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

Research output: Chapter in Book/Report/Conference proceedingChapter

8 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages1-8
Number of pages8
Volume488
ISBN (Print)9781588294197
DOIs
StatePublished - 2008
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume488
ISSN (Print)10643745

Keywords

  • Affinity chromatography
  • c-src N1 exon
  • Intronic splicing silencer
  • Polypyrimidine tract binding protein
  • RNA binding proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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