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mRNA‐Display‐Based Selections for Proteins with Desired Functions: A Protease‐Substrate Case Study
Author(s) -
Valencia C. Alexander,
Cotten Steven W.,
Dong Biao,
Liu Rihe
Publication year - 2008
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp070473a
Subject(s) - phage display , proteome , computational biology , messenger rna , peptide library , biology , translation (biology) , protease , selection (genetic algorithm) , peptide , bioinformatics , genetics , peptide sequence , biochemistry , enzyme , computer science , gene , artificial intelligence
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 a diversity in the range of 10 12 –10 13 . In this paper, we briefly review the recent progress in using mRNA‐display to identify affinity reagents, binding partners, and enzyme substrates from synthetic peptide or natural proteome libraries. To facilitate the use of mRNA‐display in research laboratories without previous experience, we provide a detailed analysis of the critical steps of an mRNA‐display‐based selection in a case study for the identification of the natural substrates of caspases, including the generation of an mRNA‐displayed proteome library, removal of abundant sequences, and selection of proteins with desired functions. The advantages and technical limitations of mRNA‐display as a general peptide or protein selection tool are also addressed.