In our system, we have 3 parts. 2 of them are there for checking whether our composite part works and the last part is the main part for the mechanism of our project.
Our first 2 parts are the IbpB - RFP complex and the ROSE - RFP complex.
Both of these parts are RNA Thermometers with RFP's attached at their end. The part primarily starts off with pVeg2 promoter. As RNA Thermometers also carry the function of RBS', they come after the promoter. Finally the part ends with RFP.
The IbpB - RFP and ROSE - RFP parts were used as a testing mechanism to check if the RNA Thermometers work. The bacteria that are transformed with these parts are supposed to vary in color according to the temperature. If the bacteria are cultured in LB Media, the color of the fluid and the color concentration show us if the RNA Thermometers work or not. If several cultures are grown under different temperature conditions, the color concentration varies between the cultures.
The ROSE - Urease composite part is the main part of our project. It is a part which starts off with the pVeg2 promoter, continues with the ROSE Thermometer as the RBS and is completed with the Urease A, B and C trio. Between Urease A and B and also Urease B and C, there is an insulator and RBS.
This part forms the backbone of our project as it is responsible for the cooling mechanism. The ROSE Thermometer in the part can be masked or unmasked according to the temperature. This allows the control of the synthesis of the Urease enzyme. Also this permits the cooling in even cold environment.
The insulator and RBS are placed in the part for the part to work properly. As a result of the design, the long base length Urease A enzyme is prevented from being translated during low temperatures. This saves the bacteria from continously producing the longer Urease A molecule and prevents the bacteria from shouldering too much synthesis.
Also check out our working part, IbpB - RFP here.
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