Part:BBa_K1682018
PLTet0-1:NarL, PdcuS:sfGFP, J23109:TetR
Functional nitrate anion sensor based on the NarX/NarL Two-Component Signal Transduction System in E. coli with semi-tunable dynamic range and sensitivity. NarL expression is regulated by the TetR-PLTet0-1 1-component regulatory system, whereby addition of ATC (anhydrotetracycline) induces higher NarL expression. This regulates the ratio of native NarX/NarL protein in the cell, controlling to an extent the dynamic range and sensitivities of the sensor. Phosphorylated NarL binds to the repressible PdcuS promoter from E. coli, regulating the expression of sfGFP. Low nitrate conditions result in high sfGFP expression, high nitrate conditions repress transcription. The sensitivity of the sensor is in the sub-mM to mM range for nitrate. Further testing is needed to fully characterize the relationship of ATC-induction to nitrate sensitivity and dynamic range. The sensor is functional in the absence of ATC with a low dynamic range, and ATC concentrations of 1-10 ng/ml increase dynamic range and sensitivity. Further increase of ATC drives the ON-state down, decreasing dynamic range.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2037
Illegal NheI site found at 2060 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 318
Illegal BglII site found at 486
Illegal BamHI site found at 1995
Illegal XhoI site found at 31 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1149
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