Fluorescence and expression characteristics of representative fusions

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).

See Table 1 and Supplementary Table S3 for details of fusion plasmids. () strains carrying control plasmids (no = pXG-0; = pXG-1) or target fusion plasmids (as indicated) were grown on LB agar. The left image was obtained in the visible light mode and shows the colony morphology of these strains. The right image shows the same plate in the fluorescence mode. GFP fluorescence was excitated at 460 nm, and light emission was recorded using a 510 nm-filter. () Fluorescence of cells carrying the indicated fusions at different cell densities. Bacteria were grown aerobically in liquid culture in triplicates and aliquots were measured at the indicated cell density (OD). Fluorescence values are given in arbitrary units and were corrected for the basal fluorescence of an strain harboring plasmid pXG-0 (∼40 000 U). The left panel shows a set of low fluorescence fusions; the right panel shows fusions that yielded high fluorescence and includes plasmid pXG-1 expressing full-length GFP. () Detection of GFP fusion proteins and fusion mRNAs. Samples were taken from liquid cultures of strains carrying the control plasmid pXG-1 or the indicated fusion plasmids at OD of 0.5 and 1, and were subjected to western blot analysis with monoclonal α-GFP antibodies (upper panel) and to northern analysis (middle panel). The same northern blot was probed for 5S rRNA as a loading control (lower panel).