Designing efficient genetic code expansion in Bacillus subtilis to gain biological insights
Year: 2021
Type: article
Abstract: Bacillus subtilis is a model gram-positive bacterium, commonly used to explore questions across bacterial cell biology and for industrial uses. To enable greater understanding and control of proteins in B. subtilis, here we report broad and efficient genetic code expansion in B. subtilis by incorporating 20 distinct non-standard amino acids within proteins using 3 different families of genetic code expansion systems and two choices of codons. We use these systems to achieve click-labelling, photo-crosslinking, and translational titration. These tools allow us to demonstrate differences between E. coli and B. subtilis stop codon suppression, validate a predicted protein-protein binding interface, and begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo. We expect that the establishment of this simple and easily accessible chemical biology system in B. subtilis will help uncover an abundance of biological insights and aid genetic code expansion in other organisms.
Source: Nature Communications
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $5,560
Awards: G1739570012, G1962169235, G222768804, G226688381, MICROBIAL ECOLOGY, PROTEOGENOMICS AND COMPUTATIONAL OPTIMA, MICROBIAL ECOLOGY, PROTEOGENO-MICS & COMPUTATIONAL OPTIMA, G5520316217, Collaborative Research: Enabling control of Bacillus subtilis growth using non-standard amino acids, G6322695936, Membrane-associated DNA transport machines in Bacillus subtilis, G8474615091, Graduate Research Fellowship Program (GRFP), G9000091595, G9000102033, G9000176420
OpenAlex ID: W3200853817