Random transposon mutagenesis of Verrucomicrobium spinosum DSM 4136(T).

The Verrucomicrobia are a bacterial group of growing interest due to their environmental ubiquity as free-living and host-associated microbes. They also exhibit an unusual compartmentalized cell plan, shared with members of neighboring phyla that include the Planctomycete bacteria. However, Verrucomicrobia are currently difficult to study, due to a lack of available genetic tools that would permit robust testing of hypotheses formulated from ecological and genomic data. To our knowledge, there are no published studies describing the transformation of exogenous DNA into any members of the Verrucomicrobia (or the neighboring phylum containing Planctomycetes). Here, we present a procedure for the transformation of DNA into Verrucomicrobium spinosum DSM 4136(T) via electroporation and the first description of a random transposon mutant library in this organism. We anticipate that this approach could be applied successfully to other Verrucomicrobia, providing opportunities to test the role of predicted gene function in ecological interactions and identify genes associated with the distinctive Planctomycete-Verrucomicrobial cell plan.