1. Expression, purification, and crystallization of restriction endonucleasePvuII with DNA containing its recognition site
- Author
-
Jack S. Benner, Ira Schildkraut, William E. Jack, John E. Anderson, Roger Knott, K. Balendiran, and Joseph V. Bonventre
- Subjects
Stereochemistry ,Recombinant Fusion Proteins ,Protein subunit ,Molecular Sequence Data ,Gene Expression ,Protein-DNA complex ,Biology ,Crystallography, X-Ray ,Biochemistry ,chemistry.chemical_compound ,Structural Biology ,Gene expression ,Escherichia coli ,Deoxyribonucleases, Type II Site-Specific ,Molecular Biology ,chemistry.chemical_classification ,Binding Sites ,Base Sequence ,Oligonucleotide ,Molecular biology ,Restriction enzyme ,Enzyme ,Oligodeoxyribonucleotides ,chemistry ,Orthorhombic crystal system ,Crystallization ,DNA - Abstract
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 A, b = 86.3 A, c = 48.5 A, and diffract X-rays to at least 2.7 A. There is a complex of two protein subunits and one oligonucleotide duplex in the asymmetric unit.
- Published
- 1994
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