Team:Fatih-Medical/DJcoli/Assembly

To achieve our goal that being able to amplify the first detection signal which is started with the transfection reaction of TEV protease, we need to transform a sender and a receiver quorum sensing device into E.coli colonies, separately. In our design, there are the detector bacteria responsible for the recognition of tumor cells in the media or blood and the amplifier cells which increase the first signal via producing the great amount of reporter proteins. This communication of the amplification “request” comes from the quorum sensing device.

In our model, we plan to use a ready receiver-sender quorum sensing cassette which is present in the Registry. (BBa_F1610 and BBa_F2620) These systems consists of Lux promoter system which involves Lux promoter (pLuxR), LuxR regulatory protein and LuxI, an enzyme coding gene sequence which ends with the synthesis of 12HSLO3 that is necessary for the induction of Lux promoter with the help of LuxR.

Similarly, we designed a new quorum sensing cassette by using Las system instead of Lux. Almost the same, Las system uses Las promoter which is activated with the presence of LasR regulatory protein and 6HSLO3, a chemical compound that is produced by an enzyme which LasI gene codes. (BBa_K772005 and BBa_K772006) Hopefully, the sender bacterium in our project will be activated by the release of LasR through detection device, which results the production of quorum sensing elements, AHLs, to induce the second amplification response in the receiver bacteria in the media.

Note that the sender device has no promoter in the upstream region because of considering that different constitutive promoter alternatives may be used via assembling with BioBrick Standards. Also, note that the receiver device ends with pLas, but no ribosome binding site.