Team:Worldshaper-XSHS/result.html
This is the Data for Functional analysis
Through data analysis, we get the following results and conclusions:
1.The growth status of E. coli DH5 α under different conditions
It can be seen from figure 1 that under the condition of sealing, the strain of Oxy1 grows slowly in 0-8 hours, and it increases rapidly at 8-10h.In the same condition, the strain of Oxy4 has been growing slowly, and was significantly different in 10 hours from the strain of Oxy1.When the sealing conditions were removed, the strain growth of Oxy4 was close to that of Oxy1.
Different plasmids have different effects on the growth of E. coli DH5 α.E. coli DH5 alpha, which was transformed by Oxy4 plasmids, is less likely to grow than that was transformed by Oxy1 plasmid. Compared with the sealed environment, the unsealed environment is more favorable for the growth of E. coli DH5 α transformed by Oxy4 plasmid.
Figure 1. The growth of
E. coli DH5 α
under different conditions
2.Comparison of dissolved oxygen concentration in different conditions
We set the oxygen concentration in the air to 100%, and the concentration of dissolved oxygen in the medium was detected by an oxygen electrode. As can be seen from figure 2, in the three groups of experiments, Oxy1 strain had the slowest consumption of oxygen, and the change of oxygen concentration in the two groups of Oxy4 was close.Combining data from figure 1, it can be seen in 8 h, though OD600 close to group 1 and group 2, but the dissolved oxygen concentration in the first group was obviously higher than that of group 2, so we guess Oxy4 plasmid can promote the existence of bacteria on oxygen consumption, to create a low oxygen environment.At 10 h, dissolved oxygen concentration in the 2 and 3 group was similar, but you can see from figure 1 that the growth of the third group was obviously higher than that of group 2. So we guess the E. coli DH5 α transformed by Oxy4 plasmid can grow better without seal, rapid growth of the bacteria causing the low oxygen environment of group 3.
Figure 2. Comparison of dissolved oxygen concentration in different conditions
3.Fluorescence intensity analysis at the excitation wavelength of 475nm
The fluorescence intensity of each group was measured at the excitation wavelength of 475nm(figure 3). As can be seen from the figure 3, the fluorescence intensity of “Oxy1 sealed” is higher than that of “Oxy4 sealed”, which we guess may be related to the number of bacteria (figure 2 shows the OD600 in “Oxy4 sealed” was lower). We compared the fluorescence intensity of the “Oxy1 sealed” and “Oxy4 unsealed”, and the “Oxy1 sealed” was better than the “Oxy4 unsealed” at 0h-6h, while lower than the “Oxy4 unsealed” at both 8h and 10h. We suspect that this is related to the concentration of the bacteria in the solution and the concentration of dissolved oxygen.Therefore, we analyzed the results after correction with the initial OD600, and plotted the correlation between the ajusted value and oxygen concentrations (figure 4, figure 5 and figure 6). As can be seen from figure 4, the Oxy1 strain expressed the strongest fluorescence expressi0n at ab0ut 7% oxygen concentration, and decreased when oxygen concentration reached 3%.However, with the decreased oxygen concentration, the expressi0n enhancement of the Oxy4 strain was obvious, and the relative fluorescence value was significantly higher than that of Oxy1 strain. From figure 6, we see that when the concentration of oxygen is lower than 1%, the expressi0n of GFP sudden increase. We suspect that the oxygen concentration threshold for Oxy4 GFP expressi0n was ab0ut 1%, of course this also need to be further experiment.
Figure 3 fluorescence intensity analysis at the excitation wavelength of 475nm
Figure 4. Fluorescence intensity of Oxy1 strain in sealed culture
Figure 5. Fluorescence intensity of Oxy4 strain in sealed culture
Figure 6. Fluorescence intensity of Oxy4 strain in unsealed culture
4.Fluorescence intensity analysis at the excitation wavelength of 395nm
395nm is another excitation wavelength of GFP, and we also tested it. The results are shown in figure 7. But the change of the fluorescence intensity is weaker than 475nm excitation light, and after the initial fluorescence value and OD600 of correction, the change of the fluorescence value trend is still not obvious, so we did not use it as a main reference data.
Figure7 fluorescence intensity analysis at the excitation wavelength of 395nm
Results
