A nucleoside kinase as a dual selector for genetic switches and circuits

Yohei Tashiro, Hiroki Fukutomi, Kei Terakubo, Kyoichi Saito, Daisuke Umeno

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

The development of genetic switches and their integrated forms (genetic circuits) with desired specifications/functions is key for success in synthetic biology. Due to the difficulty in rational design, genetic switches and circuits with desirable specifications are mostly obtained by directed evolution. Based on a virus-derived nucleotide kinase as a single-gene dual selector, we constructed a robust, efficient and stringent selection system for genetic switches. This method exhibited unprecedented enrichment efficacy (>30000-fold) of functional switches from non-functional ones in a single selection cycle. In addition, negative (OFF) selection was exceptionally stringent, allowing the rapid and efficient selection of non-leaky from leaky circuits.

Original languageEnglish
Pages (from-to)e12
JournalNucleic acids research
Volume39
Issue number3
DOIs
Publication statusPublished - 2011 Feb
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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