Part:BBa_K1711002
yeVenus-PEST-TheoA; Coding sequence for destabilized YFP Venus and theophylline-sensitive aptazyme
This part, yeVenus-PEST-TheoA, contains the coding sequence for a hybrid protein made of yeast enhanced yellow fluorescent protein (yeVenus) fused in frame with the CDS of the PEST-rich 178 c-terminal residues of Cln2, which targets the protein for ubiquitin dependent degradation. These coding sequences are fused with 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.
The time course comparison between the YFP-aptazyme and YFP-PEST-aptazyme 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.
PEST Reduces Background Fluorescence compared to non-PEST
Visual comparison of pellets under the dark reader after 4.5 hours between YFP-aptazyme and YFP-PEST-aptazyme at concentrations of theophylline 0 mM, 5 mM and 10 mM.
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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