National Changhua University of Education Institutional Repository : Item 987654321/16770
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题名: Characterization of Fish Cu/Zn–Superoxide Dismutase and Its Protection from Oxidative Stress
作者: Ken, Chuian-Fu;Lin, Chi-Tsai;Shaw, Jei-Fu;Wu, Jen-Leih
贡献者: 生物學系
关键词: Cu/Zn superoxide dismutase;Overexpression;Zebrafish;Paraquat;Escherichia coli
日期: 2003-04
上传时间: 2013-06-05T08:48:10Z
出版者: Springer-Verlag
摘要: Copper/zinc superoxide dismutase was cloned from the zebrafish (Danio rerio). The full coding region of the zebrafish superoxide dismutase (ZSOD) complementary DNA was ligated with pET-20b(+) and successfully expressed in Escherichia coli strain AD494(DE3)pLysS. The active enzyme was purified by His tagging. The ZSOD yield was 6 mg from 0.2 L of E. coli culture, and the specific activity was 2000 U/mg as assayed using a RANSOD kit. The enzyme stability was characterized by reaction to temperature, pH, and detergent treatment. The results showed enzyme activity was still active after heat treatment at 70�C for 10 minutes, resistant to pH treatment from 2.3 to 12, and resistant to treatment with sodium dodecyl sulfate (SDS) under 4%. In addition, the recombinant ZSOD was used to protect fish from 100 ppm of paraquat-induced oxidative injury by soaking fish larva in 55 �g/ml SOD enzyme. The results were significant.
關聯: Marine Biotechnology, 5(2): 167-173
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