Part:BBa_R0061:Experience
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UNIQ8d2ee9cad02448d5-partinfo-00000000-QINU
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iGEM Tokyo_Tech 2010
In order to characterize R0061, Plux repression promoter, we constructed K395101 combining R0061 and K121013, which is a promoter-less gfp reporter (rbs-gfp-ter-ter) on pSB6A1 and used a fusion of PlacIq (I14032) to gfp (K121013) as a positive control and used promoterless gfp (K121013) as a negative control.
Overnight cultures of reporter strains grown at 37 °C containing appropriated antibiotics were diluted at least 1:100 and incubated at 37 °C as fresh cultures. After their OD590 reached 0.6, added 3OC6HSL. After 3 hours of induction, fluorescence intensity was measured with flow cytometry.
After 3 hours of induction by 3OC6HSL, the expression of GFP with 3OC6HSL dropped to 1/3 comparing with the expression without 3OC6HSL.
We confirmed that this promoter works correctly.
UNIQ8d2ee9cad02448d5-partinfo-00000002-QINU
iGEM Chiba 2010
we also cheracterized about R0061.