Difference between revisions of "Part:BBa K2116005"
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[[File:T--ETH Zurich--p22.png|thumb|500px|Design of AND gate. The spacing between the esaboxes is 8bp long. NRB;NorR binding site, Sigma54B; RNA polymerase binding site]]
 
[[File:T--ETH Zurich--p22.png|thumb|500px|Design of AND gate. The spacing between the esaboxes is 8bp long. NRB;NorR binding site, Sigma54B; RNA polymerase binding site]]
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<p>
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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]].
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This Promoter (from here on refered to as PnorV) is the native promoter controlling the nitric oxide reduction operon (norRVW) in <i>E. coli</i> [1]. Its transcriptional regulator,
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NorR, can bind to nitric oxide and activate gene expression.
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Using the distinct properties of esaboxes, PnorV was designed to also be responsive to AHL, giving it an AND gate behaviour.
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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
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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
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were reached. We created and characterized a collection of these kind of AND gates: <br>
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<ul>
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<li> [[Part:BBa_K2116004]]
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<li> [[Part:BBa_K2116005]]
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<li> [[Part:BBa_K2116006]]
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<li> [[Part:BBa_K2116012]]
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<li> [[Part:BBa_K2116013]]
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<li> [[Part:BBa_K2116014]]
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<li> [[Part:BBa_K2116007]]
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<li> [[Part:BBa_K2116008]]
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<li> [[Part:BBa_K2116068]]
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<li> [[Part:BBa_K2116015]]
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</ul>
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</p>
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<h2> Characterization of this part </h2>
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Data will be here before the final presentation at the jamboree. Apologies for the delay.
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<h2>References:</h2>
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<ul>
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<li> [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>
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<li> [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. </li>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 14:34, 26 October 2016


AND gate regulated by norR and esaR (two esaboxes)

AND gate constructed by placing 2 esaboxes after the transcription start site of PnorV.

Design of AND gate. The spacing between the esaboxes is 8bp long. NRB;NorR binding site, Sigma54B; RNA polymerase binding site

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:


Characterization of this part

Data will be here before the final presentation at the jamboree. Apologies for the delay.

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]