Difference between revisions of "Part:BBa K2116004"
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<h2> Characterization of this part </h2>
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<h1> Biology and Usage </h1>
  
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<p> This AND gate has one esabox placed as a roadblock after PnorV. [[File:T--ETH Zurich--p21new.png|thumb|500px;]] </p>
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<p>Biological logic gates are useful for creating higher order genetic circuits. This AND gate has one esabox placed as a roadblock
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after PnorV transcription start site. [[File:T--ETH Zurich--p21new.png|thumb|500px;]] It is regulated by a transcriptional
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activator, NorR, and a transcriptional repressor, EsaR. Transcription can be initiated by NO binding to NorR. EsaR sits
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on the esabox and blocks RNA polymerase from advancing. As soon as 3OC6HSL binds EsaR it is released and transcription can
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continue. This design makes the AND gate modular. The esabox/EsaR system can be exchanged for another transcriptional repression
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system to create another AND gate.
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</p>
  
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Data will be here before the final presentation at the jamboree. Apologies for the delay.
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<h1>Characterization</h1>
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<p>When characterizing our parts collection we initially confirmed functionality. Below you can see a graph depicting AND gate behaviour of this biobrick. [[]]
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</p>
  
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<p>Explain the graph, maybe refer to mechanics.</p>
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<p>Secondly we tested the system with and without EsaR present, in order to show that the AND gate behaviour is due to repression by EsaR. [[]] Here we show that  </p>
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<p></p>
 
<h2>References:</h2>
 
<h2>References:</h2>
  

Revision as of 22:16, 27 October 2016

AND gate regulated by norR and esaR (one esabox)

We constructed a selection of AND gates responding to nitric oxide (NO) and 3OC6HSL (AHL). They were designed using the previously described NorV promoter Part:BBa_K1153000. This Promoter (from here on refered to as PnorV) is the native promoter controlling the nitric oxide reduction operon (norRVW) in E. coli [1]. Its transcriptional regulator, NorR, can bind to nitric oxide and activate gene expression. Using the distinct properties of esaboxes, PnorV was designed to also be responsive to AHL, giving it an AND gate behaviour. An esabox is an 18bp sequence to which the transcriptional regulator EsaR Part:BBa_K2116001 can bind. Transcription can be initiated by the specific AHL EsaR responds to [N-(3-oxo-hexanoyl)-L-homoserine lactone]. By placing one, two or three esaboxes at different positions in the vicinity of PnorV, different specificities for AHL and NO were reached. We created and characterized a collection of these kind of AND gates:


Biology and Usage

Biological logic gates are useful for creating higher order genetic circuits. This AND gate has one esabox placed as a roadblock after PnorV transcription start site.

500px;
It is regulated by a transcriptional

activator, NorR, and a transcriptional repressor, EsaR. Transcription can be initiated by NO binding to NorR. EsaR sits on the esabox and blocks RNA polymerase from advancing. As soon as 3OC6HSL binds EsaR it is released and transcription can continue. This design makes the AND gate modular. The esabox/EsaR system can be exchanged for another transcriptional repression system to create another AND gate.

Characterization

When characterizing our parts collection we initially confirmed functionality. Below you can see a graph depicting AND gate behaviour of this biobrick. [[]]


Explain the graph, maybe refer to mechanics.

Secondly we tested the system with and without EsaR present, in order to show that the AND gate behaviour is due to repression by EsaR. [[]] Here we show that


References:


  • [1] Gardner, A. M. "Regulation Of The Nitric Oxide Reduction Operon (Norrvw) In Escherichia Coli. ROLE OF Norr AND Sigma 54 IN THE NITRIC OXIDE STRESS RESPONSE". Journal of Biological Chemistry 278.12 (2003): 10081-10086.</i>
  • [2] Shong, Jasmine and Cynthia H. Collins. "Engineering The Esar Promoter For Tunable Quorum Sensing-Dependent Gene Expression". ACS Synth. Biol. 2.10 (2013): 568-575.


    • 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
      COMPATIBLE WITH RFC[1000]