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Revision as of 03:12, 1 November 2017
Result
We will show our results from three aspects:
Qualitative analysis:After 12 hours of incubation(LB,37℃,200r/min), we observed the bacteria on a fluorescence microscope.
In the medium with the addition of nickel ions, we can see visible red fluorescence.
![this is a photo](https://static.igem.org/mediawiki/2017/5/50/Hbut-result-1.png)
In other media, there is no such phenomenon. The whole vision is black.
![this is a photo](https://static.igem.org/mediawiki/2017/4/48/Hbut-result-2.png)
![this is a photo](https://static.igem.org/mediawiki/2017/e/e5/Hbut-result-3.png)
![this is a photo](https://static.igem.org/mediawiki/2017/a/ab/Hbut-result-4.png)
![this is a photo](https://static.igem.org/mediawiki/2017/3/3d/Hbut-result-5.png)
![this is a photo](https://static.igem.org/mediawiki/2017/1/1f/Hbut-result-6.png)
So our device only responds to nickel ions.
Quantitative analysis:The initial induction concentration was OD0.8, and the induction time was 2 hours. Under these conditions, the nickel ion concentration had a significant fucntional relationship with the fluorescence itensity.
Especially in the nickel ion concentration of 10-6mmol / L to 10-4mmol / L between.
![this is a photo](https://static.igem.org/mediawiki/2017/7/71/Hbut-result-7.png)
At first, we made some assumptions.
Assumptions:The degradation of and other substances are linear.
Every cell has the same state.
The reaction in the process satisifies the requirements of the first-order kinetic reaction.
Then we reached some conclusions through modeling.
Incomplete Repression:Under incomplete repression, the bacteria express red fluorescent proteins without the presence of nickel ions.
The incomplete repression of promoters is a major issue in our fluorescence system. This issue was particularly observed in following diagram.
![this is a photo](https://static.igem.org/mediawiki/2017/6/60/8.png)
The solution of this differential equation is as follows:
![this is a photo](https://static.igem.org/mediawiki/2017/a/a1/Hbut-result-9.png)
Or experiment is designed to induce the expression fo fluorescent proteins in the presence of nickel ions, however, we found that above a certain concentration of nickel ions, the expression of fluorescent proteins will be reduced due to nickel ion cytotoxicity.
The July 20 experimental data is as follows:
![this is a photo](https://static.igem.org/mediawiki/2017/7/7b/Hbut-result-10.png)
The different equation of the red fluorescent protein is then modified to:
![this is a photo](https://static.igem.org/mediawiki/2017/a/a5/Hbut-result-11.png)
Where
![this is a photo](https://static.igem.org/mediawiki/2017/e/e2/Hbut-result-12.png)