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

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 a...

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Bibliographic Details
Published in:Protein expression and purification 2003-01, Vol.27 (1), p.90-97
Main Authors: Foster, Kimberly A, Barnes, Marjorie H, Stephenson, Ryan O, Butler, Michelle M, Skow, Donna J, LaMarr, William A, Brown, Neal C
Format: Article
Language:English
Subjects:
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
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Summary: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.
ISSN:1046-5928
DOI:10.1016/S1046-5928(02)00577-6