Difference between revisions of "Part:BBa K2904030"

Revision as of 12:40, 17 October 2019

Modular Four U riboswitch

Design

Background of 2019 OUC-China's project——RiboLego

Due to context-dependent performance and limited dynamic range, the widespread application of riboswitches is currently restricted. By replacing its original ORF with a new one, the structure of an aptamer domain can be subtly disrupted, resulting in a loss of ligand response. So riboswitch is still not be considered as a ‘plug and play' device. To tackle these problems, our project focuses on a standardized design principle to be used for modular and tunable riboswitch. The modular riboswitch we defined consists of the original riboswitch, Stabilizer and Tuner. Stabilizer can protect the structure of riboswitch from damage while Tuner can reduce the expression probability of fusion protein and make improvement of riboswitch function.

The construction of this part

To validate our design guideline, we employed a repressing Btub riboswitch, which can regulate the expression of TonB-dependent vitamin B12 receptor BtuB by binding adenosylcobalamin in Escherichia coli.The first 150bp of BtuB was chosen as Stabilizer of Btub riboswitch because our docking matrix suggested that a normal riboswitch structure would be observed when using this length of Stabilizer. This part was modular Btub riboswitch containing the original riboswitch, Stabilizer and Tuner A.

Construction of this part

After constructing modular kinetic riboswitches, we then want to demonstrate that our guideline can also apply to the thermodynamic riboswitch. So we designed a modular Four U riboswitch containing the original Four U riboswitch, Stabilizer and Tuner A. Using docking matrix, we selected the first 132bp of RFP as Stabilizer because RFP can express under the control of Four U.

Sequence and Features