Part:BBa_K3078200
SfGFP under the J23104 promoter.
Use a constitutive promoter J23104 (BBa_J23104), natural RBS (BBa_J64022), sfGFP coding region (BBa_K2541400), and a double terminator (BBa_B0014). The composite part is used to measure the strength of the promoter J23104.
We hope that when we use an unrepresentated promoter, we can easily and accurately get its relative strength in the already characterized promoter family.
Therefore, we constructed the promoter strength measurement vector BBa_K3078100 and characterized the promoter with a large intensity gradient of the Anderson promoter family, BBa_J23119, BBa_J23104, BBa_J23108, BBa_J23105, BBa_J23114, and it is expected to establish a standard curve with fluorescence intensity.
Figure 1. The fluorescence of each promoter changes with time (fluorescence / OD600). Different elements are connected to the same measurement carrier, cultured overnight, diluted to OD = 0.02 the next day, and the fluorescence at absorption wavelength 528nm was measured under excitation wavelength 485nm light.
Table 1. Reported relative expression and corrected relative expression. We calculated the relative fluorescence intensity of each promoter for it based on J23119.
The fluorescence data showed the intensity of BBa_J23104 and BBa_J23119 is much too high to constitute a linear relationship with the other three promoters. The reason of which might be the fusion genes influenced the relatve expression level of the promoter.
We invited JiangnanU_China to repeat, and the result of its characterization was not consistent with reported relative expression .Therefore, we speculated that the relative strength might be affected by experimental strains, plasmids and other factors. Under our experimental conditions, the relative strength of Anderson promoter was not consistent with reported relative expression.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 476
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |