Microtiter plate-based assay of MicC-mediated fusion repression

Copyright information:

Taken from "Translational control and target recognition by small RNAs "

Nucleic Acids Research 2007;35(3):1018-1037.

Published online 30 Jan 2007

PMCID:PMC1807950.

© 2007 The Author(s).

() strains carrying the fusion in combination with one of the 10 sRNA expression plasmids or the pJV300 control plasmid (specified by the color code) were inoculated in 150 μl LB medium in a 96-well microtiter plate, overlaid with mineral oil and grown with agitation at 37°C in a Victor3 plate reader for 16 h. Cell density and fluorescence were determined in 15 min intervals. Plotting of fluorescence values over growth (OD) shows specific repression of the fusion by the MicC expression plasmid. Cell density is given as OD values of 600 nm light absorption in the microtiter plate well (0.2 OD units corresponds to ∼1 OD standard units). Fluorescence values were corrected for the autofluorescence of an strain carrying control plasmids pXG-0 and pJV300. () Factor of fusion regulation by sRNAs (relative to the control plasmid pJV300, calculated as in ) at five selected growth stages (OD of 0.1, 0.15, 0.2, 0.25 and 0.3). Black bars indicate repression, whereas gray bars activation.