Production of human β-actin and a mutant using a bacterial expression system with a cold shock vector

Protein Expr Purif. 2011 Jul;78(1):1-5. doi: 10.1016/j.pep.2010.09.007. Epub 2010 Sep 17.


Actin is the most abundant protein in the cytoplasm of most eukaryotic cells and is involved in a variety of cellular functions. It has been difficult to produce actin in bacterial expression systems in good yields. In this study, we developed a new simple method for the production of recombinant actin in Escherichia coli cells. Human β-actin was successfully expressed using a cold shock vector, pCold, in the bacterial expression system. The expressed β-actin (hexahistidine-tagged) was separated with a Ni-chelating resin, followed by a polymerization/depolymerization cycle or column chromatography with the Ni-chelating resin. The purified recombinant β-actin showed a normal polymerization ability compared with β-actin purified from human platelets. We produced a recombinant mutant actin with a Gly-168Arg mutation in the system and confirmed that it exhibited an impaired polymerization ability. The system developed in this study will provide a useful method for the production of actin isoforms and their mutants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / biosynthesis*
  • Actins / genetics
  • Actins / isolation & purification
  • Actins / metabolism
  • Biotechnology / methods
  • Blotting, Western
  • Chromatography, Affinity
  • Cold Shock Proteins and Peptides / genetics*
  • Escherichia coli / chemistry
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Genetic Vectors / genetics
  • Histidine / chemistry
  • Humans
  • Mutation
  • Oligopeptides / chemistry
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification


  • Actins
  • Cold Shock Proteins and Peptides
  • His-His-His-His-His-His
  • Oligopeptides
  • Recombinant Fusion Proteins
  • Histidine