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Applications of BBa_K747096
Characterization by UT-Tokyo iGEM 2014
During DNA assembly, we found that this promoter was activated in Escherichia coli. Then we decided to characterize this promoter activity by comparing with constitutive promoter (BBa_J23101) using GFP (BBa_E0040) (Figure 1). As seen in Figure 1, this promoter shows weaker activity than BBa_J23101.
In order to calculate RPU (relative promoter unit ), we performed real-time measurement of GFP fluorescence (Figure 2). We measured activity of GFP and OD600, which are necessary for calculation of RPU.
We calculated RPU(Relative Promoter Unit) of BBa_K747096 from the Fig. 2 using the equation (1).
This equation can be used only when Fluorescence per cell is in steady state. In this experiment, this condition was fulfilled as shown in Table1.
From Table1, we can get RPU of BBa_K747096: RPU = 5.8
Characterization by BIOSINT_Mexico iGEM 2014
CMV promoter fused with YFP by BIOSINT_Mexico iGEM 2014
As a part of the project from 2014_BIOSINT_Mexico , we fused the promoter PCMV (BBa_K747096) to a YFP reporter (BBa_E0030).
In order to measure its expression, we culture transformed E.coli in agar plates and measured the fluorescense intensity every 45 minutes. After that, we obtained the following data. [Fig.1]
We made three repetitions, so in order to model the equation, we obtained the mean measure, therefore:
We analysed the data and using the Wolfram Mathematica software, we obtained the following equation that fits the data. If we plot this equation, we obtain:
Where we can see that the intensity of YFP grows directly proportional to the time and concentration of the molecule.