Team:BS United China/Model

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Model

In our programs, our first basis is the basic growth curve of the bacteria that is involved in order to study the growth trend of the bacteria and to prepare for the analysis of the effectiveness of the inhibition of the engineering bacteria, along with proving that the existence of biology bricks would not affect the growth rate of the bacteria. The growth curve is the science of description of the growth trend of the bacteria based on measuring the intensity of the bacteria at same time intervals. These include E. coli，P. aeruginosa, Part one engineering, bacteria, and part two engineering bacteria.

The first step is to determine whether the engineering bacteria have a growth rate that differs from the growth rate of the E. coli. So, we added bacteria into the culture medium and measure the absorption of light every hour during the 16-hours duration.

Fig.1 The flow chart of the measure of the growth curve

The result is shown as the following graph. The curves of the engineering bacteria have similar shape as the curve of the E. coli. In the graph of the change of the growth rates, the trend of the rates of E. coli and engineering bacteria almost overlapped and reached the peak at the same hour at almost the same value.

Fig.2 The growth curve of the P. aeruginosa (Left top）The growth curve of the E. coil（Right top）The growth curve of the AHL scout（Left bottom）The growth curve of the AHL terminator（Right bottom）

Fig.3 Comparasion of the growth rates of the four kinds of bacteria

The second step is to measure the effectiveness of the part 1 engineering bacteria on the purpose of indicating the concentrations of P. aeruginosa. So the experiment uses the solution containing part 1 engineering bacteria and added the supernatant of P. aeruginosa into it. The result is that without supernatant, the solution of engineering bacteria produces much less fluorescence than the solutions with supernatant added. So the engineering bacteria is useful when indicating the present of P. aeruginosa.

Fig.4 The flow chart of the bacteria purification

Table 1

Then is to build the relationship between the light intensity between the AHL concentration and the intensity of the bacteria. The experiment is to add decreasing concentration of the AHL solution into the container of part 1 engineering bacteria. The result is that the concentration of the intensity of the bacteria is strongly related with the AHL concentration and the formula is y=151312x+4E+07

Table 2.The intensity of the intensity and the bacteria to the AHL concentration

Fig.5 The standard curve of the intensity of the bacteria to the AHL concentration

The next step is to measure the effectiveness of the PVDQ proteins on the reduction of the P. aeruginosa. The experiment is to add PVDQ proteins produced by part 2 engineering bacteria into the solution containing P. aeruginosa and measure the absorption value and calculate the intensity of the bacteria.

Fig.6 The flow chart of the inhibition of PVDQ protein on the bacteria

The result is that the PVDQ protein is effective during the first hour but the intensity increases in the second hour and the proteins reduced at first wash is most effective. Taking the fact of the bacteria intensity of the control group also decreased during the first hour, the rate of the reduction of the bacteria intensity is -1215037 per hour.

From the graphs of 8 to 11, the result shows that the concentration is the highest when at first wash, and the inhibition effect of PVDQ at high concentrations is the strongest. Comparing the control group with the proteins from the 5th wash and the 8th wash, the gradient is relatively less. Hence, the proteins also have some inhibition effect. Therefore, the conclusion is that PVDQ has relatively well inhibition effects.

Fig.7 The change of bacteria intensity control group

Fig.8 The change of bacteria intensity after treated by high protein concentration(50ng/ul)

Fig.9 The change of bacteria intensity after treated by medium protein concentration(25ng/ul)

Fig.10 The change of bacteria intensity after treated by low protein concentration(5ng/ul)