Genome location and transcriptional analysis of the h2cR sRNA in B. cenocepacia J2315.

(A) Genomic location of h2cR, showing in the upper part the 5′-UTR of thehfq2 gene, highlighting the direction of transcription of this gene (depicted by an open arrow and showing the ATG codon), the region where h2cR is inscribed (dark grey box), and the 5′-UTR region of hfq2 lacking h2cR complementarity (light grey box). A solid arrow in the lower part represents the h2cR transcript. The oligonucleotide used for 5′-RACE (5′R) experiments with h2cR is also shown. (B) h2cR RNA sequence and predicted secondary structure, showing the predicted folding energy. (C) Northern blot analysis of h2cR transcription in B. cenocepacia J2315 cells harvested from cultures grown for the indicated time. (D) Photograph of a 7.5% PAGE gel after electrophoresis of the PCR products obtained from 5′-RACE experiments of the h2cR sRNA, using the specific oligonucleotide 5′R, and 1 µg of total RNA obtained from B. cenocepacia J2315 cells in the exponential (2 h), late-exponential (8 h) of stationary (24 h) phases of growth.