Difference between revisions of "Part:BBa K649105:Experience"
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===Applications of BBa_K649105=== | ===Applications of BBa_K649105=== | ||
| − | We characterized BBa_K649105 E.coli MG1655. | + | We characterized BBa_K649105 in <i>E.coli</i> MG1655. |
| − | [[Image: | + | [[Image:LsrRBA.png|thumb|center|500px|Fluorescence intensity is decreased by LsrR repression.<br>This work is done by Hiroki Yoshise.]] |
| − | 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. | + | We confirmed that LsrR represses lsrA promoter. We measured LsrR repression activity by using a plasmid (BBa_K649105) which have a <i>lsrR</i> gene downstream of lsrA promoter-<i>gfp</i>.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] | [Sample] | ||
| − | Ptet-gfp on pSB6A1(JM2.300)(positive control) | + | Ptet-<i>gfp</i> on pSB6A1(JM2.300)(positive control) |
| − | Promoterless-gfp on pSB6A1(JM2.300)(negative control) | + | Promoterless-<i>gfp</i> on pSB6A1(JM2.300)(negative control) |
| − | PlsrA-gfp on pSB3K3(MG1655) | + | PlsrA-<i>gfp</i> on pSB3K3(MG1655) |
| − | PlsrA-gfp-PlsrR-lsrR on pSB3K3(MG1655) | + | PlsrA-<i>gfp</i>-PlsrR-<i>lsrR</i> on pSB3K3(MG1655) |
[Method] | [Method] | ||
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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. | 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. | ||
| − | [[Image: | + | [[Image:Our_new_lsrA_promoter_activity%28BBa_K649105%29.png|thumb|center|500px|After four hours from OD590 reaching 0.15, we measured OD.<br>This work is done by Hiroki Yoshise.]] |
| − | + | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 14:46, 28 October 2011
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how you used this part and how it worked out.
Applications of BBa_K649105
We characterized BBa_K649105 in E.coli MG1655.
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.
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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User Reviews
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