Difference between revisions of "Part:BBa K3447106"
(→Characterization) |
|||
Line 8: | Line 8: | ||
===Characterization=== | ===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.<br> | 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.<br> | ||
− | 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. | + | 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. 1A, compared to the control group, the growth and reproduction of <i>E. coli</i> cells in the experimental group were repressed.<br> |
− | Apart from this, plate-spreading experiment was also conducted to demonstrate the repressive function of relE (Fig. | + | Apart from this, plate-spreading experiment was also conducted to demonstrate the repressive function of relE (Fig. 1B). The result suggests that relE has inhibitory effects on our engineered bacteria.<br> |
[[Image: RelE was induced to inhibit bacterial growth.jpg|thumb|center|500px|<b>Fig. 1 RelE was induced to inhibit bacterial growth.</b> (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 OD<i>600</i>. 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.]]<br> | [[Image: RelE was induced to inhibit bacterial growth.jpg|thumb|center|500px|<b>Fig. 1 RelE was induced to inhibit bacterial growth.</b> (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 OD<i>600</i>. 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.]]<br> | ||
+ | |||
==<b>Design</b>== | ==<b>Design</b>== | ||
===Design Notes=== | ===Design Notes=== |
Latest 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. 1A, 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. 1B). The result suggests that relE has inhibitory effects on our engineered bacteria.
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 985