Generating a Collection of Single-Gene Deletions in the eps Operon in Bacillus subtilis Strain NCIB 3610
In the environment, bacteria mainly exist in multicellular, surface-adhered communities called biofilms. One key feature of the biofilms is a self-produced extracellular matrix that embeds cells within a structured community and contributes to their adhesion, organization, and retention. The matrix of most characterized biofilms contains three primary components: polysaccharides, protein, and…
In Bacillus subtilis strain NCIB 3610, a collection of marker-less, in-frame deletions targeting each gene in the epsA-O operon was created. The epsA-O operon encodes proteins that coordinate the assembly, export, and polymerization of exopolysaccharide (EPS) oligosaccharide subunits. The three primary components of most characterized biofilms are polysaccharides, protein, and extracellular DNA (eDNA). Upon biofilm induction, B. subtilis produces and secretes a polysaccharide known as exopolysaccharide (EPS).
The aim of this study was to investigate the contribution of each eps gene to EPS production and biofilm-associated phenotypes. To achieve this, the Cre system was used to generate a collection of single-gene deletions in B. subtilis strain NCIB 3610. Biofilm phenotypes of the resulting mutants were then assessed to determine the importance of each gene in biofilm development.
The epsA-O single-gene deletion collection developed in this study serves to systematically assess the role of each gene in EPS production and biofilm-associated phenotypes. This collection, combined with existing knowledge of EPS structure, will help clarify the roles of individual genes in EPS biosynthesis. Moreover, this approach can be easily adapted for systematic analyses of other large, complex operons in diverse bacterial species.
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