Overexpression, purification and crystallization of BamHI endonuclease

Nucleic Acids Res. 1991 Apr 25;19(8):1825-9. doi: 10.1093/nar/19.8.1825.

Abstract

The type II restriction endonuclease BamHI has been expressed in E. coli, producing 100-fold more enzyme than the wild type Bacillus amyloliquefaciens H strain. This high yield has facilitated purification to homogeneity of large amounts of the enzyme, along with its crystallization in a form which diffracts to at least 1.9 A in X-ray analysis.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacillus / enzymology
  • Cloning, Molecular
  • Crystallization
  • Deoxyribonuclease BamHI / biosynthesis*
  • Deoxyribonuclease BamHI / chemistry
  • Deoxyribonuclease BamHI / isolation & purification
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics*
  • Plasmids
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • X-Ray Diffraction

Substances

  • Recombinant Proteins
  • Deoxyribonuclease BamHI