Method of mRNA Display for Selecting Peptides That Bind with Target Enzyme RNA

In vitro display techniques are useful tools for selecting novel ligands that bind to target biomolecules. In this paper, mRNA display method was used to select peptides that bind thymidylate synthase RNA with high affinity from a peptide library. We provide a detailed analysis of the critical steps of an mRNA-display-based selection, including the generation of an mRNA-displayed library, formation of RNA-peptide fusions, and selection of RNA affinity peptides. After optimization of the experimental steps, the efficiency of RNA-linker purification and the reverse transcription was Improved, and the unspecified binding peptides were removed. Our study could facilitate the use of mRNA display in the field of biotechnology, medicine and proteomics.