用户自定义单锅诱变使用未扩增寡核苷酸池。

IF 2.6 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Protein Engineering Design & Selection Pub Date : 2019-09-10 DOI:10.1093/protein/gzz013

Angélica V Medina-Cucurella, Paul J Steiner, Matthew S Faber, Jesús Beltrán, Alexandra N Borelli, Monica B Kirby, Sean R Cutler, Timothy A Whitehead

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引用次数: 17

摘要

用户定义的诱变文库是应用蛋白质工程工作流程的基础。本研究表明，未扩增的寡核苷酸库可用于从质粒DNA制备位点饱和诱变文库，几乎完全覆盖所需突变和少数脱靶突变。我们发现，与单独合成的退化寡聚相比，寡聚池产生更高质量的库。我们还表明，多个库可以多路复用到一个oligo池中，从而使多个库的准备成本更低，更方便。我们为自动寡核苷酸池设计提供软件，可以为饱和或集中的库生成诱变寡核苷酸。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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User-defined single pot mutagenesis using unamplified oligo pools.

User-defined mutagenic libraries are fundamental for applied protein engineering workflows. Here we show that unamplified oligo pools can be used to prepare site saturation mutagenesis libraries from plasmid DNA with near-complete coverage of desired mutations and few off-target mutations. We find that oligo pools yield higher quality libraries when compared to individually synthesized degenerate oligos. We also show that multiple libraries can be multiplexed into a single oligo pool, making preparation of multiple libraries less expensive and more convenient. We provide software for automatic oligo pool design that can generate mutagenic oligos for saturating or focused libraries.

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来源期刊

Protein Engineering Design & Selection 生物-生化与分子生物学

CiteScore

3.30

自引率

4.20%

发文量

审稿时长

6-12 weeks

期刊介绍： Protein Engineering, Design and Selection (PEDS) publishes high-quality research papers and review articles relevant to the engineering, design and selection of proteins for use in biotechnology and therapy, and for understanding the fundamental link between protein sequence, structure, dynamics, function, and evolution.