Team:NTHU Taiwan/Contribution
Overview
From the previous study, NTU-Taida found that pFadBA (BBa_K817002) can be induced 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 cells?
- Will higher fatty acid concentration induce more fluorescence?
- How strong is the promoter pFadBA?
Effect of Fatty acid to E. coli
We discussed the relationship between fatty acid and the growth of E. coli to check suitable conditions for future experiments. First, we checked whether fatty acid would affect our experiments. Second, we chose 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 reaches 25mM, it inhibits the growth of E. coli. We can notice that a low concentration of fatty acid is somehow beneficial for cell growth. Therefore, we chose concentrations lower than 10 mM as our highest concentration to induce the promoter in the following experiments.
Fatty Acid Inducing to E. coli
As mentioned in the previous description, pFadBA (BBa_K3040007) can be induced by fatty acid because of the repressor protein. We can observe that with higher fatty acid levels, 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 induces the promoter, and higher concentrations induce more fluorescence.
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.
(J023110 with relative strength of 0.33 and J023014 with relative strength of 0.72)
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