Selection of RNA-binding Peptides Using mRNA-peptide Fusions

Methods. 2001 Mar;23(3):287-93. doi: 10.1006/meth.2000.1139.

Abstract

We have been working to apply in vitro selection to isolate novel RNA-binding peptides. To do this, we use mRNA-protein fusions, peptides covalently attached to their own mRNA. Here, we report selection protocols developed using the arginine-rich domain of bacteriophage lambda-N protein and its binding target, the boxB RNA. Systematic investigation of possible paths for a selection round has allowed us to design a reliable and efficient protocol to enrich RNA-binding peptides from nonfunctional members of a complex mixture. The protocols we have developed should greatly facilitate the isolation of new molecules using the fusion system.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA, Complementary / metabolism
  • Genetic Techniques*
  • Peptides*
  • Protein Biosynthesis
  • Protein Structure, Tertiary
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / metabolism*
  • Recombinant Fusion Proteins / metabolism*

Substances

  • DNA, Complementary
  • Peptides
  • RNA, Messenger
  • RNA-Binding Proteins
  • Recombinant Fusion Proteins