Molecular cloning and characterization of superoxide dismutase gene from Vibrio metschnikovii

• Main Authors: ma hsiao fen, 馬小芬
• Other Authors: lin fu pang
• Format: Others
• Language: zh-TW
• Published: 1999
• Online Access: http://ndltd.ncl.edu.tw/handle/49802720259568268704

碩士 === 國立海洋大學 === 水產生物技術研究所 === 87 === The superoxide dismutase gene from Vibrio metschnikovii was cloned successfully by PCR techniques using the degenerate Mn-SOD No and Mn-SOD Co primers, and the random primer gene walking PCR using V. metschnikovii sod gene specific primers and random hexamer. The full-length sod gene was composed of 588 base pair and encoded 196 amino acid residues. Its amino acid sequence showed high homology to Photobacterium leiognathi, E. coli Fe-sod gene (85.5%, 76.2%, respectively). V. metschnikovii sod gene was cloned into the expression vector pET20b(+) and expressed in E. coli BL21(DE3)pLysS. Biochemical characteristics of the purified recombinant Vme-SOD have confirmed it encoded a Fe- or Mn-SOD. The specificity of V. metschnikovii sod gene probe was analyzed by cross-species PCR in order to provide the preliminary identification of Vibrio strains.