Difference between revisions of "Part:BBa K2548001"

 
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<partinfo>BBa_K2548001 short</partinfo>
 
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<p>RhlR coding device with 60 times of predicted RBS strength compared with BBa_K2548000. The RBS sequence is obtained from Salis Lab RBS Calculator V2.0.</p>
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<h2> Characterization of Sensor Device III </h2>
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            <p>We calculated four new RBS sequence with strength of 10, 20, 30, and 60 times of BBa_B0034 and assembled them with our sensor device. We tested the GFP production rate of these four devices, and found the device with 60 times of RBS strength gave the best result. The GFP production rate of this device at 9.90E-8 M of C4-HSL is:</p>
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            https://static.igem.org/mediawiki/2018/4/4b/T--BNDS_CHINA--Demo11.png
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                https://static.igem.org/mediawiki/2018/1/1e/T--BNDS_CHINA--Demo6.png
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            <p>This time, the difference between GFP production rate at 0 M and 9.90E-8 M of C4-HSL reached 35.7% of the expression level without induction, enabling people to detect <i>A. hydrophila</i> more easily from fish feces by using fluorescence detector. Moreover, as Mr. Zhang predicted before, the cooperativity between RhlR and C4-HSL was improved. Figure x. shows the modelling results of the three sensor devices. The slope of device III is the steepest one, and the Hill coefficient of the function, which describes the cooperativity between inducer and RhlR, increased from 1.154 in device I to 2.250 in this device.</p>
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            https://static.igem.org/mediawiki/2018/b/be/T--BNDS_CHINA--Demo13.png
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RhlR coding device with 60 times of predicted RBS strength compared with BBa_K2548000. The RBS sequence is obtained from Salis Lab RBS Calculator V2.0.
 
  
 
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Revision as of 19:07, 17 October 2018


Constitutive promoter- RBS - rhlR (LVA tagged) -terminator

RhlR coding device with 60 times of predicted RBS strength compared with BBa_K2548000. The RBS sequence is obtained from Salis Lab RBS Calculator V2.0.

Characterization of Sensor Device III

We calculated four new RBS sequence with strength of 10, 20, 30, and 60 times of BBa_B0034 and assembled them with our sensor device. We tested the GFP production rate of these four devices, and found the device with 60 times of RBS strength gave the best result. The GFP production rate of this device at 9.90E-8 M of C4-HSL is:

           T--BNDS_CHINA--Demo11.png
               T--BNDS_CHINA--Demo6.png

This time, the difference between GFP production rate at 0 M and 9.90E-8 M of C4-HSL reached 35.7% of the expression level without induction, enabling people to detect A. hydrophila more easily from fish feces by using fluorescence detector. Moreover, as Mr. Zhang predicted before, the cooperativity between RhlR and C4-HSL was improved. Figure x. shows the modelling results of the three sensor devices. The slope of device III is the steepest one, and the Hill coefficient of the function, which describes the cooperativity between inducer and RhlR, increased from 1.154 in device I to 2.250 in this device.

           T--BNDS_CHINA--Demo13.png


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 BamHI site found at 295
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 770