| 초록 |
The Potential of cis-regulatory riboswitch has been highlighted for monitoring various intracellular molecules. Several parameters of biosensor should be adjusted for application in user-defined working condition. However, tuning the parameters such as dynamic range, fold activation, and operational range is not straightforward. The intricate relationship between sequence and functional phenotype hampers the intuitive engineering. Thereby, straightforward tuning options have been studied. Here, we demonstrate three strategies to optimize functional parameters with an example of artificial L-tryptophan riboswitch. We modulated the copy number of the RNAs by tuning the promoter strength and plasmid origin to optimize dynamic range and fold activation. Then, we applied tetA to amplify the response by tetracycline supplementation. Finally, we constructed riboswitch variant by adopting an aptamer that shows lower affinity to L-tryptophan to reposition the operational range. |
