Nucleic acid affinity chromatography: preparation and characterization of double-stranded RNA agarose

Protein Expr Purif. 1995 Feb;6(1):25-32. doi: 10.1006/prep.1995.1004.

Abstract

An improved method for the preparation of a double-stranded RNA-coupled resin is described. Periodate-oxidized double-stranded RNA, in the form of either purified reovirus double-stranded RNA or poly(rl). poly(rC), was covalently attached to agarose-adipic acid hydrazide. The resulting affinity resin had increased specificity for known double-stranded RNA binding proteins, as compared to a similar commercially available resin. The application of this resin in the purification and characterization of double-stranded RNA binding proteins is described.

Publication types

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

MeSH terms

  • Binding, Competitive
  • Capsid Proteins*
  • Cells, Cultured
  • Chromatography, Affinity / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Interferons / pharmacology
  • Protein Kinases / isolation & purification
  • RNA, Double-Stranded / chemistry*
  • RNA, Double-Stranded / genetics
  • RNA, Double-Stranded / metabolism
  • RNA-Binding Proteins / isolation & purification*
  • RNA-Binding Proteins / metabolism
  • Reoviridae / genetics
  • Sepharose / chemistry*
  • Viral Proteins / isolation & purification
  • Viral Proteins / metabolism

Substances

  • Capsid Proteins
  • RNA, Double-Stranded
  • RNA-Binding Proteins
  • Viral Proteins
  • sigma protein 3, Reovirus
  • Interferons
  • Sepharose
  • Protein Kinases