The Design of a 5-HTP Biosensor with Fluorescence Detection by Combination of a 5-HTP Detection Aptamer and Fluorescent Malachite Green Aptamer
McGinnis, Courtney D.
Non-coding RNA can be evolutionarily selected based on their ability to associate their tertiary structure to a small molecule with a high affinity. Base pairs of the RNA molecules specifically involved in the association of a small molecule can be pieced together with other RNA segments to form aptazyme. In the experimental design of an aptazyme to detect 5-HTP with MG detection, three RNA segments were spliced together. These three segments were a 5-HTP ligand-binding region, connected by communication module to the fluorophore aptamer. Each section was treated as a unit and modified based on the modeled folding of the RNA sequences. DNA complement sequences of four modeled biosensors were made and RNA translation was optimized. The flourophore malachite green was characterized based on in the fluorescence to most accurately measured between 1 – 10 μM.
viii, 11 p.
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