mRNA-display-based selections for proteins with desired functions: a protease-substrate case study.
mRNA-display-based selections for proteins with desired functions: a protease-substrate case study.
复制标题
基于 mRNA 展示的具有所需功能的蛋白质的选择：蛋白酶底物案例研究。
DOI：
10.1021/bp070473a
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发表时间：
2008
影响因子：
2.9
通讯作者：
Liu,Rihe
中科院分区：
文献类型：
--
作者：
Valencia,CAlexander;Cotten,StevenW;Dong,Biao;Liu,Rihe
mRNA‐display is an amplification‐based, iterative rounds ofin vitroprotein 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 1012–1013. 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.
