Difference between revisions of "Part:BBa K649105:Experience"

Revision as of 11:55, 2 October 2011

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Applications of BBa_K649105

After four hours from OD590 reaching 0.15, we measured OD.
This work is done by Hiroki Yoshise.

Median fluorescence intensity(MFI) is decreased by LsrR repression.
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 PlsrA-gfp.By the LsrR repression, the fluorescence intensity decreased 3-fold. (Fig.6). This result shows that LsrR successfully repressed PlsrA. The working parts 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]

pSB6A1 Ptet-GFP RBS1-12(JM2.300)(positive control)

pSB6A1 ⊿P-GFP(JM2.300)(negative control)

pSB3K3 PlsrA-GFP(MG1655)

pSB3K3 PlsrA-GFP-PlsrR-lsrR(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.

User Reviews

UNIQ1d3c204480bf2661-partinfo-00000000-QINU UNIQ1d3c204480bf2661-partinfo-00000001-QINU