Difference between revisions of "Part:BBa K1767001"
 
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<partinfo>BBa_K1767001 short</partinfo>
 
<partinfo>BBa_K1767001 short</partinfo>
  
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We designed and built a kill switch mechanism that is based on the activation of a lethal gene downstream of an inducible promoter that is activated over time. The presented part is the first module of our designed genetic circuit (BBa_K1767003). In order to characterize and understand each module of the genetic circuit we designed two composite parts that allow us to verify the (1) the degradation rate of AHL by AiiA and (2) induction of the pLux promoter by LuxR in presence of AHL with and without co-expression of AiiA.
 
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(1) The here presented BioBrick consists of a constitutive promoter driving expression of AiiA and LuxR followed by a Lux promoter which drives expression of a fluorescent reporter (here: yellow fluorescent protein). For a schematic overview see the figure below.
 
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The idea is to retrieve information about the expression of a fluorescent reporter downstream of the pLuxR promoter in absence and increasing concentrations of AHL. AiiA degrades the AHL molecules hence limiting the amount of AHL available to the cells and more specifically to activate LuxR.
 
  
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<p>We designed and built a kill switch mechanism that based on the activation of a lethal gene downstream of an inducible promoter that can be activated over time.</p>
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<p>The presented part is the first module of our designed genetic circuit. In order to characterize and understand each module of the genetic circuit we designed two composite parts that allow us to verify the </p>
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<p>(1) the degradation of AHL by AiiA and</p>
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<p>(2) induction of the pLux promoter by LuxR in presence of AHL.</p>
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<p>The here presented BioBrick consists of a constitutive promoter (I14032) driving expression of Aiia and LuxR followed by a Lux promoter (R0062). </p>
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<p>The idea is to retrieve information about the expression of a fluorescent reporter downstream of the pLuxR promoter in absence and increasing concentrations of AHL. Aiia degrades the AHL molecules so it limits the amount of AHL that we have in the bacteria and hence from the active LuxR protein.The scheme below explains how this BioBrick works :</p>
  
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[[File: Technion HS Israel K1767001.jpg|450px|center]]
  
  

Latest revision as of 10:26, 16 September 2015

P(Lac)IQ RBS Aiia RBS LuxR term term luxpR


We designed and built a kill switch mechanism that based on the activation of a lethal gene downstream of an inducible promoter that can be activated over time.

The presented part is the first module of our designed genetic circuit. In order to characterize and understand each module of the genetic circuit we designed two composite parts that allow us to verify the

(1) the degradation of AHL by AiiA and

(2) induction of the pLux promoter by LuxR in presence of AHL.

The here presented BioBrick consists of a constitutive promoter (I14032) driving expression of Aiia and LuxR followed by a Lux promoter (R0062).

The idea is to retrieve information about the expression of a fluorescent reporter downstream of the pLuxR promoter in absence and increasing concentrations of AHL. Aiia degrades the AHL molecules so it limits the amount of AHL that we have in the bacteria and hence from the active LuxR protein.The scheme below explains how this BioBrick works :


Technion HS Israel K1767001.jpg


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 157
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1761