Dual genetic selection of synthetic riboswitches in Escherichia coli

Yoko Nomura, Yohei Yokobayashi

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

This chapter describes a genetic selection strategy to engineer synthetic riboswitches that can chemically regulate gene expression in Escherichia coli. Riboswitch libraries are constructed by randomizing the nucleotides that potentially comprise an expression platform and fused to the hybrid selection/screening marker tetA-gfpuv. Iterative ON and OFF selections are performed under appropriate conditions that favor the survival or the growth of the cells harboring the desired riboswitches. After the selection, rapid screening of individual riboswitch clones is performed by measuring GFPuv fluorescence without subcloning. This optimized dual genetic selection strategy can be used to rapidly develop synthetic riboswitches without detailed computational design or structural knowledge.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages131-140
Number of pages10
Volume1111
ISBN (Print)9781627037549
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1111
ISSN (Print)10643745

Fingerprint

Riboswitch
Genetic Selection
Escherichia coli
Nucleotides
Clone Cells
Fluorescence
Gene Expression
Growth

Keywords

  • Aptamer
  • Gene regulation
  • Riboswitch
  • RNA engineering
  • Translational regulation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Nomura, Y., & Yokobayashi, Y. (2014). Dual genetic selection of synthetic riboswitches in Escherichia coli. In Methods in Molecular Biology (Vol. 1111, pp. 131-140). (Methods in Molecular Biology; Vol. 1111). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-755-6-9

Dual genetic selection of synthetic riboswitches in Escherichia coli. / Nomura, Yoko; Yokobayashi, Yohei.

Methods in Molecular Biology. Vol. 1111 Humana Press Inc., 2014. p. 131-140 (Methods in Molecular Biology; Vol. 1111).

Research output: Chapter in Book/Report/Conference proceedingChapter

Nomura, Y & Yokobayashi, Y 2014, Dual genetic selection of synthetic riboswitches in Escherichia coli. in Methods in Molecular Biology. vol. 1111, Methods in Molecular Biology, vol. 1111, Humana Press Inc., pp. 131-140. https://doi.org/10.1007/978-1-62703-755-6-9
Nomura Y, Yokobayashi Y. Dual genetic selection of synthetic riboswitches in Escherichia coli. In Methods in Molecular Biology. Vol. 1111. Humana Press Inc. 2014. p. 131-140. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-755-6-9
Nomura, Yoko ; Yokobayashi, Yohei. / Dual genetic selection of synthetic riboswitches in Escherichia coli. Methods in Molecular Biology. Vol. 1111 Humana Press Inc., 2014. pp. 131-140 (Methods in Molecular Biology).
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