Part:BBa_K649105:Experience

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.
Amp　FSC：E01　SSC:450　FL1:730
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. 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(JM2.300)(positive control)

pSB6A1 promoterless 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

UNIQ6c66b929e7d7cd91-partinfo-00000000-QINU UNIQ6c66b929e7d7cd91-partinfo-00000001-QINU