2025/08/27 by Stephanie L. Brumwell, Ariela Esmurria, Alexander Crits‐Christoph +4 · 1 voice
Biochemistry, Genetics and Molecular Biology · #Vibrio bacteria research studies #Bacterial biofilms and quorum sensing
paper · pdf · doi:10.1101/2025.08.27.672616
openalex publication_date 2025/08/27 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/15
Abstract New microbial hosts with superior phenotypes, such as fast growth, are attractive for research and biotechnology, but often lack systematic evaluation of functional genetic parts. A reference set of working plasmids would increase reproducibility and encourage use of these hosts. Here, we use the POSSUM toolkit, a collection of 23 origins-of-replication and 6 antibiotic markers, to identify functional genetic parts for strains of Vibrio natriegens . We applied this to the wild-type strain ATCC 14048 and an engineered variant NBx CyClone™, evaluating 414 combinations of origins of replication and antibiotic selection conditions. We show that both strains support five replicons (pNG2, pSa, pSC101ts, p15A, and RSF1010) with NBx CyClone™ supporting an extra replicon RK2. The assay can be performed in under a week and is compatible with multiple DNA delivery methods. This work demonstrates the feasibility of rapidly establishing reference information to accelerate the adoption of new microbial hosts.