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

Revision as of 09:30, 2 October 2011

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

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.For this purpose, we constructed BBa_K649105(PlsrA-gfp-PlsrR-lsrR) and compared fluorescence intensity levels of BBa_K649104(PlsrA-RBS-gfp) and BBa_K649105.As a consequence, Median fluorescence intensity(MFI) of BBa_K649104(PlsrA-RBS-gfp) is 3-fold higher than that of BBa_K649105(PlsrA-gfp-PlsrR-lsrR). This result shows that LsrR represses lsrA promoter and BBa_K649105 works.

[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

UNIQeca3d1d98ae95805-partinfo-00000000-QINU UNIQeca3d1d98ae95805-partinfo-00000001-QINU