Expression, purification, and crystallization of restriction endonuclease PvuII with DNA containing its recognition site

Proteins. 1994 May;19(1):77-9. doi: 10.1002/prot.340190110.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Sequence
  • Binding Sites
  • Crystallization
  • Crystallography, X-Ray*
  • Deoxyribonucleases, Type II Site-Specific / chemistry*
  • Deoxyribonucleases, Type II Site-Specific / isolation & purification
  • Deoxyribonucleases, Type II Site-Specific / metabolism
  • Escherichia coli
  • Gene Expression
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides / metabolism*
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism

Substances

  • Oligodeoxyribonucleotides
  • Recombinant Fusion Proteins
  • CAGCTG-specific type II deoxyribonucleases
  • Deoxyribonucleases, Type II Site-Specific