Single-step purification of bacterially expressed polypeptides containing an oligo-histidine domain

Gene. 1992 Feb 1;111(1):99-104. doi: 10.1016/0378-1119(92)90608-r.


Plasmid expression vectors have been constructed that direct the synthesis in Escherichia coli of fusion proteins containing a stretch of six histidine residues at either the N or C terminus. This oligo-histidine domain allows the single-step purification of the fusion proteins, under nondenaturing conditions, by immobilized metal affinity chromatography on Ni2+ bound to iminodiacetic acid-agarose. Several eukaryotic transcription factors (e.g., the upstream stimulatory factor for the adenovirus major late promoter) have been successfully purified, in an active state, by this method.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification*
  • Base Sequence
  • Chromatography, Affinity
  • Cloning, Molecular
  • DNA, Bacterial
  • Escherichia coli / genetics
  • Genetic Vectors*
  • Histidine / genetics*
  • Molecular Sequence Data
  • Plasmids*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification


  • Bacterial Proteins
  • DNA, Bacterial
  • Recombinant Fusion Proteins
  • Histidine