{"id":"doi:10.1016/j.ymben.2016.09.008","kind":"article","title":"Development of a Bacillus subtilis cell-free transcription-translation system for prototyping regulatory elements","authors":["Richard J. R. Kelwick","Alexander J. Webb","James T. MacDonald","Paul S. Freemont"],"author_count":4,"year":2016,"published":"2016-10-06","venue":"Metabolic Engineering","cited_by":135,"doi":"10.1016/j.ymben.2016.09.008","arxiv_id":null,"nct_id":null,"source_url":"https://doi.org/10.1016/j.ymben.2016.09.008","archive_url":"https://space-kitty.com/papers/doi:10.1016/j.ymben.2016.09.008","updated":null,"pmid":"27697563","pmcid":null,"openalex_id":"W2528808089","pdf_url":null,"abstract":"Cell-free transcription-translation systems were originally applied towards in vitro protein production. More recently, synthetic biology is enabling these systems to be used within a systematic design context for prototyping DNA regulatory elements, genetic logic circuits and biosynthetic pathways . The Gram-positive soil bacterium , Bacillus subtilis , is an established model organism of industrial importance. To this end, we developed several B. subtilis -based cell-free systems. Our improved B. subtilis WB800N-based system was capable of producing 0.8 µM GFP, which gave a ~72x fold-improvement when compared with a B. subtilis 168 cell-free system. Our improved system was applied towards the prototyping of a B. subtilis promoter library in which we engineered several promoters, derived from the wild-type P grac ( σ A) promoter, that display a range of comparable in vitro and in vivo transcriptional activities. Additionally, we demonstrate the cell-free characterisation of an inducible expression system , and the activity of a model enzyme - renilla luciferase .","abstract_license":"CC-BY (publisher license, via OpenAlex)","license":"cc-by","is_open_access":true,"retracted":false,"peer_reviewed":null,"categories":["Microbial Metabolic Engineering and Bioproduction"],"details":null,"provenance":"openalex","attribution":"Abstract of \"Development of a Bacillus subtilis cell-free transcription-translation system for prototyping regulatory elements\" by Richard J. R. Kelwick, Alexander J. Webb, James T. MacDonald et al., https://doi.org/10.1016/j.ymben.2016.09.008, used under its Creative Commons Attribution license (CC-BY); markup removed, otherwise unmodified.","topics":[{"slug":"synthetic-biology","name":"Synthetic Biology & Genome Engineering","domain_slug":"biotech"}],"cited_in_dossiers":[]}