Directed evolution of transcriptional switches using dual-selector systems

Yuki Kimura, Daisuke Umeno*

*この研究の対応する著者

研究成果: Chapter

抄録

Genetic switches provide core components of gene expression induction systems, genetic circuits, and metabolite sensors, including those for high-throughput screening and selection of enzyme functions. However, it is rare that natural transcription factors meet the required specifications for each application. Fortunately, the directed evolution of transcription switches is a straightforward process, given that the two states of genetic switches (on and off) are both selectable, using a wide range of positive and negative selection tools developed in the field of molecular genetics. On/off-state selections based on bactericidal mechanism allow greatly accelerate the entire process. The key to success is finding the selection conditions that minimize false-positive and false-negative clones as with any directed evolution experiment. We introduce a reliable and automatable directed evolution platform to rapidly evolve genetic switches with desired specifications in this chapter. Highlighting the importance and ease of screening for selection conditions, we demonstrate how selection conditions influence the resultant population of the selected pools.

本文言語English
ホスト出版物のタイトルEnzyme Engineering and Evolution
ホスト出版物のサブタイトルSpecific Enzyme Applications
編集者Dan S. Tawfik
出版社Academic Press Inc.
ページ191-207
ページ数17
ISBN(印刷版)9780128244319
DOI
出版ステータスPublished - 2020
外部発表はい

出版物シリーズ

名前Methods in Enzymology
644
ISSN(印刷版)0076-6879
ISSN(電子版)1557-7988

ASJC Scopus subject areas

  • 生化学
  • 分子生物学

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