Abstract
The rapid prototyping and optimization of plasmid-based recombinant gene expression is one of the key steps in the development of bioengineered bacterial systems. Often, multiple genes or gene modules need to be coexpressed, and for this purpose compatible, inducible plasmid systems have been developed. However, inducible expression systems are not favored in industrial processes, due to their prohibitive cost, and consequently the conversion to constitutive expression systems is often desired. Here we present a set of constitutive-expression plasmids for this purpose, which were benchmarked using fluorescent reporter genes. To further facilitate the conversion between inducible and constitutive expression systems, we developed SelProm, a design tool that serves as a parts repository of plasmid expression strength and predicts portability rules between constitutive and inducible plasmids through model comparison and machine learning. The SelProm tool is freely available at http://selprom.synbiochem.co.uk .
Highlights
The rapid prototyping and optimization of plasmid-based recombinant gene expression is one of the key steps in the development of bioengineered bacterial systems
A large set of compatible, well-characterized inducible expression plasmids was created by Lee and co-workers,[5] which allows the informed coexpression of multiple genes/pathways with a range of inducer compounds, providing a valuable and enabling resource for the synthetic biology community
Despite there being a large number of characterized constitutive promoters, there is no database which holds their relative strengths in the context of a plasmid or that allows the direct comparison of inducible and constitutive expression systems
Summary
Letter inducible plasmids, were benchmarked and the resulting data were used to build a dynamic database, SelProm, which stores the performance (relative strength), dynamic response and stability of gene expression, along with standardized data sheets for each plasmid. The collated experimental data were used to train statistical learning-based algorithms to predict relative promoter strength for untested plasmids/ conditions in both BglBrick plasmid collections. The SelProm design tool integrates functionality to allow the comparison of inducible promoter expression levels to those of constitutive promoters across a range of conditions (strain, media, plasmid). This tool will enhance the portability between prototyped inducible plasmids and constitutive expression plasmids, addressing some of the most common challenges in bacterial strain engineering
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