Difference between revisions of "Part:BBa K5226050"
 
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===Introduction===
 
===Introduction===
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One of the goals of iGEM SCUT-China-A is to use synthetic biology tools to <b>build thermosensitive bio-switch</b> for <i>Halomonas</i> TD.We successfully built one <b>using ci857 regulatory protein</b>. And for better performance and broader range of usage, we <b>mutated CI857 and its mutant TCI-42 to see whether we could get one or several well-performed mutant with lower leakage and higher fold change</b>.
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One of the goals of iGEM SCUT-China-A is to use synthetic biology tools to <b>build thermosensitive bio-switch</b> for <i>Halomonas</i> TD. We successfully built one <b>using ci857 regulatory protein and superfolder green fluorescence protein(sfGFP)</b>. For better performance and broader range of usage, we mutated CI857. Because <i>Halomonas</i> TD grew at 30°C in a mediocre manner, we sought out <b>a narrower temperature region, the CI857 variant, TCI-42, and mutated it as well</b>. We characterized the mutants together with PR-sfgfp and several version of well-performed T-switch were successfully obtained.
  
 
===Usage and Biology===
 
===Usage and Biology===
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T88 is <b>a mutant of TCI-42</b>,a widely used mutant of cI from <i>bacteriophage</i> λ as a thermogenetic tool. <b>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, C27 has lower leakage and higher fold change than CI857.</b>
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T88 is <b>a mutant of TCI-42</b>,a widely used mutant of cI from <i>bacteriophage</i> λ as a thermogenetic tool. <b>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 <b>temperature-sensitive variant</b> of the <i>bacteriophage</i> λ repressor cI. It consists of <b>N-terminal, C-terminal and the linkage fragment</b> between them. It is a <b>cold-inducible transcription factor</b>. When the temperature is <b>below 35℃</b>, its activity will <b>gradually rising</b>. When the temperature is <b>above 35℃</b>, its activity will <b>remain at a low level</b>. After our verification by experiment, <b>T88 has lower leakage and higher fold change than CI857.</b>
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Latest revision as of 03:25, 30 September 2024


TcI42-T88

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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

T88 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 <b>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, T88 has lower leakage and higher fold change than CI857.