Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase

Gene. 1988 Jul 15;67(1):31-40. doi: 10.1016/0378-1119(88)90005-4.


Plasmid expression vectors have been constructed that direct the synthesis of foreign polypeptides in Escherichia coli as fusions with the C terminus of Sj26, a 26-kDa glutathione S-transferase (GST; EC encoded by the parasitic helminth Schistosoma japonicum. In the majority of cases, fusion proteins are soluble in aqueous solutions and can be purified from crude bacterial lysates under non-denaturing conditions by affinity chromatography on immobilised glutathione. Using batch wash procedures several fusion proteins can be purified in parallel in under 2 h with yields of up to 15 micrograms protein/ml of culture. The vectors have been engineered so that the GST carrier can be cleaved from fusion proteins by digestion with site-specific proteases such as thrombin or blood coagulation factor Xa, following which, the carrier and any uncleaved fusion protein can be removed by absorption on glutathione-agarose. This system has been used successfully for the expression and purification of more than 30 different eukaryotic polypeptides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Escherichia coli / genetics*
  • Genes*
  • Genetic Vectors
  • Glutathione Transferase / genetics
  • Glutathione Transferase / isolation & purification*
  • Molecular Sequence Data
  • Plasmids
  • Recombinant Fusion Proteins / isolation & purification*
  • Recombinant Proteins / isolation & purification*
  • Schistosoma japonicum / enzymology
  • Schistosoma japonicum / genetics


  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Glutathione Transferase

Associated data

  • GENBANK/M21676