Figure 8.

Accumulation of Hfq and GcvB throughout growth. A, accumulation of Hfq. Overnight cultures of the WT strain carrying chromosomal 3×FLAG-tagged hfq was diluted into 100 ml of fresh YT medium, and cells were removed at the indicated time points. Ten μg of total protein was resolved by 15% SDS-PAGE and subjected to immunoblotting using antibody directly against the FLAG tag. Coomassie Blue staining was used as the loading control. B, accumulation of GcvB RNA. Fifteen μg of total RNA was separated by 6% urea-PAGE and subjected to Northern blot analysis using probes specific for GcvB and 5S RNA. A representative experiment carried out twice with essentially identical results is shown. The relative amount of Hfq or GcvB is indicated with average ± S.D. from the two biological replicates by comparing to the sample at A₆₀₀ = 0.2, which was set at 1.0.