Part:BBa_K2969042
J23119-TCI42-pL(3)-pR(3)-mRFP
This part contains the coding gene of the thermo-sensitive transcription factor TCI42 under the constitutive promoter J23119 and a reporter gene RFP under the promoter of TCI42. When the temperature is below 35℃, TCI42 is active to repress the transcription of RFP so that there will be little fluorescence.
Characterization
In 2019, UCAS-China developed a collection of thermosensitive parts with high-performance, versatility and robustness. Based on TCI transcription factor family and TlpA family, we collected five TCI and TlpA mutants and used sfGFP as reporter to build some heat-inducible ON-switches, which can open gene expression under high temperature. To characterize them quantitatively, we first characterized the performance of them by flow cytometer in Top10 strain.
As shown in Figure 1, most of the transcription repressors show sharp thermal transitions, especially TCI and TCI42, with more than 100-fold induction within 10 degrees Celsius. Their impressive performances make them candidate parts for our further circuit design.
What’s more, we also tested these heat-inducible ON-switch in the chassis E.coli Nissle 1917, a probiotic with more than 100 years of medical application, their robustness give us more confidence in the stability and preciseness of our ark. The result is shown in Figure 2.
Reference
Piraner, D. I., Abedi, M. H., Moser, B. A., Lee-Gosselin, A., and Shapiro, M. G., Tunable thermal bioswitch for in vivo control of microbial therapeutics. DOI: 10.1038/NCHEMBIO.2233
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1658
Illegal AgeI site found at 1770 - 1000COMPATIBLE WITH RFC[1000]
| None |