Isolation of a sequence-specific RNA binding protein, polypyrimidine tract binding protein, using RNA affinity chromatography

Shalini Sharma

doi:10.1007/978-1-60327-475-3_1

Isolation of a sequence-specific RNA binding protein, polypyrimidine tract binding protein, using RNA affinity chromatography

Methods Mol Biol. 2008:488:1-8. doi: 10.1007/978-1-60327-475-3_1.

Author

Shalini Sharma¹

Affiliation

¹ Howard Hughes Medical Institute, University of California at Los Angeles, Los Angeles, California, USA.

PMID: 18982280
DOI: 10.1007/978-1-60327-475-3_1

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromatography, Affinity / methods*
Polypyrimidine Tract-Binding Protein / chemistry
Polypyrimidine Tract-Binding Protein / isolation & purification*
RNA / chemistry*
RNA-Binding Proteins / chemistry
RNA-Binding Proteins / isolation & purification

Substances

RNA-Binding Proteins
Polypyrimidine Tract-Binding Protein
RNA

Grants and funding

Howard Hughes Medical Institute/United States