or a repressor.[1]</figcaption> |
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<img src="https://static.igem.org/mediawiki/parts/0/0f/21_UZurich_AHL_sensitivity.jpeg"> | <img src="https://static.igem.org/mediawiki/parts/0/0f/21_UZurich_AHL_sensitivity.jpeg"> | ||
− | <figcaption><b>Figure 2.</b> AHL-sensitivity of EsaR and its variant of promoter P<sub>esa</sub>S.</figcaption> | + | <figcaption><b>Figure 2.</b> AHL-sensitivity of EsaR and its variant of promoter P<sub>esa</sub>S. Different AHL concentrations are used: 0, 1nM, 10nM, 100nM, 1000nM and 10000nM.[1]</figcaption> |
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In this figure, we mainly show the distribution of the bacterium cells with a different fluorescence intensity. And it shows that except for I70V, other variants and wild-type have the same performance compared to our plate reader analysis. In I70V sample, the cell population is concentrated at a very low fluorescence intensity level and only few of the cells show a high intensity. After increasing the AHL concentration to 100nM, the population that have a high fluorescence intensity shift back to normal and when the concentration raises up to 1000nM, we can barely see any cells with high intensity. This performance might be caused by the quality of our sample because even there is no AHL molecule, there are only a few of cells show a high fluorescence intensity. Further reproduce experiment need to be performed. | In this figure, we mainly show the distribution of the bacterium cells with a different fluorescence intensity. And it shows that except for I70V, other variants and wild-type have the same performance compared to our plate reader analysis. In I70V sample, the cell population is concentrated at a very low fluorescence intensity level and only few of the cells show a high intensity. After increasing the AHL concentration to 100nM, the population that have a high fluorescence intensity shift back to normal and when the concentration raises up to 1000nM, we can barely see any cells with high intensity. This performance might be caused by the quality of our sample because even there is no AHL molecule, there are only a few of cells show a high fluorescence intensity. Further reproduce experiment need to be performed. | ||
+ | </html> | ||
+ | ===Sequence and Features=== | ||
+ | <partinfo>BBa_K3989012 SequenceAndFeatures</partinfo> | ||
+ | |||
+ | ===References=== | ||
+ | <html> | ||
+ | 1) Shong, J., Huang, Y. M., Bystroff, C., & Collins, C. H. (2013). Directed evolution of the quorum-sensing regulator EsaR for increased signal sensitivity. ACS chemical biology, 8(4), 789-795. | ||
</html> | </html> |
Latest revision as of 10:07, 17 October 2021
EsaR regulator
EsaR is a regulator protein which regulates the promoter of a bacterial Quorum Sensing system. By interacting with the Quorum Sensing molecules(a class of molecules called AHL.In our project, the molecule is called 3OC6HSL since it shows a higher sensitivity), it can either act as a repressor or an activator depending on the location of the promoter it binds.
How it works
In the Pesa promoter controlled Quorum Sensing system, EsaR acts as a regulator that can bind to a specific site of the promoter Pesa. Depending on the different types of the Pesa(PesaR and PesaS), EsaR can be either a repressor(for PesaR) or an activator(for PesaR). The detailed mechanism is shown below:
Characterisation
Previous studies has already characterised the 3OC6HSL sensitivity of the EsaR protein and its variants using a promoter activation assay. The result is shown below:
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
1) Shong, J., Huang, Y. M., Bystroff, C., & Collins, C. H. (2013). Directed evolution of the quorum-sensing regulator EsaR for increased signal sensitivity. ACS chemical biology, 8(4), 789-795.