20.500.12556/RUL-168324 A single vector system for tunable and homogeneous dual gene expression in Escherichia coli Expression of recombinant genes can be controlled using inducible promoters. However, the most commonly used IPTG- and arabinose-inducible promoters result in an ‘all-or-nothing’ response, leading to fully induced and uninduced bacterial subpopulations. Here, we investigate whether appropriate modifications to these promoter systems can be combined into a single vector system, enabling homogenous expression of two genes of interest that can be precisely tuned using inducer concentration. We show that modifications of positive feedback loops related to inducer uptake result in homogeneous gene expression in both the T7 lactose and pBAD arabinose systems. Furthermore, these two modified systems were combined into a single vector, pRAT-sfGFP that provides the desired tunable expression of two genes of interest. Finally, we test this single-vector system as a tool for studying two-component genetic circuits, using toxin-antitoxin modules as model systems. This novel low-copy single vector expression system opens up new possibilities for investigating the function of two-component bacterial genetic circuits. dual tunable gene expression two-component genetic circuits IPTG arabinose HigBA2 Phd/Doc toxin-antitoxin nadzorovana ekspresija dveh genov dvokomponentna genetska vezja IPTG arabinoza HigBA2 Phd/Doc toksin-antitoksin true false true Angleški jezik Slovenski jezik Neznano 2025-04-09 10:39:17 2025-04-09 10:39:18 2025-04-10 04:12:57 0000-00-00 00:00:00 2025 0 0 11 str. Vol. 15, art. 99 2 Jan. 2025 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.218 2045-2322 10.1038/s41598-024-83628-5 231910915 s41598-024-83628-5.pdf s41598-024-83628-5.pdf 1 68E630C3335516501371672B71249A2C 1ec64f339424cb4d7f390cdb0a0d24933df764f13b9bc1497edddedc33bc43fb 92f89e9e-151d-11f0-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=201588 https://www.nature.com/articles/s41598-024-83628-5 1 0c5cdf7d-151d-11f0-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=201584 Fakulteta za kemijo in kemijsko tehnologijo Biotehniška fakulteta 0 0 0