Part:BBa_K1892004:Experience

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Applications of BBa_K1892004

We demonstrate that our project is effective in LL-37 and the motor protein yebf through the experiments below. Antimicrobial Effect of LL-37: To test the wide anti-bacterial function of LL-37 expressed by E.coli, we selected three commonly found representative bacteria including both Gram-positive and Gram-negative Bacteria: Staphylococcus aureus, Bacillus subtilis, and the not genetically modified Escherichia coli, using the LB culture for cultivating the bacteria. First, we spread the liquid bacteria on the LB solid in petri dish. After the film of liquid bacteria is completely dry on the LB solid culture, we separate the petri dish into four parts as the diagram below shows.

Figure 1: diagram for experiment design

In this experiment, water is the negative control and antibiotic carbenicillin as the positive control. On each section, we add a drop of 10μL of the corresponding liquid. After all the liquid is dry, the petri dishes are placed in 37℃ incubators for cultivation. We set up comparisons as well, one section is added with induced yebf-LL-37 as the experiment group; another one is added with yebf-LL-37 that had not been induced as the control group.

The results:

Figure 1. Antimicrobial effect of yebf-LL-37 against Bacillus subtilis

Figure 2. Antimicrobial effect of yebf-LL-37 against Bacillus subtilis

Figure 3. Antimicrobial effect of yebf-LL-37 against E.coli

The results matched our expectation and proved our antimicrobial concept. For all three types of bacteria, yebf-LL-37 that had not been induced did not produce an antibacterial effect while ampicillin is effective. For the part added induced yebf-LL-37, we can see a clear circle in the middle of each section with induced E.coli, which mean that LL-37 effectively inhibited bacterial growth in B.subtilis and S.aureus. The effect of it in E.coli is not so obvious, which may due to that the chassis is E.coli. From this experiments, we proved that the antimicrobial peptide LL-37 is effective against both Gram positive and Gram-negative bacteria.

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