Part:BBa_K4351018
Glucose binding RNA aptamer 1 + RNA Mango
Biosensor for detecting glucose with fluorescent output by RNA Mango.
Fluorescence experiments were completed to measure glucose binding by the aptamer by combining the glucose biosensor with RNA Mango part (BBa_K4351010).
RNA was unfolded by heating to 95°C and then let cool to room temperature in the presence of 20 µM ligand. The fluorophore TOI was only added to the RNA just prior to recording the fluorescence spectra and was excited at 510 nm. The emission spectra was recorded between 520 - 600 nm and two scans were completed.
From our experiments, we concluded that the addition of glucose to the RNA biosensor mix does result in an increase in fluorescence but only under certain conditions (Figure 4A). To use this biosensor to measure glucose binding, it first needed to be unfolded by heating to a high temperature and then allowed to refold and cool in the presence of glucose. The TOI was then added just prior to measuring the fluorescence. Our hypothesis is that the RNA is quite conformationally stable and cannot unfold to bind to glucose without first being melted. TOI can also not be added first, as it has a low nanomolar affinity for RNA Mango and would therefore bias the RNA folding to form the RNA Mango structure regardless of whether glucose was present or not.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 98
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