He Mingyue, Edwards Bryan M, Kastelic Damjana, Taussig Michael J
The Inositide Laboratory, The Babraham Institute, Cambridge, UK.
Methods Mol Biol. 2012;805:75-85. doi: 10.1007/978-1-61779-379-0_5.
Ribosome display is a cell-free display technology for in vitro selection and optimisation of proteins from large diversified libraries. It operates through the formation of stable protein-ribosome-mRNA (PRM) complexes and selection of ligand-binding proteins, followed by DNA recovery from the selected genetic information. Both prokaryotic and eukaryotic ribosome display systems have been developed. In this chapter, we describe the eukaryotic rabbit reticulocyte method in which a distinct in situ single-primer RT-PCR procedure is used to recover DNA from the selected PRM complexes without the need for prior disruption of the ribosome.
核糖体展示是一种无细胞展示技术，用于从大型多样化文库中体外筛选和优化蛋白质。它通过形成稳定的蛋白质-核糖体-信使核糖核酸（PRM）复合物并选择配体结合蛋白来运作，随后从所选遗传信息中回收DNA。原核和真核核糖体展示系统均已得到开发。在本章中，我们描述了真核兔网织红细胞方法，其中使用独特的原位单引物逆转录-聚合酶链反应（RT-PCR）程序从所选的PRM复合物中回收DNA，而无需事先破坏核糖体。
