Cloning, Expression and Gene Probe Speificity of the Managanese Superoxide Dismutase Gene from Vibrio Mimicus

• Main Author: 金志宇
• Other Authors: 林富邦
• Format: Others
• Language: zh-TW
• Published: 1998
• Online Access: http://ndltd.ncl.edu.tw/handle/56366724762477773887

碩士 === 國立海洋大學 === 水產生物技術研究所 === 86 === 　　Superoxide dismutases (SODs) are metalloenzymes that detoxify superoxide radicals present in various living cells. A gene encoding manganese superoxide dismutase (Mn-SOD) was cloned successfully from Vibrio mimicus by both traditional and random primer gene walking PCR which using the combination of degenerate and gene specific primers. The full-length Mn-SOD gene was composed of 606 base pair and encoded a 202 amino acid open reading frame.The amino acid sequence exhibits a 56.9% indentity to the Mn-SOD from Bacillus caldotenax and shows an overall homology to sequences of other known Mn-SODs. The V. mimicus Mn-SOD gene was introduced into the pET20b(+) expression vector and then transformed into the E. coli BL21 (DE3) pLysS expression host for the heterologous gens expression. The specificity of V. mimicus Mn-SOD gene probe was analyzed by PCR among several Vibrio spp. tested in order to provide the preliminary information for the purpose of Vibrio strain identification.