Part:BBa_K3328000:Experience

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Applications of BBa_K3328000

Lower Leakage and Higher ON/OFF Ratio

Our experiments were done with both the improved part (BBa_K3328000) and the original part (BBa_K2150031) as a control. Our part has lower leakage and higher ON/OFF ratio than the T7 promoter.

In the blank control (IPTG=0 M), the fluorescence of T7 promoter (BBa_K2150031) is higher than 3WJ Switch1 (BBa_K3328000). This indicates that our part (BBa_K3328000) has the advantage of low leakage. When the inducer is added (IPTG=0.1 M), 3WJ Switch1 showed a high ON/OFF ratio of up to 15-fold, and the T7 promoter has the ON/OFF ratio of only 3-fold.

The OFF-switch

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.

NOT gate

The left figure shows that when INPUT=0, the fluorescence of GFP is high. And the fluorescence intensity of GFP was low when INPUT=1. This corresponds to the situation described in the truth table on the right. INPUT=1 means that aTc (0.25 mg/mL) is added. Error bar: SD (n=9).

IMPLY gate

The left figure shows that when INPUT A=0, INPUT B=1, the fluorescence of GFP is low. And the fluorescence intensity of GFP was high in the other three groups. This corresponds to the situation described in the truth table on the right. INPUT A=1 means that aTc (0.25 mg/mL) is added, INPUT B=1 means that HSL (0.1 mg/mL) is added. Error bar: SD (n=9).

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