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Expression, purification, and crystallization of restriction endonuclease Pvu II with DNA containing its recognition site
Author(s) -
Balendiran K.,
Bonventre Joseph,
Knott Roger,
Jack William,
Benner Jack,
Schildkraut Ira,
Anderson John E.
Publication year - 1994
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.340190110
Subject(s) - restriction enzyme , microbiology and biotechnology , orthorhombic crystal system , endonuclease , oligonucleotide , dna , biology , duplex (building) , enzyme , crystallization , chemistry , crystallography , biochemistry , crystal structure , organic chemistry
We have overexpressed the type II restriction endonuclease Pvu II (R. Pvu II) in E. coli , prepared large amounts of the homogeneous enzyme, and crystallized it with an oligonucleotide carrying a Pvu II recognition site. The cocrystals are orthorhombic space group P 2 1 2 1 2 1 with cell constants a = 95.8 Å, b = 86.3 Å, c = 48.5 Å, and diffract X‐rays to at least 2.7 Å. There is a complex of two protein subunits and one oligonucleotide duplex in the asymmetric unit. © 1994 Wiley‐Liss, Inc.