ArticleNucleic acids research2016
A part toolbox to tune genetic expression in Bacillus subtilis.
Article in Nucleic acids research, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 112 papers.
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Who cites it
112 citing papers in PubMed.
- Identification of integration site library for the development of plasmid-free microbial cell factory inSynthetic and systems biotechnology · 2027Article
- An inducer-independent, single-plasmid CRISPR-Cas9 system for genome editing in Bacillus species.Applied microbiology and biotechnology · 2026Article
- RATEX: A Scalable RNA-Based Platform for Logical and Multi-Layered Cellular Programming.Angewandte Chemie (International ed. in English) · 2026Article
- A multi-tetracycline responsive induction system for gene expression inApplied and environmental microbiology · 2026Article
- Rhodo-Box: A Synthetic Biology Toolbox to Facilitate Metabolic Engineering ofACS synthetic biology · 2026Article
- Origin of replication discovery for environmentally isolatediScience · 2026Article
- Synthetic Biology of Plants and Microbes for Agriculture, Environment, and Future Applications.Chemical reviews · 2026Review
- Deep Generative Model-Driven Design of Microbial Synthetic Promoters.Journal of microbiology and biotechnology · 2025Review
- A versatile plug and play bacterial chassis based on a fast-growing Mycoplasma species.Nucleic acids research · 2025Article
- Construction of aACS omega · 2025Article
- A Multi-Tetracycline Responsive Induction System for Gene Expression inbioRxiv : the preprint server for biology · 2025Article
- Engineering Bacillus subtilis for high-value bioproduction: recent advances and applications.Microbial cell factories · 2025Review
- Real-time nanoscale investigation of spore coat assembly in Bacillus subtilis.Communications biology · 2025Article
- Expanding genetic engineering capabilities in Vibrio natriegens with the Vnat Collection.Nucleic acids research · 2025Article
- The mutational landscape of Bacillus subtilis conditional hypermutators shows how proofreading skews DNA polymerase error rates.Nucleic acids research · 2025Article
- Optogenetic Control of B. subtilis Gene Expression Using the CcaSR System.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Exploring the Promoter Generation and Prediction ofInternational journal of molecular sciences · 2024Article
- A novel interpretable deep learning-based computational framework designed synthetic enhancers with broad cross-species activity.Nucleic acids research · 2024Article
- A promoter-RBS library for fine-tuning gene expression inApplied and environmental microbiology · 2024Article
- A role for the S4-domain containing protein YlmH in ribosome-associated quality control in Bacillus subtilis.Nucleic acids research · 2024Article
52 more citing papers are in PubMed but not listed here.
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Libraries of well-characterised components regulating gene expression levels are essential to many synthetic biology applications. While widely available for the Gram-negative model bacterium Escherichia coli, such libraries are lacking for the Gram-positive model Bacillus subtilis, a key organism for basic research and biotechnological applications. Here, we engineered a genetic toolbox comprising libraries of promoters, Ribosome Binding Sites (RBS), and protein degradation tags to precisely tune gene expression in B. subtilis We first designed a modular Expression Operating Unit (EOU) facilitating parts assembly and modifications and providing a standard genetic context for gene circuits implementation. We then selected native, constitutive promoters of B. subtilis and efficient RBS sequences from which we engineered three promoters and three RBS sequence libraries exhibiting ∼14 000-fold dynamic range in gene expression levels. We also designed a collection of SsrA proteolysis tags of variable strength. Finally, by using fluorescence fluctuation methods coupled with two-photon microscopy, we quantified the absolute concentration of GFP in a subset of strains from the library. Our complete promoters and RBS sequences library comprising over 135 constructs enables tuning of GFP concentration over five orders of magnitude, from 0.05 to 700 μM. This toolbox of regulatory components will support many research and engineering applications in B. subtilis.
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