The rrnA promoter as a tool for the improved expression of heterologous genes in cyanobacteria

The regulatory sequence of ribosomal RNA A (rrnA) operon from Synechococcus PCC7942 was characterized using green fluorescent protein gene (gfp) as a reporter. The PR promoter (nt. −83 to +2) including upstream promoter element and P1 promoter of rrnA exhibited GFP fluorescence intensity about 30-fold higher than full length sequence (nt. −147 to +79). The effects of PR promoter arranged in tandem with consensus-σ70 promoter (PS) of Escherichia coli on the expression of gfp and opd gene encoding organophosphorus hydrolase (OPH) in Synechococcus were investigated. The PS-PR tandem promoter was superior to all of the other promoters; its GFP fluorescence intensity was a 1.8-fold increase when compared to PR-PR tandem promoter, a 2.5-fold, 9.5-fold and a 15-fold increase compared to PR, PS and promoter of tRNApro, respectively. The GFP from PS-PR tandem promoter accounted for about 12% of its total extracted proteins. OPH activity of Synechococcus harboring opd gene under the control of PS-PR tandem promoter was 738 ± 128 units/OD730. We demonstrated that the tandem promoters remarkably enhanced the GFP and OPH production which were detected on SDS-PAGE stained with Coomassie blue. The promoter system in this study could be generally applied to production of valuable organic products from cyanobacteria.