Team:OUC-China/Project/Results

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Results

Magnetic Receiver

1. In “parts construction” We amplified bacterial ferritin ftnA and submitted it into Registry.

2. In “Mag-attraction test” we found that placing a magnet under the plate of mineralized E .coli in fluid culture, E .coli will gather at the edge of magnet. And developed this discovery into a easy measurement method for magnetism of bacteria. You can see more in Captor page.

3. In “Ferritin expression test” we successfully purified the ferritin and proved the successful expression of ferritin using SDS-PAGE.

4. In “Transmission Electron Microscopy (TEM) Test”, we observed block in E .coli harboring PT5-ftnA, and assumed that block is over expressed ferritin. However, this result can’t precisely prove successfully mineralized ferritin.

5. In “Native PAGE Test”, referenced to mineralization test in vitro we proved a powerful test for in vivo mineralization. And successfully verify the mineralized ferritin FtnA with iron core.

6. In “Magnetic Properties of Magthermo coli”, through SQUID test, we successfully proved that most particles in Magthermo coli are superparamagnetic particles, fulfill the requirements for heating.

7. In “Alternative magnetic field heating detection” , we used optical fiber thermometer to detect the effect of heating, but temperature didn’t show apparent difference.

8. We found out the defect of previous measurement to detect heating performance of nanoparticles in vivo.

9. In “valuing protein concentration per cell” we explored methods for valuation of protein concentration per cell using plate reader and gel imaging. However, this method doesn’t work well.

Thermosensitive regulator

1. In “RNA Thermometer-select the best assembly” ,we assembled 9 different circuits: 3 constitutive promoters * 3 RNAT.

2. In “RNA Thermometer-select the best assembly-Agar plate test ” , we tested the 9 circuits on agar, cultured in 3 different temperatures: 28 ℃, 37℃, 42℃. After comparing, we found that RNAT-FourU(BBa_K115002) worked best, especially under the control of Promoter J23119 and J23101.

3. In “RNA Thermometer-select the best assembly-liquid test ” , we tested the 9 circuits cultured in 5 different temperature: 28℃, 35℃, 37℃, 40℃ and 42℃. We found that results in liquid is quite different from results on agar:
Four U(K115002) under the control of J23101 works best, and Four U(K115002) under the control of J23119 works far weaker than J23101+K115002.

Fig.1. Fluorescence intensity for 9 circuits

4. In “RNA Thermometer- Measure RNAT under heat stress “ , we assembled 4 circuits( please see the circuits on Thermosensitive Regulator page ). And value it under 5 different temperatures: 28℃, 35℃, 37℃, 40℃ and 42℃. And successfully proved that fluorescence under heat stress will decrease. Here’s the results:

Fig.2. Fluorescence intensity of GFP under different temperature

We can see the Fluorescence intensity of GFP under the control of constitutive promoter begins to fall from 37℃.
However, the Fluorescence intensity of RFP under the control of RNAT begins to rise from 37℃.

Fig.3. Fluorescence intensity of RFP under different temperature

This indicate that the switch efficiency of RNAT is higher than only taking RFP as indicator can show.

5. In “Thermosensitive T7 RNA Polymerase”, we assembled 2 T7 expression device, under the control of PT7 with %4 activity and %21 activity. Here’s the results:

Fig.4. Fluorescence intensity of GFP under the control of 4% and 21% PT7

6. In “Thermosensitive T7 RNA Polymerase”, we tested the best induce concentration of arabinose, on agar and in liquid, here’s the results:

Fig.5. induce concentration test on agar

It’s obvious that the best induce concentration on agar is 2%.

Fig.6. induce concentration test in liquid

It’s obvious that the best induce concentration in liquid is 0.002%

7.We haven’t finish the circuits of thermosensitive T7 RNAP. Thus, there’s no more results for this section. But we really love this part, and will continue this work!