Difference between revisions of "Part:BBa K3599019"

 
 
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<partinfo>BBa_K3599019 short</partinfo>
 
<partinfo>BBa_K3599019 short</partinfo>
  
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a modified green fluorescent protein, we used its sequence as a reporting gene to measure the intensity of the expression
 
  
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<!-- Add more about the biology of this part here
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The sensor device of Pa promoter ([[Part:BBa_K3599018|BBa_K3599018]]), using enhanced GFP ([[Part:BBa_K3599016|BBa_K3599016]]) as reporter.
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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===Sequence and Features===
 
<partinfo>BBa_K3599019 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3599019 SequenceAndFeatures</partinfo>
  
  
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===Design & Experience===
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===Functional Parameters===
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In iGEM2020 TPR_China Project, an aromatic sensor was constructed using Pa promoter, the eGFP ([[Part:BBa_K3599016|BBa_K3599016]]) and the sfGFP ([[Part:BBa_K3599017|BBa_K3599017]]) were both used as downstream reporters to form the sensor devices. To sense the aromatic compounds, PaaX under [[Part:BBa_K1031744|J23114]] constitutive promoter was used together.
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<partinfo>BBa_K3599019 parameters</partinfo>
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To characterize the original function of PaaX sensor device, we use 2-Hydroxyphenylacetic Acid (2HA) as the corresponding aromatic compond. According to the biological knowledge, PaaX could sense the intracellular concentrations of phenylacetyl-CoA, of which the chemical structure is semilar with 2HA.
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====The fluorescence of the sensor induced by small molecules====
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Through the corresponding small molecule sensing experiment, except Dmpr sensor, all our sensors have great induction effects on its corresponding aromatic small molecules. Among which NahR-sfGFP was the best! It is 63 times of the negative control.And here we can see that Xyls-sfGFP is 18 times of the Xyls-GFP( BBa_K1031911). And what surprises us is that Paax sensor also has a good response effect to small molecules, which has not been detected before.<br>
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[[File:T--TPR China--Sensor1.png|600px|thumb|center|The fluorescence of the sensor induced by small molecules]]<br>
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[[File:T--TPR China--Sensor2.png|600px|thumb|center|The fluorescence of the NahR sensor induced by small molecules]]<br>
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====The fluorescence of the sensor induced by PAN====
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And here is the fluorescence of different sensors induced by PAN.This data shows the same result that NahR-sfGFP responded best.
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[[File:T--TPR China--Sensor3.jpg|600px|thumb|center|The fluorescence of the sensor induced by PAN]]<br>

Latest revision as of 15:42, 27 October 2020


Pa-eGFP


The sensor device of Pa promoter (BBa_K3599018), using enhanced GFP (BBa_K3599016) as reporter.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 724
    Illegal BsaI.rc site found at 1581


Design & Experience

In iGEM2020 TPR_China Project, an aromatic sensor was constructed using Pa promoter, the eGFP (BBa_K3599016) and the sfGFP (BBa_K3599017) were both used as downstream reporters to form the sensor devices. To sense the aromatic compounds, PaaX under J23114 constitutive promoter was used together.

To characterize the original function of PaaX sensor device, we use 2-Hydroxyphenylacetic Acid (2HA) as the corresponding aromatic compond. According to the biological knowledge, PaaX could sense the intracellular concentrations of phenylacetyl-CoA, of which the chemical structure is semilar with 2HA.

The fluorescence of the sensor induced by small molecules

Through the corresponding small molecule sensing experiment, except Dmpr sensor, all our sensors have great induction effects on its corresponding aromatic small molecules. Among which NahR-sfGFP was the best! It is 63 times of the negative control.And here we can see that Xyls-sfGFP is 18 times of the Xyls-GFP( BBa_K1031911). And what surprises us is that Paax sensor also has a good response effect to small molecules, which has not been detected before.

The fluorescence of the sensor induced by small molecules

The fluorescence of the NahR sensor induced by small molecules

The fluorescence of the sensor induced by PAN

And here is the fluorescence of different sensors induced by PAN.This data shows the same result that NahR-sfGFP responded best.

The fluorescence of the sensor induced by PAN