Our team has successfully validated a newly designed RFP coding device (RFP2) that contains double RFP reporting system. Our designing and wet lab experiments not only proved that our part has properly and efficiently worked but also helped us improve one of the most used parts in the iGEM registry, BBa_J04450.
Results are vivid; our validated part that we named RFP2 is showing brighter fluorescence under UV light and giving more intense color (purplish-red) under visible light than the random RFP device.
RFP2 under UV light along with controls, as brighter and beautiful as neon!
RFP2 under visible light, shocking pink and yummy!
We successfully improved one of the most frequently used parts in iGEM; RFP coding device BBa_J04450. The idea itself was based on our IDDP theory; ligating 2 RFP devices together and inserting them both in a single plasmid might make the color more intense and decrease the expression time to a certain extent.
Above gel shows the bands of our miniprep-ed RFP2 plasmids that were directly transformed from the ligated product. As it was direct transformation, some of the self-ligated plasmids too were extracted.
For the projects like us in which a biosensor gives color indication upon interacting with a target molecule, this improvement holds massive importance. By decreasing the expression time one can detect target compounds much earlier.
Our engineered bacteria that contain double-RFP expressing plasmid showed visible difference in the color as well as fluorescence in just 5hours of incubation. Plasmid having single-RFP device started to change the color after seventeen hours!
The picture above further fortifies the improvement that we have done. Sub-cultures of the transformed colonies clearly differentiate RFP and RFP2; our improved part.