Construction of an Ultra-Large Phage Display Library by Kunkel Mutagenesis and Rolling Circle Amplification.
Construction of an Ultra-Large Phage Display Library by Kunkel Mutagenesis and Rolling Circle Amplification.
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通过 Kunkel 诱变和滚环扩增构建超大型噬菌体展示库。
DOI：
10.1007/978-1-0716-3381-6_10
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Kay,BrianK
中科院分区：
文献类型：
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作者：
Huang,RenhuaR;Kierny,Michael;Volgina,Veronica;Iwashima,Makio;Miller,Christina;Kay,BrianK
An important contributor to the successful generation of recombinant affinity reagents via phage display is a large and diverse library. We describe, herein, the application of Kunkel mutagenesis and rolling circle amplification (RCA) to the construction of a 1.1 × 1011member library, with only 26 electroporations, and isolation of low- to sub-nanomolar monobodies to a number of protein targets, including human COP9 signalosome subunit 5 (COPS5), HIV-1 Rev. binding protein-like protein (HRBL), X-ray repair cross-complementing 5/6 (Ku70/80) heterodimer, the receptor-binding domain (RBD) of SARS-CoV-2, and transforming growth factor beta 1 (TGF-β1).
