Superoxide dismutase cloning and expression in microorganisms

Abstract

Abstract The invention provides a DNA construct comprising a functional vector for expression in E. coli comprising in downstream order of transcription: (1) a promoter, (2) a ribosomal binding site, (3) an initiation codon, (4) a human cytoplasmic Cu/Zn superoxide dismutase structural gene, or alleles thereof, in reading frame with said initiation codon having a stop codon at the 3'-terminus of said gene and (5) a transcription terminator, wherein said DNA construct, upon expression in E. coli, expresses non-acetylated active human cytoplasmic Cu/Zn superoxide dismutase. The invention also provides a method for preparing a polypeptide comprising non-acetylated enzymatically active human cytoplasmic Cu/Zn superoxide dismutase that differs by no more than four amino acids from naturally-occurring human cytoplasmic Cu/Zn superoxide dismutase, said method comprising growing E. coli containing a DNA construct of the invention in a nutrient medium and isolating the resulting non-acetylated enzymatically active human cytoplasmic superoxide dismutase.

Inventors

• 👤 Robert A. Hallewell
• 👤 Guy T. Mullenbach

Bibliographic Details