| 초록 |
The potential of RNA encoded sensors which exploits the mechanism of translation initiation has been highlighted for monitoring metabolically significant biomolecules in vivo. Several parameters, such as operational range, fold activation and dynamic range, of the sensor should meet the requirements to be applied in user-defined condition. However, tuning parameters of riboswitch is not straightforward since those are dependent on the conformation of the template RNA. Therefore, it is necessary to develop alternative simple tuning options. Here, we demonstrate three strategies to modulate them by engineering Trp riboswitch for example. We modulated the strength of the promoter or the copy number of the plasmid to optimize dynamic range and fold activation. Then tetA was utilized to amplify the response by antibiotics supplementation. Finally, to moidfy the operational range, a riboswitch variant was constructed using an aptamer with distinct Km value to L-Trp. |
