PURIFICATION, CRYSTALLIZATION AND PRELIMINARY-X-RAY CRYSTALLOGRAPHIC ANALYSIS OF PYROCOCCUS-FURIOSUS DNA-POLYMERASE

DNA polymerase gene from the hyperthermophilic Archaeon Pyrococcus fur iosus has been cloned and the protein overexpressed in Escherichia coi l to produce an active enzyme. The purified protein was crystallized f rom 0.08 M ammonium sulfate, 0.05 M Na-cacodylate, pH 6.5, 0.15%(v/v) NP40, 0.05%(v/v) Tween 20 and 4.5% (w/v) polyethylene glycol 6000 by t he vapour-diffusion method. The orthorhombic crystals had unit-cell di mensions of a = 92.5, b = 125.4, c = 192.1 Angstrom: alpha = beta = ga mma = 90 degrees. The crystals diffracted beyond 4 Angstrom on a 1.08 Angstrom synchrotron radiation source.