Difference between revisions of "Part:BBa K3245002"
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<h2>Characterization:</h2>
 
<h2>Characterization:</h2>
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<p>To test this part’s function, we cultured DH10B bacteria carried this plasmid with another downstream luxpr-GFP plasmid at the same time. Luxpr is a promoter that can be activated in QS system. By measuring fluorescent/OD600, the expressing level of luxpr can be known. We cultured the bacteria above overnight. As it reached a high concentration, luxpr is keeping activated and highly expressing GFP. Then we diluted the culture and measured its fluorescent and OD 600 every 30 minutes. The first measurement shows luxpr hasn’t be shut immediately, keeping the fluorescent/OD600 at a high level. But as the concentration of signal is low in the fresh culture, luxpr is gradually shut in the next few hours and the fluorescent/OD keeps going down. After the newly cultured bacteria grow to a higher concentration, luxpr started to be activated and fluorescent/OD600 keeps up again as the following picture shows. This proves this part can regulate the express level of luxpr and works in QS system.</p>
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<p><a href="https://parts.igem.org/Part:BBa_R0062">For more information: Characterization of BBa_R0026</a></p>
  
  

Revision as of 09:23, 20 October 2019

BBa_K3245002:

Usage and biology:

In QS system, the upstream product gets higher concentration as the population grows, which then regulates activity of the downstream promotor. In this part, LuxR activator protein combines with homoserine lactone(HSL) produced by LuxI protein as dimers. The dimers then up-regulate the expression of luxpr.

Design:

QS system is the main regulating circuit in our project. After comparing the efficiency of different QS system，we chose LuxI and LuxR to regulate the downstream circuit. To balance and stabilize the copy number of luxI (BBa_C0061) and luxR(BBa_C0062), we designed this plasmid carrying the two gene at the same time. Both gene uses the J23106 promoter and B0015 terminator. As a upstreaming plasmid for inducing, its measurement needs to cooperate with another downstream QS promotor (luxpr-GFP in our experiment).


Characterization:

<a href="https://parts.igem.org/Part:BBa_R0062">For more information: Characterization of BBa_R0026</a>


T--Fudan--Part20.png

Fig,1 this figure shows the WT luxpr expression level induced by luxR and luxI. The MEFL/particle changes as the OD goes up after dilution.



Final dimeric production regulating luxpr

In QS system, the upstream product gets higher concentration as the population grows, which then regulates activity of the downstream promotor. In this part, LuxR activator protein combines with homoserine lactone(HSL) produced by LuxI protein as dimers. The dimers then up-regulate the expression of luxpr.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 678
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]