Fig. 8

Analysis of fimA (XAC3241) mRNA stability. a The predicted secondary structure of the fimA 5′ untranslated region was obtained using MFold software [32]. The first codon, AUG, is shown in a red box. b Analysis of fimA mRNA abundance in the wild-type, ∆hrpB and complementation (∆hrpB-p53hrpB) strains was performed using RT-PCR. Analysis of the wild-type, ∆hrpB and complementation (∆hrpB-p53hrpB) strains that were grown in NB medium to OD 600 nm = 0.5 and then treated with 10 mg/mL ciprofloxacin before being harvested at several time points. fimA mRNA stability was then determined in the wild-type and ∆hrpB strains using RT-PCR. Total RNA was isolated, and 2 mg of RNA was used to synthesize the cDNA that was used for RT-PCR in 25 mL reactions. The reactions were subjected to PCR amplification for 22 cycles. Ten microliters of each reaction was resolved on a 1.5 % agarose gel. The stability of the fimA transcript was evaluated, and gyrA was used as the control to normalize the fimA amplification products