Part:BBa_K4988001
oleic acid promoter
Oleic acid can effectively inhibit the formation of cholesterol [1] and oleic acid is very common in food, because oleic acid can degrade low-density cholesterol, and oleic acid in the form of glyceric acid is in large quantities in animal and plant oils. Moreover, oleic acid has little effect on bacterial growth and can activate the expression of oleic acid promoter. As a result,we decided to use oleic acid promoter as our promoter.
Usage and Biology
We used oleic acid promoter as our promoter, which can be activated by oleic acid, to stimulate the expression of dowstream gene and transform the plasmids into E.coli Rosetta.
Figure 1 Design of the Oleic acid.
Characterization
The LB medium was added with different concentrations of oleic acid and inoculated with E.coli Rosetta. The OD600 value of E.coli Rosetta was measured by spectrophotometer. The results as figure 2A showed that 5 mM oleic acid had little effect on bacterial growth, while 10 mM oleic acid inhibited bacterial growth.
The LB medium was added with different concentrations of oleic acid and inoculated with E.coli Rosetta/pOleic ACID-MRFP. Adjust the initial OD600 to 0.1. After 12 hours, the OD600 value and mRFP fluorescence intensity were measured using an microplate reader. The results showed that under the induction of oleic acid at 0mM and 5 mM, oleic acid promoter mediated the engineering bacteria to produce about 4.5 Fold change under the induction of 5mM oleic acid comparing to the result produced by omM oleic acid in figure 2B.
Figure2：Effect of oleic acid on bacterial growth and response of oleic acid promoter.
Potential application directions
Experiments shows that oleic acid can effectively activate the oleic acid promoter which can further stimulate the expression of downstream gene. As a result, oleic acid promoter is going to be useful in producing low cholesterol foods and helps decrease the cholesterol intake of potential high cholesterol patients and helps decrease the risk of diseases such as cardiovascular disease caused by high cholesterol. Oleic acid promoter can be used as a clinical treatment for high cholesterol in the future and also provide new ideas and solutions.
References
1. Priore P, Gnoni A, Natali F, Testini M, Gnoni GV, Siculella L, Damiano F. Oleic Acid and Hydroxytyrosol Inhibit Cholesterol and Fatty Acid Synthesis in C6 Glioma Cells. Oxid Med Cell Longev. 2017;2017:9076052. doi: 10.1155/2017/9076052. Epub 2017 Dec 24. PMID: 29435099; PMCID: PMC5757140.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |