Reporter

Part:BBa_K2905016

Designed by: David Viguilla   Group: iGEM19_Alma   (2019-10-15)


High pH Sensor

This part is to be used as a reporter gene that is active at high pH levels.

Measuring Expression/Fluorescence at different pH: Alma 2019

To determine if the part works as intended (expression at high pH, repression at pH less than 5.5), K2905016 (this part) or a control, BBa_J04450 were analyzed at different pH. Cells were grown to mid-log in LB, then pelleted and transferred to M9 minimal media with pH adjusted to ~4.5 or left untouched at 7, and then grown for about 14 hours shaking at 37C. Fluorescence was measured in a Mx3500.

T--Alma--high pH graph1.png

As can be seen from the graph, there is a statistically significant difference between expression for the reporter when comparing acidic and neutral conditions. From this we would conclude that the reporter is working as intended, at least qualitatively - compare this data to the inverse part, BBa_K2762014.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1661
    Illegal AgeI site found at 1773
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 332


[edit]
Categories
Parameters
None