Nucleic acid affinity chromatography: preparation and characterization of double-stranded RNA agarose.
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.
Langland
J O
JO
Department of Microbiology, Arizona State University, Tempe 85287-2701, USA.
Pettiford
S M
SM
Jacobs
B L
BL
D001667:Binding, Competitive; D036022:Capsid Proteins; D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D007372:Interferons / Q000494:pharmacology; D011494:Protein Kinases / Q000302:isolation & purification; D012330:RNA, Double-Stranded / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D012087:Reoviridae / Q000235:genetics; D012685:Sepharose / Q000737:chemistry*; D014764:Viral Proteins / Q000302:isolation & purification / Q000378:metabolism
D036022:Capsid Proteins; D012330:RNA, Double-Stranded; D016601:RNA-Binding Proteins; D014764:Viral Proteins; C053179:sigma protein 3, Reovirus; D007372:Interferons; D012685:Sepharose; D011494:Protein Kinases