Part:BBa_K1711001
yeVenus-TheoA; coding sequence for enhanced yellow fluorescent protein and theophylline-sensative ap
The part contains the coding sequence for yeast enhanced yellow fluorescent protein (yeVenus), linked to it is a gene for a theophylline sensitive riboswitch aptazyme. The aptazyme portion of the transcript self-cleaves in the absence of theophylline and no YFP should be produced. The theophylline bound state stabilizes the transcript, which translates to the protein and fluorescence should be observed.
Theophylline addition seems to cause a difference in fluorescence
Fluorescence comparison between 0 mM and 10 mM condition after 4 hour of incubation. construct expressed in yeast. Fluorescence is measured by flow cytometry in arbitrary units. 4 transformants tested and a negative control (no YFP expressing yeast). Transformant 2 has a point mutation T225I.
Comparison of aptazyme activity between 0 mM theophylline, 10 mM theophylline and 10 mM caffeine conditions. fluorescence measured every 1.5 hours.
Concentration assay. yeast incubated at 0 mM, 5 mM, 10 mM and 30 mM theophylline and negative control (no YFP). Fluorescence measured after 4 hours.
Theophylline Induction is Visible on a Macro Scale
Pellets from the concentration assay after the incubation period. pellets are from 0 mM and 30 mM theophylline condition. Visualized on dark reader.
Fluorescence and visual comparison of constitutively expressing YFP yeast (positive) and no YFP yeast (negative) over the period of 4.5 hours at concentrations of theophylline: 0 mM, 5 mM and 10 mM. Fluorescence measured with a flow cytometer every 1.5 hours in arbitrary fluorescence units. Note: 0 mM negative data might be due to human error. Pellets are from after 4.5 hours.
- 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 644
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