Difference between revisions of "Team:ETH Zurich/Part Collection"

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Revision as of 22:41, 28 August 2015

"What I cannot create I do not understand."
- Richard Feynmann

Part Collection

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 BBa_K822000 Original promoter found in E. coli, published in 2012 by NTNU Trondheim
J23100-lldRO1-lldRO2 BBa_K1847002
lldRO1-J23100-lldRO2 BBa_K1847007
J23117-lldRO1-lldRO2 BBa_K1847003
J23117-lldRO1-R2oDNA-lldRO2 BBa_K1847005
lldRO1-J23117-lldRO2 BBa_K1847008
J23118-lldRO1-lldRO2 BBa_K1847004
J23118-lldRO1-R2oDNA-lldRO2 BBa_K1847006
lldRO1-J23118-lldRO2 BBa_K1847009

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 BBa_K1847013
placUV5-lacO BBa_K1847014
lldRO1-plac-lacO-lldRO BBa_K1847010
lldRO1-placUV5-lacO-lldRO2 BBa_K1847011
placUV5-lacO-lldRO1-R2oDNA-lldRO2 BBa_K1847012

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.

Design of the parts

Characterization