Part:BBa_K5226051
TcI42-T89
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Introduction
One of the goals of iGEM SCUT-China-A is to use synthetic biology tools to build thermosensitive bio-switch for Halomonas TD. We successfully built one using ci857 regulatory protein and superfolder green fluorescence protein(sfGFP). For better performance and broader range of usage, we mutated CI857. Because Halomonas TD grew at 30°C in a mediocre manner, we sought out a narrower temperature region, the CI857 variant, TCI-42, and mutated it as well. We characterized the mutants together with PR-sfgfp and several version of well-performed T-switch were successfully obtained.
Usage and Biology
T89 is a mutant of TCI-42,a widely used mutant of cI from bacteriophage λ as a thermogenetic tool. At 30℃ ,The cI857 dimer repressor binds in a cooperative form, by protein-protein interactions of their C-terminal domains that exhibits strong repression on R promoter. At 37℃, the dimer dissociates, So the R promoter works. TCI is a temperature-sensitive variant of the bacteriophage λ repressor cI. It consists of N-terminal, C-terminal and the linkage fragment between them. It is a cold-inducible transcription factor. When the temperature is below 35℃, its activity will gradually rising. When the temperature is above 35℃, its activity will remain at a low level. After our verification by experiment, T89 has lower leakage and higher fold change than CI857.
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