| 초록 |
In synthetic biology area, various riboswitch-based genetic devices have been developed and applied to diverse fields. Since riboswitches should have distinct dose-response curves to meet the requirements for different applications, they have been redesigned by rational approaches. However, rational approach is applicable only to few riboswitches. Therefore, we developed an evolutionary approach for riboswitch engineering. First, we obtained naringenin (NRN) aptamer library by systematic evolution of ligands by exponential enrichment (SELEX), and constructed NRN riboswitch library composed of the aptamer library, random linker sequences, selection marker gene (tetA), and reporter gene (sgfp). Then we performed enrichments by dual selection in low, medium and high NRN concentrations. After the dual selection, we measured the dose-response curves of the enriched NRN riboswitches, and half maximal effective concentration (EC50) of riboswitches were classified according to the NRN concentration of enrichments. In conclusion, we established a novel approach for construction of riboswitches that have desired dose-response curves from scratch. |
