Translating ribosome affinity purification (TRAP) for cell-specific translation profiling in developing flowers

Methods Mol Biol. 2014:1110:323-8. doi: 10.1007/978-1-4614-9408-9_18.

Abstract

The development of a multicellular organism is accompanied by cell differentiation. In fact, many biological processes have cell specificity, such that distinct cell types respond differently to endogenous or environmental cues. To obtain cell-specific gene expression profiles, translating ribosome affinity purification (TRAP) has been developed to label polysomes containing translating mRNAs in genetically defined cell types. Here, we describe the immunopurification of epitope-labeled polysomes and associated RNAs from target cell types. TRAP has the additional advantage of obtaining only translating mRNAs, which are a better proxy to the proteome than a standard mRNA preparation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / growth & development
  • Flowers / genetics*
  • Flowers / growth & development*
  • Immunoprecipitation / methods*
  • Microspheres
  • Polyribosomes / genetics
  • Protein Biosynthesis*
  • RNA, Plant / genetics
  • RNA, Plant / isolation & purification
  • Ribosomes / genetics*
  • Sepharose / chemistry

Substances

  • RNA, Plant
  • Sepharose