Cloning, Expression and Characterization of PprI gene in Escherichia coli

Abstract

Background and Objectives: PprI is one of newly gene identified in Deinococcus radiodurans and plays a critical role in DNA repair and protection against ultraviolet radiation stress. The aim of this study was to clone, express and characterize PprI gene in Escherichia coli.Materials and Methods: The PprI gene of D. radiodurans was constructed in pGEM-B1 vector and the cloned gene was subcloned into pET21a expression vector. The pET21a-PprI was expressed in E. coli (Origami strain). Expression of recombinant PprI was confirmed by SDS-PAGE gel electrophoresis and western blotting. In addition, the UV-C radiation resistance of recombinant E. coli was determined.Results: The chimeric pET21a plasmid containing the C -terminal fusion of His-tag with PprI gene was successfully constructed and the medium condition for the expression was optimized. In addition, transformed E. coli had higher resistance to radiation than the original strain. Conclusion: The results of this study indicated that PprI protein could be a potential candidate to be considered as a radioresistant protein. However, the UV-C radioresistance potency of recombinant PprI should be analyzed in prokaryotic and eukaryotic systems

Keywords

• PprI gene
• Radiation resistant
• Deinococcus radiodurans