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Fluorescence is measured over time, and compared to fluorescene generated while using eYFP as a reporter  <a href="https://parts.igem.org/Part:BBa_K23870232"> (BBa_K2387032)</a>.
 
Fluorescence is measured over time, and compared to fluorescene generated while using eYFP as a reporter  <a href="https://parts.igem.org/Part:BBa_K23870232"> (BBa_K2387032)</a>.
  
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                                                     <b>Figure E:</b> CpxR dimerization visualized using eYFP (left) and mVenus (right), with L-arabinose concentration = 0.2% and different activator concentrations over time
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                                                     <b>Figure 1:</b> CpxR dimerization visualized using eYFP (left) and mVenus (right), with L-arabinose concentration = 0.2% and different activator concentrations over time
  
 
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More information can be found <a href="http://2017.igem.org/Team:Wageningen_UR/Results/SpecificVisualization">here</a>
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Latest revision as of 12:54, 1 November 2017


CpxR-mVenus[1-157] and CpxR-mVenus[158-238] + araC/pBAD promoter

The [http://parts.igem.com/Part:BBa_K2387038 N-terminus of mVenus] and the [http://parts.igem.com/Part:BBa_K2387039 C-terminus of mVenus] are fused to Cpx response regulator [http://parts.igem.com/Part:BBa_K2387002 CpxR] in order to visualize the activation of the Cpx pathway. Upon activation of the Cpx pathway, CpxR gets phosphorylated by E. coli endogenous CpxA after which it can homodimerize. This protein-protein interaction can be visualized using BiFC, hence the fusion of mVenus[1-157] and mVenus[158-238]. The fusion is put under the control of the L-arabinose inducible araC/pBAD promoter. Strong RBS BBa_B0034 is used to regulate transcription.

CpxR-mVenus[1-157] and CpxR-mVenus[158-238] fusions were linked together using a [http://parts.igem.com/Part:BBa_K1486004 Flexible Linker] consisting of two times the amino acids GGGGS.

This part is used to visualize the activation of the Cpx pathway via Bimolecular Fluorescence Complementation by using the CpxR-CpxR interaction.

Usage and Biology

This BioBrick was used following this protocol. Fluorescence is measured over time, and compared to fluorescene generated while using eYFP as a reporter (BBa_K2387032).

Figure 1: CpxR dimerization visualized using eYFP (left) and mVenus (right), with L-arabinose concentration = 0.2% and different activator concentrations over time

More information can be found here



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal XhoI site found at 1283
    Illegal XhoI site found at 2498
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1694
    Illegal AgeI site found at 2909
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961