Expression and Purification of RNA-Protein Complexes in Escherichia coli

Methods Mol Biol. 2015:1316:25-31. doi: 10.1007/978-1-4939-2730-2_3.

Abstract

For structural, biochemical or pharmacological studies, it is required to have pure RNA in large quantities. We previously devised a generic approach that allows efficient in vivo expression of recombinant RNA in Escherichia coli. We have extended the "tRNA scaffold" method to RNA/protein co-expression in order to express and purify RNA by affinity in native condition. As a proof-of-concept, we present the expression and the purification of the AtRNA-mala in complex with the MS2 coat protein.

MeSH terms

  • Escherichia coli / genetics*
  • Escherichia coli / metabolism*
  • Gene Expression*
  • Nucleic Acid Conformation
  • Protein Binding
  • Proteins / genetics*
  • Proteins / isolation & purification*
  • Proteins / metabolism
  • RNA / chemistry
  • RNA / genetics*
  • RNA / isolation & purification*
  • RNA / metabolism
  • RNA, Transfer / chemistry
  • RNA, Transfer / genetics
  • RNA, Transfer / isolation & purification
  • RNA, Transfer / metabolism

Substances

  • Proteins
  • RNA
  • RNA, Transfer