Difference between revisions of "Part:BBa K3328008"

 
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an ON-switch to regulate the expression of the downstream gene
 
an ON-switch to regulate the expression of the downstream gene
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===Design===
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Toehold switch is composed of cis-acting element RNA hairpins and trans-acting factor trigger RNA. The binding of a trigger RNA to the toehold sequence allows for a branch migration process, exposing AUG and RBS for translation initiation.
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===Result===
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<html>
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<head>
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<meta charset="utf-8">
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</head>
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<body>
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<div>
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<img src="https://2020.igem.org/wiki/images/7/7e/T--OUC-China--result_fig2_th.jpg" style="width: 80%;">
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</div>
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<p>
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Compared with the blank control (IPTG=0 M, aTc=0 mg/mL), the fluorescence of GFP was low when only the promoter before toehold sequence was turned on (IPTG=0.1 M, aTc=0 mg/mL). This indicates that the toehold has the advantage of low leakage. When the trigger was expressed (IPTG=0.1 M, aTc=0.25 mg/mL), it showed a high GFP fluorescence of up to 32-fold due to the destruction of toehold hairpin structure. Error bar: SD (n=9).
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</p>
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</body>
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</html>
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===Usage and Biology===
 
  
 
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Latest revision as of 08:50, 27 October 2020


ON-switch

an ON-switch to regulate the expression of the downstream gene

Design

Toehold switch is composed of cis-acting element RNA hairpins and trans-acting factor trigger RNA. The binding of a trigger RNA to the toehold sequence allows for a branch migration process, exposing AUG and RBS for translation initiation.

Result

Compared with the blank control (IPTG=0 M, aTc=0 mg/mL), the fluorescence of GFP was low when only the promoter before toehold sequence was turned on (IPTG=0.1 M, aTc=0 mg/mL). This indicates that the toehold has the advantage of low leakage. When the trigger was expressed (IPTG=0.1 M, aTc=0.25 mg/mL), it showed a high GFP fluorescence of up to 32-fold due to the destruction of toehold hairpin structure. Error bar: SD (n=9).



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]