Isolation of a sequence-specific RNA binding protein, polypyrimidine tract binding protein, using RNA affinity chromatography.
Many important cellular processes are mediated by sequence-specific RNA binding proteins, and it is often necessary to purify these proteins. When the RNA binding site is known, it is convenient to use this RNA as a matrix for affinity purification. The intronic splicing silencer (ISS) element present upstream of the N1 exon of the c-src pre-mRNA is a high-affinity binding site for the polypyrimidine tract binding protein (PTB). Using a 5'-biotinylated ISS RNA and PTB as an example, I describe a one-step method for affinity chromatography of RNA binding proteins from nuclear extracts.
Sharma
Shalini
S
Howard Hughes Medical Institute, University of California at Los Angeles, Los Angeles, California, USA.
D002846:Chromatography, Affinity / Q000379:methods*; D038941:Polypyrimidine Tract-Binding Protein / Q000737:chemistry / Q000302:isolation & purification*; D012313:RNA / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification
D016601:RNA-Binding Proteins; D038941:Polypyrimidine Tract-Binding Protein; D012313:RNA