Difference between revisions of "Part:BBa K1886001"
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1886001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1886001 SequenceAndFeatures</partinfo>
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<span style="font-size:25px; font-weight:bold; border-bottom:2px solid #584b4f;">BACKGROUND</span></br></br>
 
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<span style="font-size:22px; font-weight:bold;">Overview</span>
 
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The synchronized oscillator design is based on elements of the quorum sensing machineries in Vibrio fischeri and Bacillus Thurigensis. We placed the luxI (from V. fischeri), aiiA (from B. Thurigensis) and yemGFP genes(which is used as a reporter and knocked out after the combination with logical gate) under the control of three identical copies of the luxI promoter. The LuxI synthase enzymatically produces an acyl-homoserine lactone (AHL), which is a small molecule that can diffuse across the cell membrane and mediates intercellular coupling. It binds intracellularly to the constitutively produced LuxR, and the LuxR–AHL complex is a transcriptional activator for the luxI promoter. AiiA negatively regulates the promoter by catalysing the degradation of AHL. This network architecture, whereby an activator activates its own protease or repressor, is similar to the motif used in other synthetic oscillator designs and forms the core regulatory module for many circadian clock networks. Furthermore, theoretical work has shown how the introduction of an autoinducer in similar designs can potentially lead to synchronized oscillations over a population of cells.
 
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<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K1886001 parameters</partinfo>
 
<partinfo>BBa_K1886001 parameters</partinfo>
 
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<h2>'''Characterization'''</h2>
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<h3> BACKGROUND </h3>
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<h4>Principle</h4>
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The synchronized oscillator design is based on elements of the quorum sensing machineries in Vibrio fischeri and Bacillus Thurigensis. We placed the luxI (from V. fischeri), aiiA (from B. Thurigensis) and yemGFP genes(which is used as a reporter and knocked out after the combination with logical gate) under the control of three identical copies of the luxI promoter. The LuxI synthase enzymatically produces an acyl-homoserine lactone (AHL), which is a small molecule that can diffuse across the cell membrane and mediates intercellular coupling. It binds intracellularly to the constitutively produced LuxR, and the LuxR–AHL complex is a transcriptional activator for the luxI promoter. AiiA negatively regulates the promoter by catalysing the degradation of AHL. This network architecture, whereby an activator activates its own protease or repressor, is similar to the motif used in other synthetic oscillator designs and forms the core regulatory module for many circadian clock networks. Furthermore, theoretical work has shown how the introduction of an autoinducer in similar designs can potentially lead to synchronized oscillations over a population of cells.
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<br>
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<h3>RESULTS</h3>
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<br>
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<h4>Gel electrophoretic analysis</h4>
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<br>
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[[File:RIRaiiA1016_副本.jpg|800px|thumb|center|Fig.1 Gel electrophoretic analyses of PCR products ]]
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<br>
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<br>
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<h4>stable oscillation in microfluidics chips</h4>
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[[File:Oscillationa.png|800px|thumb|left|Fig.2  ]]
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<br>
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<br>
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<br>

Latest revision as of 02:55, 20 October 2016


AHL single-cycled oscillation

Two molecules of luxR binds with AHL (catalyzed by luxI), and this compound will promote the activity of pluxR. pluxR controls the expression of gene aiiA , encoding the enzyme which degrades AHLs. And all of these form an AHL single-cycled oscillation.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1821
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 3256
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1101
    Illegal BsaI.rc site found at 1970
    Illegal BsaI.rc site found at 3073

Characterization

BACKGROUND

Principle

The synchronized oscillator design is based on elements of the quorum sensing machineries in Vibrio fischeri and Bacillus Thurigensis. We placed the luxI (from V. fischeri), aiiA (from B. Thurigensis) and yemGFP genes(which is used as a reporter and knocked out after the combination with logical gate) under the control of three identical copies of the luxI promoter. The LuxI synthase enzymatically produces an acyl-homoserine lactone (AHL), which is a small molecule that can diffuse across the cell membrane and mediates intercellular coupling. It binds intracellularly to the constitutively produced LuxR, and the LuxR–AHL complex is a transcriptional activator for the luxI promoter. AiiA negatively regulates the promoter by catalysing the degradation of AHL. This network architecture, whereby an activator activates its own protease or repressor, is similar to the motif used in other synthetic oscillator designs and forms the core regulatory module for many circadian clock networks. Furthermore, theoretical work has shown how the introduction of an autoinducer in similar designs can potentially lead to synchronized oscillations over a population of cells.

RESULTS


Gel electrophoretic analysis


Fig.1 Gel electrophoretic analyses of PCR products



stable oscillation in microfluidics chips

Fig.2