Wang Rong, Cotten Steve W, Liu Rihe
Eshelman School of Pharmacy and Carolina Center for Genome Sciences, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Methods Mol Biol. 2012;805:87-100. doi: 10.1007/978-1-61779-379-0_6.
mRNA display is a powerful technique that allows for covalent coupling of a translated protein with its coding mRNA. The resulting conjugation between genotype and phenotype can be used for the efficient selection and identification of peptides or proteins with desired properties from an mRNA-displayed peptide or protein library with high diversity. This protocol outlines the principle of mRNA display and the detailed procedures for the synthesis of mRNA-protein fusions. Some special considerations for library construction, generation, and purification are discussed.
信使核糖核酸展示是一种强大的技术,它能够使翻译后的蛋白质与其编码的信使核糖核酸进行共价偶联。基因型与表型之间产生的这种共轭作用可用于从具有高度多样性的信使核糖核酸展示肽或蛋白质文库中高效筛选和鉴定具有所需特性的肽或蛋白质。本方案概述了信使核糖核酸展示的原理以及信使核糖核酸-蛋白质融合体合成的详细步骤。还讨论了文库构建、生成和纯化的一些特殊注意事项。