title: Terminator Operon Reporter: combining a transcription termination switch with reporter technology for improved gene synthesis and synthetic biology applications.
abstract: Synthetic biology is characterized by the development of novel and powerful DNA fabrication methods and by the application of engineering principles to biology. The current study describes Terminator Operon Reporter (TOR), a new gene assembly technology based on the conditional activation of a reporter gene in response to sequence errors occurring at the assembly stage of the synthetic element. These errors are monitored by a transcription terminator that is placed between the synthetic gene and reporter gene. Switching of this terminator between active and inactive states dictates the transcription status of the downstream reporter gene to provide a rapid and facile readout of the accuracy of synthetic assembly. Designed specifically and uniquely for the synthesis of protein coding genes in bacteria, TOR allows the rapid and cost-effective fabrication of synthetic constructs by employing oligonucleotides at the most basic purification level (desalted) and without the need for costly and time-consuming post-synthesis correction methods. Thus, TOR streamlines gene assembly approaches, which are central to the future development of synthetic biology.
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Zampini
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Luis A J
authors.initials: LA
authors.lastname: Mur
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Pauline
authors.initials: P
authors.lastname: Rees Stevens
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Justin A
authors.initials: JA
authors.lastname: Pachebat
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: C James
authors.initials: CJ
authors.lastname: Newbold
authors.affiliation: Faculty of Life Sciences, University of Manchester, Manchester M13 9PL, UK.
authors.forename: Finbarr
authors.initials: F
authors.lastname: Hayes
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Alison
authors.initials: A
authors.lastname: Kingston-Smith
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D009876:Operon; D058615:Synthetic Biology; D061806:Transcription Termination, Genetic