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 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.
Foster
Kimberly A
KA
Microbiotix, Inc., One Innovation Drive, Worcester, MA 01605, USA. kfoster@microbiotix.com
Barnes
Marjorie H
MH
Stephenson
Ryan O
RO
Butler
Michelle M
MM
Skow
Donna J
DJ
LaMarr
William A
WA
Brown
Neal C
NC
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D004258:DNA Polymerase III / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D013293:Enterococcus faecalis / Q000201:enzymology* / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli; D015870:Gene Expression; D005798:Genes, Bacterial / Q000235:genetics*; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D006160:Guanosine Triphosphate; C074904:DNA polymerase III, alpha subunit; C446929:PolC protein, bacteria; D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase