DNA polymerase III of Enterococcus faecalis: expression and characterization of recombinant enzymes encoded by the polC and dnaE genes

Protein Expr Purif. 2003 Jan;27(1):90-7. doi: 10.1016/s1046-5928(02)00577-6.


Enterococcus faecalis (Ef) dnaE and polC, the respective genes encoding the DNA replication-specific DNA polymerase III E and DNA polymerase III C, were cloned and engineered for expression in Escherichia coli as hexahistidine (his6)-tagged recombinant proteins. Each gene expressed a catalytically active DNA polymerase of the expected molecular weight. The recombinant polymerases were purified and each was characterized with respect to catalytic properties, inhibitor sensitivity, and recognition by specific antibody raised against the corresponding DNA polymerase III of the model Gram-positive (Gr(+)) organism, Bacillus subtilis (Bs). In conclusion, the properties of each Enterococcus polymerase enzymes were similar to those of the respective B. subtilis enzymes.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins*
  • Base Sequence
  • Cloning, Molecular
  • DNA Polymerase III / chemistry
  • DNA Polymerase III / genetics*
  • DNA Polymerase III / isolation & purification
  • DNA Polymerase III / metabolism*
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / genetics*
  • DNA-Directed DNA Polymerase / isolation & purification
  • DNA-Directed DNA Polymerase / metabolism
  • Enterococcus faecalis / enzymology*
  • Enterococcus faecalis / genetics
  • Enzyme Inhibitors / pharmacology
  • Escherichia coli
  • Gene Expression
  • Genes, Bacterial / genetics*
  • Guanosine Triphosphate / analogs & derivatives
  • Guanosine Triphosphate / pharmacology
  • Molecular Sequence Data
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics*
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism*
  • Sequence Analysis, DNA


  • Bacterial Proteins
  • Enzyme Inhibitors
  • Recombinant Proteins
  • Guanosine Triphosphate
  • DNA polymerase III, alpha subunit
  • PolC protein, bacteria
  • DNA Polymerase III
  • DNA-Directed DNA Polymerase