Difference between revisions of "Part:BBa K1031422"

 
 
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<partinfo>BBa_K1031422 short</partinfo>
 
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<p>For detailed information concerning HcaR and Ph promoter, please visit <a href="http://2013.igem.org/Team:Peking/Project/BioSensors/HcaR">2013 Peking iGEM Biosensor HcaR</a></p>
  
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Ph(HcaR)-B0034-sfGFP
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===Usage and Biology===
 
  
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== '''Structure''' ==
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<img src="https://static.igem.org/mediawiki/igem.org/4/41/Peking2013_HcaRFig1.jpg" style="width:600px; margin-left:170px" />
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<p style="text-align:center"><b>Fig 1</b> Structure of <i>Ph</i> promoter regulated by HcaR and chemical pathway.
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== '''Sequence and Features''' ==
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1031422 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1031422 SequenceAndFeatures</partinfo>
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== '''Construction and data''' ==
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For the biosensor circuit ''Ph''/HcaR, we created a library of RBS (Ribosome Binding Site) including B0031, B0032 and B0034 for tuning the expression level of reporter gene sfGFP.
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K1031422 is composed of three elements, ''Ph'' promoter activated by HcaR, RBS <html><a href="https://parts.igem.org/Part:BBa_B0034">B0034</a></html> and reporter gene sfGFP ('''Fig 1a'''). Induction ratio was tested when K1031422 is in collocation with J23106-HcaR, J23109-HcaR, J23113-HcaR and J23114-HcaR, as shown in the dashed boxes ('''Fig 1b'''). From data we concluded biosensor circuit ''Ph''/HcaR adopting ''Pc'' J23106 and RBS B0034 obtained the optimal performance.
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<img src="https://static.igem.org/mediawiki/2013/thumb/9/94/Peking2013_part_B0034_Ph_HcaR.png/800px-Peking2013_part_B0034_Ph_HcaR.png", 
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<p style="text-align:center"><b>Fig 1</b> Construction and tunning of reporter circuit <i>Ph</i>-B0034-sfGFP. (<b>a</b>) The orange arrow represents <i>Ph</i> promoter activated by HcaR. The green oval stands for RBS B0034. sfGFP coding sequence is shown with dark blue, while terminator B0015 is in dark red. (<b>b</b>) Test was done following <a href="http://2013.igem.org/Team:Peking/Team/Notebook/Protocols">Test Protocol 1</a>. Horizontal axis represents biosensor circuit adopting different combination of <i>Pc</i> promoter and RBS. Vertical axis stands for induction ratio. The dashed boxes refer to data for reporter circuit <i>Pc</i>-B0034-sfGFP in collocation with biosensor circuits J23106-HcaR and J23114-HcaR respectively.
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Latest revision as of 20:03, 28 October 2013

Ph(HcaR)-B0034-sfGFP

For detailed information concerning HcaR and Ph promoter, please visit 2013 Peking iGEM Biosensor HcaR


Structure

Fig 1 Structure of Ph promoter regulated by HcaR and chemical pathway.


Sequence and Features

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 SapI.rc site found at 113


Construction and data

For the biosensor circuit Ph/HcaR, we created a library of RBS (Ribosome Binding Site) including B0031, B0032 and B0034 for tuning the expression level of reporter gene sfGFP. K1031422 is composed of three elements, Ph promoter activated by HcaR, RBS B0034 and reporter gene sfGFP (Fig 1a). Induction ratio was tested when K1031422 is in collocation with J23106-HcaR, J23109-HcaR, J23113-HcaR and J23114-HcaR, as shown in the dashed boxes (Fig 1b). From data we concluded biosensor circuit Ph/HcaR adopting Pc J23106 and RBS B0034 obtained the optimal performance.

Fig 1 Construction and tunning of reporter circuit Ph-B0034-sfGFP. (a) The orange arrow represents Ph promoter activated by HcaR. The green oval stands for RBS B0034. sfGFP coding sequence is shown with dark blue, while terminator B0015 is in dark red. (b) Test was done following Test Protocol 1. Horizontal axis represents biosensor circuit adopting different combination of Pc promoter and RBS. Vertical axis stands for induction ratio. The dashed boxes refer to data for reporter circuit Pc-B0034-sfGFP in collocation with biosensor circuits J23106-HcaR and J23114-HcaR respectively.