Difference between revisions of "Part:BBa K1017101"

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<partinfo>BBa_K1017101 short</partinfo>
 
<partinfo>BBa_K1017101 short</partinfo>
  
The chimera Cph8 is consist of the phytochrome Cph1 and the histidine kinase domain and response-regulator from EnvZ-OmpR. Cph8 autophosphorlates ompR, then phosphorylated-ompR activates the ompC promoter and turn on the downstream gene. Red light inhibits the autophosphorylation of Cph8 and turns off the gene expression. LacZ is the reporter gene.
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In general, the red light sensing promoter, OmpC promoter, turns on in the dark and turns off under red light with the present of Cph8.
 
[[File:Nctu_formosa_cph8mechnism.jpg|300px|center|link=]]
 
[[File:Nctu_formosa_cph8mechnism.jpg|300px|center|link=]]
 
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But we need to design a biobrick that can be turned on when exposing to red light. So we add LacI and lac promoter to downstream genes. So we design this biobrick that can detect red light, called red promoter.
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But we want to construct a more intuitive light regulating th system at can turn on under red light and turn off in the dark. So we created a new biobrick named red promoter by adding a lac operon to inverse the function of OmpC promoter.
  
 
Pred, including PompC,RBS(B0032), LacI, ter(J61048), and Plac, can be activated when exposed to red light.  OmpR, the transcription factor of PompC, is active when being phosphorylated in dark and inactive under red light. In the presence of red light, the non phosphorylated OmpR cannot turn on PompC. Thus, Plac won't be repressed by the negative regulator, LacI. So the whole Pred can express genes downstream Plac. On the other hand, if there is no red light, the phosphorylated OmpR then turn on PompC, causing LacI to be expressed and repress Plac to translate the genes downstream.
 
Pred, including PompC,RBS(B0032), LacI, ter(J61048), and Plac, can be activated when exposed to red light.  OmpR, the transcription factor of PompC, is active when being phosphorylated in dark and inactive under red light. In the presence of red light, the non phosphorylated OmpR cannot turn on PompC. Thus, Plac won't be repressed by the negative regulator, LacI. So the whole Pred can express genes downstream Plac. On the other hand, if there is no red light, the phosphorylated OmpR then turn on PompC, causing LacI to be expressed and repress Plac to translate the genes downstream.

Revision as of 13:34, 26 September 2013

Pred ( PompC + B0032 + LacI + J61048 + Plac)

In general, the red light sensing promoter, OmpC promoter, turns on in the dark and turns off under red light with the present of Cph8.

Nctu formosa cph8mechnism.jpg



But we want to construct a more intuitive light regulating th system at can turn on under red light and turn off in the dark. So we created a new biobrick named red promoter by adding a lac operon to inverse the function of OmpC promoter.

Pred, including PompC,RBS(B0032), LacI, ter(J61048), and Plac, can be activated when exposed to red light. OmpR, the transcription factor of PompC, is active when being phosphorylated in dark and inactive under red light. In the presence of red light, the non phosphorylated OmpR cannot turn on PompC. Thus, Plac won't be repressed by the negative regulator, LacI. So the whole Pred can express genes downstream Plac. On the other hand, if there is no red light, the phosphorylated OmpR then turn on PompC, causing LacI to be expressed and repress Plac to translate the genes downstream.

Nctu formosa pred.png


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 1282
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1296
  • 1000
    COMPATIBLE WITH RFC[1000]