Premium
Cloning and Characterization of Manganese Superoxide Dismutase Gene from Vibrio parahaemolyticus and Application to Preliminary Identification of Vibrio Strains
Author(s) -
Shyu YuChiau,
Lin FuPang
Publication year - 1999
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/713803521
Subject(s) - vibrio parahaemolyticus , escherichia coli , superoxide dismutase , recombinant dna , microbiology and biotechnology , open reading frame , molecular cloning , biology , biochemistry , gene , chemistry , enzyme , peptide sequence , bacteria , genetics
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 °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.