Composite

Part:BBa_K381001

Designed by: Katharine Coyte   Group: iGEM10_BCCS-Bristol   (2010-10-05)

Nitrate reporter: PyeaR - GFP composite

Nitrate and Nitrite sensitive promoter PyeaR with a GFP coding device and strong RBS to create a nitrate-sensitive system which signals through expression of GFP.

PyeaR is normally repressed by NsrR, a protein native to most 'E. coli' cells. When Nitrate or Nitrite enter the cell it is converted to Nitric Oxide. This binds to NsrR, halting the repression and allowing the production of GFP.

Optimum use appears to be when sensing nitrate within a range of 0 - 20 mM (Calculated from Miller assays)

Activity between 0 - 10mM was investigated using an assay measuring change in GFP expression with increasing levels of Potassium Nitrate, the results of which can be seen below.

GFPChar0to10.png


ParisBettencourt12NitrateReporter.png

Additional characterization. Strain was NEB Turbo.


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 773


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