Measurement

Part:BBa_K2447000

Designed by: Chee Wai Kit (david)   Group: iGEM17_NUS_Singapore   (2017-10-03)
Revision as of 00:29, 12 October 2017 by David135 (Talk | contribs)


Extracellular phosphate sensor with GFP reporter

PhoB and PhoR proteins are part of the Pho regulon inherent in E.coli. When low concentration of extracellular phosphate ions is present, PhoR can phosphorylate PhoB to form active phosphorylated-PhoB. PhoB promoter is activated as a result of active binding of phosphorylated-PhoB, resulting in downstream expression of GFP. When high concentration of extracellular phosphate ions is present, PhoR will dephosphorylate phosphorylated-PhoB, and therefore inactivating it, and repressing PhoB promoter for downstream expression of GFP.
PhoR and PhoB proteins work in tandem to control promoter PhoB and consequential downstream expression of GFP

Improvement over previous iGEM part BBa_K116404 (NYMUTaipei 2008)

The fluorescence of purified GFPmut3B was calibrated in the cell-free chassis. The derived [http://2007.igem.org/Imperial/Wet_Lab/Results/Res1.3 calilbration curve]allows the determination of the concentration of GFPmut3b in the cell-free chassis. [http://2007.igem.org/Imperial/Wet_Lab/Protocols/Prot1.3 Detailed protocols]for generating the calibration curve are available. Other calibration curves for are also available on the results page.
PhoR and PhoB proteins work in tandem to control promoter PhoB and consequential downstream expression of GFP

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PhoR and PhoB proteins work in tandem to control promoter PhoB and consequential downstream expression of GFP



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 BsaI.rc site found at 1162


[edit]
Categories
//function/reporter/fluorescence
Parameters
chassisE.coli (K12)