Part:BBa_K2969043
J23119-TlpA36-pTlpA-mRFP
This part contains the coding gene of the thermo-sensitive transcription factor TlpA36 under the constitutive promoter J23119 and a reporter gene mRFP under the promoter of TlpA36 pTlpA. When the temperature is below the transition temperature of TlpA36, TlpA is active to repress the transcription of mRFP 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
Illegal NotI site found at 589 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 874
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1177
Illegal AgeI site found at 1926
Illegal AgeI site found at 2038 - 1000COMPATIBLE WITH RFC[1000]
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