Open AccessExpression, purification and preliminary X‐ray analysis of proliferating cell nuclear antigen from the archaeon Thermococcus thioreducens
Author(s) -
ByrneSteele Miranda L.,
Ng Joseph D.
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910903036x
Subject(s) - proliferating cell nuclear antigen , biology , thermococcus , escherichia coli , crystallography , dna , dna polymerase , microbiology and biotechnology , antigen , chemistry , archaea , biochemistry , gene , genetics
Proliferating cell nuclear antigen (PCNA) is a DNA sliding clamp which confers processivity on replicative DNA polymerases. PCNA also acts as a sliding platform that enables the association of many DNA‐processing proteins with DNA in a non‐sequence‐specific manner. In this investigation, the PCNA from the hyperthermophilic archaeon Thermococcus thioreducens (TtPCNA) was cloned, overexpressed in Escherichia coli and purified to greater than 90% homogeneity. TtPCNA crystals were obtained by sitting‐drop vapor‐diffusion methods and the best ordered crystal diffracted to 1.86 Å resolution using synchrotron radiation. The crystals belonged to the hexagonal space group P 6 3 , with unit‐cell parameters a = b = 89.0, c = 62.8 Å. Crystals of TtPCNA proved to be amenable to complete X‐ray analysis and future structure determination.