We constructed 5 plasmids [positive control, negative control, device1~3] by using BioBricks and then transformed to E. coli K-12 JM109. We checked them by colony direct PCR and plasmids sequence. We cultivated correct bacterias by shaking until OD660:0.5. Cultivated mediums were transferred to 96-well plate by 100mL. We read the absorbance.
Setting of plate reader is following:
Sequence data
・device 1 : J23101+ I13504
・device 2 : J23106+ I13504
・device 3 : J23117+ I13504
These sequence datas are correct.
Result
Definition
We expressed absorbance as relative value for positive control. This expression is following:
Sample : Absorbance in sample
NCav : Average of absorbance in negative control
PCav : Average of absorbance in positive control
Value
Relative absorbance values for positive control are shown below.
The value of device1 is much higher than that of device2 and 3. This is why each device is different from promoter sequence, especially -10 and -35 region.
Promoter sequence in device1 resembles consensus sequence. On the other hand, device2 and 3 is not similar than device1. This is how each GFP expression is different.
