Part:BBa_K649001: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_K649001
Fluorescence intensity of BBa_K649001 was increased by 3O-C12-HSL induction.
Generally, in the presence of 3O-C12-HSL lasI promoter is activated and the transcription level of downstream gene increases. To characterize this part, we used Ptrc-lasR on pBR as regulator part.
Method ①Overnight cultures of sample strain grown at 37 °C in LB medium containing carbenicillin and kanamycin were diluted 1:100 in the medium, and then they were incubated at 37 °C as fresh cultures. ②After their OD600 reached 0.2, we added 3 µL of 500 µM 3O-C12-HSL (3O-C12-HSL+) or 3µL of DMSO (3O-C12-HSL-) into the fresh cultures. ③After 3-hour incubation at 37 °C, 0.25 mL of each culture was harvested by centrifugalization and suspended by adding 1 mL of PBS (phosphate-buffered saline). ④We dispensed 500 µL of each suspension into a disposable tube through a cell strainer, and its fluorescence intensity was measured with a flow cytometer of Becton, Dickinson and Company.
User Reviews
UNIQa6a456ac35258cc5-partinfo-00000000-QINU UNIQa6a456ac35258cc5-partinfo-00000001-QINU