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Fig. 4 | BMC Microbiology

Fig. 4

From: 6S RNA plays a role in recovery from nitrogen depletion in Synechocystis sp. PCC 6803

Fig. 4

Analysis of 6S RNA transcript levels during nitrogen starvation and recovery in Synechocystis 6803 WT (a) and physiological characterization of the Synechocystis 6803 mutant strain overexpressing 6S RNA (6S(+)) (b and c). a Total RNA was isolated from cell cultures grown at standard growth conditions (t = Std), under nitrogen depleted conditions (t = 3 h -N, t = 6 h -N, t = 24 h -N, t = 189 h -N) and under nitrogen replete conditions (t = 4 h + N, t = 22 h + N, t = 48 h + N, t = 144 h + N) and samples with 5 μg RNA were analyzed by Northern blot hybridization. Autoradiograms are illustrated for WT I. 6S RNA and 5S rRNA signals of two biological replicates (WT I, WT II) were quantitated and 5S rRNA signal was used as an internal reference. The mean values are illustrated in the chart. Values of t = Std were set to 100 and samples of all other time points were calculated, respectively. b Northern Blot analysis to confirm overexpression of 6S RNA in 6S(+) mutant. 2 μg of total RNA was separated by electrophoresis on 1.3% agarose-formaldehyde gel. The amount of 6S RNA and 5S rRNA was detected by using radioactive [32P]-labeled oligonucleotides. c Physiological characterization of the 6S(+) strain. Absorbance spectra of the Synechocystis 6803 wild-type and the 6S(+) strain are illustrated for the time points t = 0 h + N, t = 4 h + N, t = 22 h + N and t = 48 h + N. The spectra were normalized at 750 nm. C: carotenoids; PC: phycocyanine; Chl: chlorophyll a

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