Part:BBa_K3580003
Plux/tet-GFPssrA(LVA)
To see the improvement in degradation effect by ssrA tagging, we construct a improved part (BBa_K3580003) by modification of an existing part: Plux/tet-GFP(BBa_K934025).
In the behavior of the gene circuit, it is important to reduce the concentration of proteins by degradation and dilution as well as production. Although cells can decrease their concentration by dilution according to growth, cell-free systems can’t because it doesn’t growth. Therefore, an improvement is necessary to incorporate degradation into the cell-free system. In order to compare these parts(BBa_K3580003 and BBa_K934025), we first measured the fluorescence of GFP in vivo for 240 minutes. The fluorescence of tagged GFP(BBa_K3580003) was lower than that of normal GFP(BBa_K934025) at 240 min point (Fig 2). Although GFP is a stable protein with a β-barrel and much difficult to be degraded, this result shows that tagged GFP was successfully degraded as we desired.
Then, we compared the fluorescence of GFP in a cell-free system which was extracted from the E.coli containing luxR protein. Fig 3. is the result of the experiment. The fluorescence of tagged GFP(BBa_K3580003) couldn’t be measured because the value was too low. Those results show that ssrA tagged protein can be degraded much both in vivo and in vitro. Based on this data, we modified the degradation terms in the model.
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 745
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