Difference between revisions of "Part:BBa K3989008"
Line 29: Line 29:
  
 
<html>
 
<html>
−
<. ><b>Figure 3.</b>Gene expression controlled by AHL molecules, EsaR regulator and P<sub>esaR</sub> promoter. On the  
+
<___><b>Figure 3.</b>Gene expression controlled by AHL molecules, EsaR regulator and P<sub>esaR</sub> promoter. On the  
 
left side, the test is based on the P<sub>esaR</sub> promoter and on the right side, the test is based on the P<sub>esaRC</sub> promoter. If one use 10nM
 
left side, the test is based on the P<sub>esaR</sub> promoter and on the right side, the test is based on the P<sub>esaRC</sub> promoter. If one use 10nM
 
</html>
 
</html>

Revision as of 14:51, 18 October 2021


Engineered quorum sensing esaR promoter: PesaR-C

A promoter that can interact with EsaR protein and inhibit downstream gene transcription.

Basic information

This part represents a promoter(Pesa) variant with two esa box, which can be bound to EsaR protein(see figure 2). esa box is a 18bps period located at a specific position of the promoter. After binding to the box, the EsaR will repress the recruitment of the RNA polymerase and inhibit the downstream transcription. However, when there are AHL molecules(such as 3OC6HSL) existing in the environment, they will interact with the EsaR and release it from the promoter, thus the transcription can be started.(Figure 1) The transcription level is controlled by the concentration of the AHL molecules.

Figure 1. Mechanism of the EsaR being an repressor under the control of the original PesaR. The red dots are the specific AHL molecule and in our project it is 3OC6HSL[1]
Below is the figure of the structure of the original promoter and the engineered one.
Figure 2. The structure of PesaR-C and the original PesaR.[2]
As we can see in figure 2, the original promoter: PesaR has only one esa box located at position -10 and the PesaR-C has an additional esa box which locates between position -10 and -35. Previous studies has shown that even with a same amount of AHL molecules and same type of the EsaR regulator, the gene expression level can still be different depending on the type of the promoter.(see figure 3)


21 UZurich PeaR charac.jpeg21 UZurich PeaRC charac.jpeg

<___>Figure 3.Gene expression controlled by AHL molecules, EsaR regulator and PesaR promoter. On the left side, the test is based on the PesaR promoter and on the right side, the test is based on the PesaRC promoter. If one use 10nM Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 281
    Illegal XhoI site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]