Regulatory

Part:BBa_K1082002

Designed by: Gunvor Røkke   Group: iGEM13_NTNU-Trondheim   (2013-09-23)

Pm/XylS promoter system

The Pm/XylS promoter system consists of a coding part for XylS and the Pm promoter. When induced with m-toluic acid, the m-toluic acid binds to the XylS protein in order to create an inducer complex that in turn activates the Pm promoter.

We tested the promoter according to this article. The absorbance was measured at 600 nm, excitation at 485 nm and emission at 525 nm. Background fluorescence was also determined by measuring only media and this measurement was substracted from the measurements of the samples.

The Pm/xylS is positively regulated by m-toluic acid, so the samples were added different concentrations (0nM, 100nM, 1µM, 10µM, 100µM, 1mM and 2mM) to see the effect. (figure X). The fluorescence and absorbance measurements were used to calculate the ratio of GFP synthesis. The results show that the GFP are expressed in the samples with the Pm/xylS promoter, indicating that the promoter works as it should.

The figure shows the ratio of GFP synthesis as a function of inducer concentration.

Pmxylsprom.png


Design Notes

Source

The wiki of NTNU_Trondheim 2013 team

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 930
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 208
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 745
    Illegal BsaI site found at 1220


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