Part:BBa_M36708:Design
Pyruvate Sensor
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 679
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We chose to use a medium level 5' UTR meaning the ribosome binds the DNA roughly 50% of the time because IclR inhibits the aceBAK promoter without pyruvate so having extremely high levels of IclR would mean that regardless of the pyruvate concentrations, the aceBAK promoter will always be inhibited. With IclR at a medium level in the cell, the presence of pyruvate should stabilize the ability of IclR to bind the DNA and inhibit the ribosome binding.
We chose to use the aceBAK promoter and IclR because of the relationship of pyruvate to IclR and the aceBAK promoter. The latter half of the experiment is the relationship between pressure and the expression of the test actuator downstream of the aceBAK promoter. Lactate Dehydrogenase, LDH, converts lactate into pyruvate under aerobic, non-acidic conditions. Pressure inhibits the activity of LDH so less pyruvate was present in the cell so the aceBAK promoter is less inhibited so there are higher levels of the test actuator downstream of the aceBAK promoter in the cell.
Source
Katie Lund and Wyatt Woodson parts obtained from Stanford BIOE44 - S11