Difference between revisions of "Part:BBa K3328002"

Latest revision as of 08:04, 27 October 2020

OFF-switch3

an OFF-switch to regulate the expression of downstream gene

Design

3WJ repressor switch RNA employs an unstable hairpin secondary structure. This unstable hairpin was previously demonstrated to be translationally active. When a complementary trigger RNA is expressed, the trigger will bind to the switch RNA, making the originally unstable 3WJ structure stable, and represses translation.

Result

In the validation of 3WJ, as with toehold, we also set blank control (IPTG=0 M, aTc=0 mg/mL). Compared with the group without trigger expression (IPTG=0.1 M, aTc=0 mg/mL), the group with trigger expression (IPTG=0.1 M, aTc=0.25 mg/mL) showed inhibitory effect. Error bar: SD (n=9).

Crosstalk was determined by dividing the arithmetic mean of the GFP fluorescence from a given trigger switch pair by the arithmetic mean of the GFP fluorescence for the cognate trigger switch interaction. GFP fluorescence was measured from n=9 biologically independent samples.

3WJ repressor showed remarkable ON/OFF ratio and had good orthogonality, we choose it as OFF switch. And we used it in the construction of logic gates and the design of some more complex circuits.

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]

@@ Line 15: / Line 15: @@
 https://2020.igem.org/wiki/images/thumb/c/c6/T--OUC-China--result_fig3.jpg/653px-T--OUC-China--result_fig3.jpg
-In the validation of 3WJ, as with toehold, we also set blank control (IPTG=0 M, aTc=0 mg/ml). Compared with the group without trigger expression (IPTG=0.1 M, aTc=0 mg/ml), the group with trigger expression (IPTG=0.1 M, aTc=0.25 mg/ml) showed inhibitory effect.
+In the validation of 3WJ, as with toehold, we also set blank control (IPTG=0 M, aTc=0 mg/mL). Compared with the group without trigger expression (IPTG=0.1 M, aTc=0 mg/mL), the group with trigger expression (IPTG=0.1 M, aTc=0.25 mg/mL) showed inhibitory effect. Error bar: SD (n=9).
 https://2020.igem.org/wiki/images/thumb/3/31/T--OUC-China--result_fig4.jpg/768px-T--OUC-China--result_fig4.jpg