Part:BBa_K5226047
CI857-C34
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
C34 is a mutant of CI857,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.After our verification by experiment, C34 has lower leakage and higher fold change than CI857.
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