Cloning and expression of Deinococcus radiodurans drRRA gene in E. coli Origami strain

Aim and Deinococcus radiodurans is one of the most radio resistant microorganisms able to survive in an extreme environment. Available data show that multiple DNA repair and antioxidant proteins are involved in radio resistance. DrRRA is a novel response regulator essential for the radio resistant of Deinococcus radiodurans. The aim of this study was cloning and expression of drRRA gene in E. coli system. The drRRA gene of Deinococcus radiodurans was synthesized in pGEM-B1 vector and the synthetic gene was subcloned to pET21a expression vector. Subcloning of the gene was verified by double digestion and sequencing. The cloned gene was expressed in E. coli (Origami strain). Expression of recombinant DrRRA (rDrRRA) was confirmed by polyacrylamide gel electrophoresis and Western blotting. The chimeric pET21a plasmid containing the C-terminal fusion of His-tag with DrRRA was successfully constructed. Recombinant DrRRA was intracellularly produced by transformed E. coli. The results of this study indicated that subcloning and expression of drRRA gene in E. coli is feasible. In addition, the expression level of protein was suitable and it should be analyzed in prokaryotic and eukaryotic systems.