Difference between revisions of "Part:BBa K649105:Experience"
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[[Image:Odlsrr.png|thumb|center|500px|After four hours from OD590 reaching 0.15, we measured OD.<br>This work is done by Hiroki Yoshise.]] | [[Image:Odlsrr.png|thumb|center|500px|After four hours from OD590 reaching 0.15, we measured OD.<br>This work is done by Hiroki Yoshise.]] | ||
| − | [[Image: | + | [[Image:Lsrr.png|thumb|center|500px|Fluorescence intensity is decreased by LsrR repression.<br>This work is done by Hiroki Yoshise.]] |
Revision as of 23:26, 3 October 2011
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how you used this part and how it worked out.
Applications of BBa_K649105
We confirmed that LsrR represses lsrA promoter. We measured LsrR repression activity by using a plasmid (BBa_K649105) which have a lsrR gene downstream of lsrA promoter-gfp.By the LsrR repression, the fluorescence intensity decreased 3-fold. This result shows that LsrR successfully repressed lsrA promoter. The working part we created allow the use of AI-2 as a signaling molecule, which is a very powerful tool to build complex Synthetic Biology systems.
[Sample]
Ptet-gfp on pSB6A1(JM2.300)(positive control)
Promoterless-gfp on pSB6A1(JM2.300)(negative control)
PlsrA-gfp on pSB3K3(MG1655)
PlsrA-gfp-PlsrR-lsrR on pSB3K3(MG1655)
[Method]
1. Overnight cultures of reporter strains grown at 37 °C in LB medium containing appropriate antibiotics were diluted 1:100 into 3 ml of LB medium and were incubated at 37 °C as fresh cultures.
2. After their OD590 reached 0.15, the fresh cultures were diluted 1:10 or 1:100.
3. After 4-hour incubation at 37 °C, 1 ml of each culture was moved to 1.6ml tube and its fluorescence intensity was measured with a flow cytometer.
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