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Fig. 3 | Microbial Cell Factories

Fig. 3

From: The cssR gene of Corynebacterium glutamicum plays a negative regulatory role in stress responses

Fig. 3

Negative regulation of ncgl1577 by CssR. a, e β-galactosidase analyses of the ncgl1577 promoter activities by using the transcriptional Pncgl1577::lacZY chromosomal fusion reporter expressed in WT(pXMJ19), ΔcssR(pXMJ19), and ΔcssR(pXMJ19-cssR) strains in the presence of GEN or CdCl2 conditions. b, f Quantitative RT-PCR analyses of ncgl1577 expression in WT(pXMJ19), ΔcssR(pXMJ19), and ΔcssR(pXMJ19-cssR) strains under GEN or CdCl2 conditions. The mRNA levels were presented relative to the value obtained from WT(pXMJ19) cells without stress treatment. Relative transcript levels of WT(pXMJ19) strains without stress treatment were set at a value of 1.0. c, g The protein levels of NCgl1577 in WT and ΔcssR in the presence or absence of GEN or CdCl2. Lysates from stationary phase bacteria exposed to GEN or CdCl2 for 2 h were resolved by SDS-PAGE, and NCgl1577 was detected by immunoblotting using specific anti-NCgl1577 antibody. For the pellet fraction, α-RNAP was used as a loading control. Similar results were obtained in three independent experiments, and data shown were from one representative experiment done in triplicate. d, h Relative quantified data for protein levels by Image Lab. Quantified protein expression of western blots in c and g. Densities of proteins were all justified with α-RNAP. Relative density ratios of WT without stress were set at a value of 1.0. Data shown were the averages of three independent experiments, and error bars indicated the SDs from three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001

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