Difference between revisions of "Part:BBa K3328008"

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]

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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 32.3-fold due to the destruction of toehold hairpin structure.
+			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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