Part:BBa_K4785005:Experience

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Applications of BBa_K4785005

1.Protein function verification

In the inhibition of biofilm growth experiment (Fig.1), we can observe that PslG(BBa_K4785004) treatment group effectively inhibited the growth of biofilm by comparing to the control group. HMGB1-A box shows the strongest ability to inhibit the biofilm growth among four HMGB1 modified proteins.

Figure 1:

In the biofilm disassembly experiment (Fig.2), the PslG treatment group reduced the OD550 absorption value corresponding to the biofilm biomass from about 1.5 to about 0.5 in the first two hours, while HMGB1-A box shows the strongest ability to disassemble the biofilm among four HMGB1 modified proteins.

Figure 2:

According to those results, we decided to choose PslG and HMGB1-A box as our functional proteins.

2.Intuitive observation bvconfocal microscopy

After co-treatment with HMGB1-A box and PslG for 24h, we imaged biofilm by confocal microscopy. First, we observed that in the control group, a large area of intact biofilm structure appeared, and eDNA and Psl had good colocalization. In the experimental group, most of the biofilm was disrupted, the grid structure was destroyed, and most of the eDNA and Psl were degraded by staining (Fig.3). According to the thickness of the residual biofilm, the thickness of the treated biofilm was also significantly reduced (Fig.4). These results all illustrate more intuitively the role of our target proteins.

Figure 3:

Figure 4:

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