Article Sidebar
Published:
Jun 26, 2013DOI:
10.4314/tjpr.v12i3.9Keywords:
Article Details
Submission of a manuscript to this journal is a representation that the manuscript has not been published previously and is not under consideration for publication elsewhere.
All authors named in each manuscript would be required to sign a form (to be supplied by the Editor) so that they may retain their copyright in the article but to assign to us (the Publishers) and its licensees in perpetuity, in all forms, formats and media (whether known or created in the future) to (i) publish, reproduce, distribute, display and store the contribution, (ii) translate the contribution into other languages, create adaptations, reprints, include within collections and create summaries, extracts and/or abstracts of the contribution, (iii) create any other derivative works(s) based on the contribution, (iv) to exploit all subsidiary rights in the contribution, (v) the inclusion of electronic links from the contribution to third party material where-ever it may be located, and (vi) license any thrid party to do any or all of the above.
Main Article Content
Expression, Purification, Characterization and In Vitro Activity of Recombinant Mouse Cu/Zn-Binding Superoxide Dismutase (mSOD1)
Abstract
Purpose: To express, purify and characterize recombinant mouse Cu/Zn-binding superoxide dismutase (mSOD1), and investigate its activity in vitro.
Methods: The protein, mSOD1, was expressed after induction with isopropyl-beta-Dthiogalactopyranoside (IPTG). The target protein was purified by Ni-NTA affinity chromatography. The identity of the recombinant protein was confirmed by Western-blot and peptide mass fingerprinting
(PMF) analysis. Protein activity in vitro was investigated by SOD activity assay kit and DNA damage protective assay.
Results: mSOD1 protein was expressed with a final yield of about 60 mg of pure protein per liter of culture medium. After purification, the target protein was > 95 %. The identity of the recombinant protein was confirmed. SOD activity assay showed that the highest activity of the mSOD1 was 3789.0 ± 80.5 U/mg. The present work showed that mSOD1 was effective in protecting DNA from oxidative damage.
Conclusion: High purity recombinant mSOD1 was obtained and characterized, and had high activity in vitro. The study indicates that the mSOD1 may serve as a potential therapeutic agent for those diseases
caused by oxidative stress.
Keywords: Cu/Zn-binding Superoxide dismutase, Expression, Purification, Metal ions, DNA damage
