用于肽和蛋白质体外筛选的RNA-肽融合体
RNA-peptide fusions for the in vitro selection of peptides and proteins.
作者信息
Roberts R W, Szostak J W
机构信息
Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114, USA.
出版信息
Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12297-302. doi: 10.1073/pnas.94.23.12297.
Abstract
Covalent fusions between an mRNA and the peptide or protein that it encodes can be generated by in vitro translation of synthetic mRNAs that carry puromycin, a peptidyl acceptor antibiotic, at their 3' end. The stable linkage between the informational (nucleic acid) and functional (peptide) domains of the resulting joint molecules allows a specific mRNA to be enriched from a complex mixture of mRNAs based on the properties of its encoded peptide. Fusions between a synthetic mRNA and its encoded myc epitope peptide have been enriched from a pool of random sequence mRNA-peptide fusions by immunoprecipitation. Covalent RNA-peptide fusions should provide an additional route to the in vitro selection and directed evolution of proteins.
摘要
信使核糖核酸（mRNA）与其编码的肽或蛋白质之间的共价融合，可以通过体外翻译3'端携带嘌呤霉素（一种肽基受体抗生素）的合成mRNA来实现。所得连接分子的信息（核酸）和功能（肽）结构域之间的稳定连接，使得特定的mRNA能够基于其编码肽的特性，从复杂的mRNA混合物中富集出来。通过免疫沉淀，已从随机序列mRNA-肽融合文库中富集了合成mRNA与其编码的Myc表位肽之间的融合物。共价RNA-肽融合应为蛋白质的体外筛选和定向进化提供一条额外途径。
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