Difference between revisions of "Part:BBa K4169011"

Revision as of 11:16, 12 October 2022

A RNA thermometer mutation 2

Based on the characteristics of secondary structure changes of RNA under different temperature conditions, the RNA thermometer element was introduced in this project. An RC sequence is present on this RNA thermometer, which recruits RNase E to degrade subsequent RNA fragments. At low temperatures,such as 27℃, RNA thermometers will maintain the secondary stem-loop structure, and RC sequences will complement and pair with other bases, unable to recruit RNase, so subsequent sequences will be expressed normally. However, at 37℃, the secondary structure opens, and the subsequent RNA fragments are cleaved and degraded by RNA enzymes.In order to change the temperature at which it functions, we mutated its gene sequence, that is, the first A after ARC was changed to T.

Design and Mutation

We designed the mutation primers (Figure 1.)and the expected mutation sequences(Figure 2.)