Identification, cloning and expression of sodC from an alkaline phosphatase gene fusion library of Mycobacterium avium subspecies paratuberculosis

Microbios. 2001:106 Suppl 1:7-19.

Abstract

The phoA gene technology was used to investigate secreted proteins of the intracellular pathogen Mycobacteriumn avium subspecies paratuberculosis. This led to the identification of sodC, a gene which codes for a copper and zinc cofactored superoxide dismutase (Cu,ZnSOD) which has been implicated as a virulence factor for some pathogens. The predicted protein possessed a 76% identity with Cu,ZnSOD of Mycobacterium tuberculosis. To characterize Cu,ZnSOD from M. avium subspecies paratuberculosis, the gene was cloned and overexpressed in Escherichia coli. The renatured, affinity-purified recombinant protein possessed enzymatic activity that was inhibited by the presence of cyanide, which is characteristic of a Cu,ZnSOD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / metabolism
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli Proteins*
  • Gene Library*
  • Molecular Sequence Data
  • Mycobacterium avium subsp. paratuberculosis / enzymology*
  • Mycobacterium avium subsp. paratuberculosis / genetics
  • Paratuberculosis / microbiology
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sequence Analysis, DNA
  • Superoxide Dismutase / chemistry
  • Superoxide Dismutase / genetics*
  • Superoxide Dismutase / metabolism*

Substances

  • Escherichia coli Proteins
  • Recombinant Fusion Proteins
  • Superoxide Dismutase
  • sodC protein, E coli
  • Alkaline Phosphatase

Associated data

  • GENBANK/AF326234