Inspiration

5'UTR will be transcribed and can be used as a regulatory element to adjust the translation process. Thus, there is reason to believe that a delicate structure of the 5'UTR has the potential as a universal enhancement module. We see that Zhou et al. Screened some 5'UTR sequences from E. coli, which helped to improve protein content. Thus, we are prepared to consider this part as an enhancement module, the idea also received the author's support.[1]Zhou, Shenghu, et al. "Obtaining a Panel of Cascade Promoter-5′-UTR Complexesin Escherichia coli." Acs Synthetic Biology 6.6(2017)

Design

We selected J23108 from constitutive promoter family members and added the 5'UTR sequence thereafter. Fluorescent protein is used to characterize whether the 5'UTR has an enhanced effect on the promoter.

Proof of concept

We constructed two red fluorescent protein devices and constructed the loop by randomly selecting the promoter and the terminator. The difference is that UTRrpsT is added to a loop and the fluorescence intensity is measured using a microplate reader and plotted. We found that the fluorescence intensity of 6h increased by 1.5 fold. This indicates that UTRrpsT has the capability as a generic enhancement module.