Difference between revisions of "Part:BBa K3447106"

Revision as of 10:35, 27 October 2020

AHL induces endotoxin expression

This part can produce luxR protein constitutively, which constitutes the AHL sensing system. By connecting the relE gene downstream, the normal growth and reproduction of bacteria will be inhibited when AHL is present.

We use this part to sensing the presence of AHL, and activates downstream toxin expression to achieve the purpose of inhibiting the growth of bacteria in the presence of AHL.

Characterization

In order to verify the combined action of AHL and relE, we inserted relE in the downstream of Prlux, an AHL-induced promotor, and drew a growth curve to examine the expression of relE.
Therefore, to verify the inhibiting effect of relE, a series of AHL-containing bacteria supernatant were added into our engineered bacteria as our experimental group. According to Fig. 7A, compared to the control group, the growth and reproduction of E. coli cells in the experimental group were repressed.
Apart from this, plate-spreading experiment was also conducted to demonstrate the repressive function of relE (Fig. 7B). The result suggests that relE has inhibitory effects on our engineered bacteria.

Fig. 1 RelE was induced to inhibit bacterial growth. (A) AHL-containing bacteria supernatant was added into AHL-responding bacteria to induce the expression of relE. The whole system was incubated at 37℃. During the 20-hour incubation, the fluorescence intensity was measured every 1 hr at OD600. The experiment was performed three times in triplicate. ***, P < 0.001 from respective control using Student’s t test. (B) Plate-spreading test. On the left is our negative control while the right part was spread with 400 μL AHL-containing bacteria supernatant.

Design

Design Notes

We added some synonymous mutations to avoid part rules.

Source

We found this sequence data in GenBank.

References

Usage and Biology

Sequence and Features