A nucleoside kinase as a dual selector for genetic switches and circuits.
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 (>30,000-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.
Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Chiba University, 1-33 Yayoi-Cyo, Inage-ku, Chiba 263-8522, Japan.
Yohei
Y
Tashiro
Hiroki
H
Fukutomi
Kei
K
Terakubo
Kyoichi
K
Saito
Daisuke
D
Umeno
D053263:Gene Regulatory Networks; D005818:Genetic Engineering / Q000379:methods*; D008957:Models, Genetic; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000378:metabolism*
D013937:Thymidine Kinase