title: Cloning and characterization of manganese superoxide dismutase gene from Vibrio parahaemolyticus and application to preliminary identification of Vibrio strains.
abstract: The sodA gene coding for manganese superoxide dismutase (Mn-SOD) from the marine microorganism Vibrio parahaemolyticus was cloned, sequenced, and overexpressed in Escherichia coli by use of the pET20b (+) expression vector. The full-length gene consisted of a 588-bp open reading frame and encoded a polypeptide of 196 amino acid residues, with a calculated molecular mass of 21,713 Da. The recombinant enzyme was efficiently purified from crude E. coli cell lysate by metal ion affinity chromatography. The recombinant VPMn-SOD resisted thermo-denaturation up to 60 degrees C and was insensitive to such inhibitors as EDTA, NaN3 and diethyldithiocarbamic acid. The specificity of V. parahaemolyticus Mn-SOD gene probe was analyzed by cross-species polymerase chain reaction to provide information for Vibrio strain identification.
authors.affiliation: Institute of Marine Biotechnology, National Taiwan Ocean University, R.O.C.
authors.forename: Y C
authors.initials: YC
authors.lastname: Shyu
authors.forename: F P
authors.initials: FP
authors.lastname: Lin
mesh_terms: D000595:Amino Acid Sequence; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D013482:Superoxide Dismutase / Q000235:genetics*; D014736:Vibrio parahaemolyticus / Q000235:genetics*
chemical_list: D013482:Superoxide Dismutase