PUBLICATION
Characterization of fish Cu/Zn-superoxide dismutase and its protection from oxidative stress
- Authors
- Ken, C.F., Lin, C.T., Shaw, J.F., and Wu, J.L.
- ID
- ZDB-PUB-030728-17
- Date
- 2003
- Source
- Marine biotechnology (New York, N.Y.) 5(2): 167-173 (Journal)
- Registered Authors
- Shaw, Jei-Fu, Wu, Wen-Jun
- Keywords
- none
- MeSH Terms
-
- Animals
- Cloning, Molecular
- DNA, Complementary/genetics
- Escherichia coli/genetics
- Herbicides/pharmacology
- Larva/drug effects
- Larva/metabolism
- Oxidative Stress/drug effects
- Oxidative Stress/physiology*
- Paraquat/pharmacology
- Superoxide Dismutase/genetics
- Superoxide Dismutase/isolation & purification
- Superoxide Dismutase/metabolism*
- Zebrafish/genetics
- Zebrafish/metabolism*
- Zebrafish Proteins/genetics
- Zebrafish Proteins/isolation & purification
- Zebrafish Proteins/metabolism
- PubMed
- 12876653 Full text @ Mar. Biotechnol.
Citation
Ken, C.F., Lin, C.T., Shaw, J.F., and Wu, J.L. (2003) Characterization of fish Cu/Zn-superoxide dismutase and its protection from oxidative stress. Marine biotechnology (New York, N.Y.). 5(2):167-173.
Abstract
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 degrees 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 micro g/ml SOD enzyme. The results were significant.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping