Polycistronic coexpression and nondenaturing purification of histone octamers

Anal Biochem. 2012 Aug 15;427(2):190-2. doi: 10.1016/j.ab.2012.05.006. Epub 2012 May 19.

Abstract

Histone octamers are the basic building blocks of chromatin and are platforms for diverse genetic mechanisms. We report a simple method for preparing recombinant histone octamers by overexpressing all four histones from a single polycistronic vector followed by standard chromatography under native conditions. This approach reduces the time needed for the octamer preparation to a single day and should be applicable to making a variety of unmodified and modified histone octamers.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / isolation & purification
  • Bacterial Proteins / metabolism
  • Chromatin / genetics*
  • Chromatin / metabolism
  • Chromatography, Gel
  • Electrophoresis, Polyacrylamide Gel
  • Gene Expression
  • Genetic Vectors / chemistry
  • Genetic Vectors / genetics*
  • Histones / genetics*
  • Histones / isolation & purification
  • Histones / metabolism
  • Nucleosomes / genetics
  • Nucleosomes / metabolism
  • Protein Isoforms / genetics
  • Protein Isoforms / isolation & purification
  • Protein Isoforms / metabolism
  • Protein Multimerization
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • Bacterial Proteins
  • Chromatin
  • Histones
  • Nucleosomes
  • Protein Isoforms
  • Recombinant Proteins