The rrnA promoter as a tool for the improved expression of heterologous genes in cyanobacteria
| Autor: | Sirirat Fa-aroonsawat, Wipa Chungjatupornchai |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Genetics
Microbial Operon Green Fluorescent Proteins Gene Expression Sigma Factor Biology medicine.disease_cause Microbiology Green fluorescent protein Genes Reporter Escherichia coli medicine Promoter Regions Genetic Molecular Biology Gene Synechococcus Aryldialkylphosphatase Promoter DNA-Directed RNA Polymerases Ribosomal RNA Molecular biology Recombinant Proteins Artificial Gene Fusion Regulatory sequence RRNA Operon |
| Zdroj: | Microbiological Research. 169:361-368 |
| ISSN: | 0944-5013 |
| DOI: | 10.1016/j.micres.2013.09.010 |
| Popis: | 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-σ(70) 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 tRNA(pro), 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. |
| Databáze: | OpenAIRE |
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