EXPRESSION, PURIFICATION, AND CRYSTALLIZATION OF RESTRICTION-ENDONUCLEASE PVUII WITH DNA CONTAINING ITS RECOGNITION SITE

We have overexpressed the type II restriction endonuclease PvuII (R-PvuII) in E. coli, prepared large amounts of the homogeneous enzyme, and crystallized it with an oligonucleotide carrying a PvuII recognition site. The cocrystals are orthorhombic space group P2(1)2(1)2(1) with cell constants a = 95.8 Angstrom, b = 86.3 Angstrom, c = 48.5 Angstrom, and diffract X-rays to at least 2.7 hi. There is a complex of two protein subunits and one oligonucleotide duplex in the asymmetric unit. (C) 1994 Wiley-Liss, Inc.