[Cloning and expression of superoxide dismutase gene from Deinococcus radiodurans in E. coli]

Sichuan Da Xue Xue Bao Yi Xue Ban. 2005 Mar;36(2):200-3.
[Article in Chinese]

Abstract

Objective: To construct expressing recombinant of Mn-SOD of Deinococcus radiodurans and express the target protein in E. coli BL21(DE3).

Methods: SOD gene was amplified by PCR from genomic DNA of Deinococcus radiodurans and inserted into expression plasmid pET-30a(+) to create the recombinant pET-SOD. After being analyzed by the restriction endonuclease, the plasmid was transformed into E. coli BL21(DE3), and the recombinant protein was expressed after induction by the isopropyl-beta-D-thiogalactopyranoside (IPTG) and was analyzed with SDS-PAGE.

Results: The recombinant plasmid pET-SOD was obtained, and the recombinant protein was highly expressed in E. coli BL21(DE3). The activity of recombinant superoxide dismutase was 51,800 U per gram of wet bacteria.

Conclusion: This study has provided a foundation for further studies and applications of the recombinant Mn-SOD.

MeSH terms

  • Cloning, Molecular
  • Deinococcus / enzymology
  • Deinococcus / genetics*
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Isopropyl Thiogalactoside
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Superoxide Dismutase / biosynthesis*
  • Superoxide Dismutase / genetics*

Substances

  • Recombinant Proteins
  • Isopropyl Thiogalactoside
  • Superoxide Dismutase