An Ochrobactrum anthropi gene conferring paraquat resistance to the heterologous host Escherichia coli

Abstract

A new gene, pqrA, conferring paraquat resistance to the heterologous host Escherichia coli, from a chromosomal DNA library of Ochrobactrum anthropi JW2, was cloned and analyzed. Cells of E. coli transformed with a plasmid carrying the pqrA gene showed elevated resistance to paraquat, but not to hydrogen peroxide. The predicted amino acid sequence of the PqrA polypeptide showed 71% identity with mll7495 hypothetical membrane protein in Mesorhizobium loti, 49% identity with PA2269 protein in Pseudomonas aeruginosa, and significant identity with other previously reported drug transport proteins. The hydropathy pattern of the PqrA polypeptide showed a significant homology to those of 12-transmembrane-segment (TMS) family export proteins. Immunoblot analysis demonstrated that the PqrA protein found in the membrane protein fraction of O. anthropi JW2 has a molecular mass of 42 kDa. These results suggest that the PqrA protein is a membrane protein that plays an important role in protecting cells against paraquat toxicity.