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Team NTHU Taiwan 2019: Characterization of effects and inducing of fatty acid to E.coli and Relative Fluorescence to Anderson Promoter
Purpose
From previous study, NTU-Taida found that pFadBA can be induce by adding fatty acid. We’d like to understand more about this promoter by answering the following questions:
- Does fatty acid affect the growth of cell?
- Will higher fatty acid concentration induce more fluorescence?
- How strong is the promoter pFadBA?
Effect of Fatty acid to E. coli
We discuss the relationship between fatty acid and the growth of E. coli to check suitable conditions for future experiments. First, we check whether fatty acid affect our experiments. Second, we choose the best concentration for future experiments.
In general, relative cell concentration does not change very much. It indicates that under 0.4mM and 3.125mM fatty acid, E. coli might grow better than control. However, when the concentration is 25mM, it inhibits growth of E. coli. We can notice that low concentration of fatty acid is somehow benefit for cell growth. Therefore, we choose concentration lower than 10 mM as our highest concentration to induce the promoter in the following experiments.
Figure 1. Fluorescence detection in 96-well
Figure 2. Relative cell number with the different concentration over time
Fatty Acid Inducing to E. coli
As our previous description, pFadBA can be induced by fatty acid because of the repressor protein. We can obviously observe that with higher fatty acid level, the fluorescence level will rise. The fold change is almost 2.5 times in 0.625 mM LB, but is only 1.7 times in 0.156 mM LB. This data shows that fatty acid actually induce the promoter, and higher concentration induces more fluorescence.
Figure 3. Relative fluorescence of cell treated with different fatty acid concentration over time
Relative Fluorescence to Anderson Promoter
To know the intensity level of pFadBA expression, we compare the intensity between pFadBA and Anderson promoter. By comparing pFadBA with Anderson promoter, it demonstrates that the fluorescence level of pFadBA is much lower than the J023104 and approximately 10 percent level of J23110.
Figure 4. Comparison of pFadBA with the Anderson promoter
(J023110 with relative strength of 0.33 and J023014 with relative strength of 0.72)
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