Selection of Highly Expressed Gene Variants in Escherichia coli Using Translationally Coupled Antibiotic Selection Markers

Research output: Chapter in Book/Report/Conference proceedingBook chapter – Annual report year: 2018Researchpeer-review

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Strategies to select highly expressed variants of a protein coding sequence are usually based on trial-and-error approaches, which are time-consuming and expensive. We address this problem using translationally coupled antibiotic resistance markers. The system requires that the target gene can be fused at the 3'-end with a translational coupling element and an antibiotic resistance gene. Highly expressed target genes can then be selected using a fast and simple whole cell survival assay in the presence of high antibiotic concentrations. Herein we show that the system can be used to select highly expressing clones from libraries sampling translation initiation sites.
Original languageEnglish
Title of host publicationSynthetic Metabolic Pathways
Volume1671
Publication date2018
Pages259-268
ISBN (Print)978-1-4939-7294-4
ISBN (Electronic)978-1-4939-7295-1
DOIs
Publication statusPublished - 2018
SeriesMethods in Molecular Biology
ISSN1064-3745
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • Antibiotic resistance, Gene expression, Library screening, Protein production optimization, Selection, Translational coupling

ID: 140018006