Team:BFSUICC-China/Results

Construct Parts

Circuit

We use the elcetrophoresis to detect whether the parts connect successfully.
The right column is the marker and the strips on the left are the elcetrophoresis of BBa_K 2555004. As shown in the diagram below, our parts has been ligated successfully. The process is achieved by digestion and ligation to pSB4K5 and pSB1C3.

Sensor Part

We change the backbone of BBa_K2555000 in pUC 57 to pSB4K5 to pSB1C3. The right column is the marker, and the two strips on the left are restriction digestion products of BBa_K2555000. The sizes of the two strips are both about 1128 bp.

Regulator Part

We change the backbone of BBa_K2555002 in pUC 57 to pSB4K5 and pSB1C3. The right column is the marker, and the two strips on the left are registriction digestion products of BBa_K2555002. The sizes of the two strips are both about 1819 bp.

Test the function of parts

Circuit

Test the function of BBa_K2555004

The results of a plate reader experiment with different arabinose concentration after five hours

Find the biggest dynamic range of arabinose
The results of dynamic range of BBa_K2555004 with different arabinose concentrations after five hours.

Dynamic range
(highest value(concentration of copper ions is 2.048mM)-lowest value(concentration of copper ions is 0))/(lowest value(concentration of copper ions is 0))
Under o.1 mM arabinose BBa_K2555004 has the biggest dynamic range, so the amount of CueR under o.1mM arabinose may has positive effect on activation of pCop A.

Compare the function of BBa_K15550000 and BBa_K2555004

The results of a plate reader experiment with different copper and arabinose concentrations after five hours.
Fluorescent increase rate=(output（high）-output(low))/(input(high)-input(low))

The relative fluorescence intensity, the increasing rate of relative fluorescence intensity and dynamic range of BBa_K2555004 under 0.1 mM Arabinose were all significantly higher than that of BBa_k1555000.

Sensor part

Compare the function of BBa_K25550000 and BBa_K1555000
The results of a plate reader experiment with different copper concentrations after five hours.

We optimized BBa_k1555000 to be BBa_K2555000. The relative fluorescence intensity of BBa_K2555000 under different copper ion concentrations was higher than that of BBa_k1555000, and the increasing rate of relative fluorescence intensity was higher than that of BBa_k1555000.

Regulator part

Test the expression of CueR

The results of a OD600 with different arabinose concentrations after five hours.

With the increase of Arabinose concentration, the relative fluorescence value of the construct increased, which indirectly reflected the CueR protein expression of BBa_K2555002 increased with the increase of Arabinose concentration.

Test the effect of Cue R expression on growth of bacteria.