title: In vitro selection of proteins with desired characteristics using mRNA-display.
abstract: mRNA-display is an amplification-based, iterative rounds of in vitro protein selection technique that circumvents a number of difficulties associated with yeast two-hybrid and phage display. Because of the covalent linkage between the genotype and the phenotype, mRNA-display provides a powerful means for reading and amplifying a peptide or protein sequence after it has been selected from a library with very high diversity. The purpose of this article is to provide a summary of the field and practical framework of mRNA-display-based selections. We summarize the advantages and limitations of selections using mRNA-display as well as the recent applications, namely, the identification of novel affinity reagents, target-binding partners, and enzyme substrates from synthetic peptide or natural proteome libraries. Practically, we provide a detailed procedure for performing mRNA-display-based selections with the aim of identifying protease substrates and binding partners of a target protein. Furthermore, we describe how to confirm the function of the selected protein sequences by biochemical assays and bioinformatic tools.
authors.affiliation: Division of Human Genetics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA. alexander.valencia@cchmc.org
authors.forename: C Alexander
authors.initials: CA
authors.lastname: Valencia
authors.forename: Jianwei
authors.initials: J
authors.lastname: Zou
authors.forename: Rihe
authors.initials: R
authors.lastname: Liu
mesh_terms: D006863:Hydrogen-Ion Concentration; D019151:Peptide Library; D011485:Protein Binding; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*
chemical_list: D019151:Peptide Library; D011506:Proteins; D012333:RNA, Messenger