Portal de Periódicos da CAPES

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

2002; Elsevier BV; Volume: 27; Issue: 1 Linguagem: Inglês

10.1016/s1046-5928(02)00577-6

1096-0279

Kimberly A. Foster, Marjorie H. Barnes, Ryan O. Stephenson, Michelle M. Butler, Donna Skow, William A. LaMarr, Neal C. Brown,

Bacteriophages and microbial interactions

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.