FIGURE 4.

OpaR regulates the transcription of c-di-GMP metabolism-related genes in V. parahaemolyticus RIMD2210633. The negative and positive numbers indicate the nucleotide positions upstream and downstream of indicated genes, respectively. Bacterial cells were harvested after 6 h of incubation at 37°C on HI plates, and total RNAs were extracted using TRIzol Reagent. (A) qPCR assay was used to determine the relative mRNA levels for each target gene in ΔopaR and WT using a standard curve of 16S rRNA expression. (B) LacZ fusion. The regulatory DNA region of each target gene was cloned into the pHRP309 plasmid and then transferred into ΔopaR and WT to determine the promoter activity in cellular extracts.