Team:ETH Zurich/Part Collection
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Overview
Our whole cancer sensor depends on the sensitivity of the promoter regulated by LldR. LldR is a repressor that binds to O1 and O2 and forms a loop, avoiding any further transcription. If the promoter is too strong it will supress the transcription even in the presene of lactate, which will result in a reduced sensitivity of our system. If the promoter is too weak, it can result in an increased number of false positives. To decide the optimal arrangement of our regulatory system, we designed nine promoters by rearranging O1, O2 and a promoter.
Regulatory system design |
BioBrick |
Comments |
| lldRO1-plldR-lldRO2 | ||
| J23100-lldRO1-lldRO2 | ||
| J23100-lldRO1-R2oDNA-lldRO2 | ||
| lldRO1-J23100-lldRO2 | ||
| J23117-lldRO1-lldRO2 | ||
| J23117-lldRO1-R2oDNA-lldRO2 | ||
| lldRO1-J23117-lldRO2 | ||
| J23118-lldRO1-lldRO2 | ||
| J23118-lldRO1-R2oDNA-lldRO2 | ||
| lldRO1-J23118-lldRO2 |
Nomenclature
lldRO1: operon O1 of lldR. lldRO2: operon O2 of lldR. R2oDNA: biologically neutral DNA.
The second step of our system is a luxR protein inhibited both by LldR and LacI. To do that, we designed a series of regulators in which we combined lldR and lacI operators to find the optimal path for our system.
Regulatory system design |
BioBrick |
Comments |
| plac-lacO | ||
| placUV5-lacO | ||
| lldRO1-plac-lacO-lldRO | ||
| lldRO1-placUV5-lacO-lldRO2 | ||
| placUV5-lacO-lldRO1-R2oDNA-lldRO2 |
Nomenclature
lldRO1: operon O1 of lldR. lldRO2: operon O2 of lldR. R2oDNA: biologically neutral DNA. plac: lac promoter. lacO: lac operon. placUV5: enhanced lac promoter with strong expression in the case of lack of activation by cAMP.