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Plasmids are a highly effective means with which genetic traits that influence human health, such as virulence and antibiotic resistance, are disseminated through bacterial populations. The IncX-family is a hitherto sparsely populated group of plasmids that are able to thrive within  Enterobacteriaceae . In this study, a replicon-centric screening method was used to locate strains from wastewater sludge containing plasmids belonging to the IncX-family. A transposon aided plasmid capture method was then employed to transport IncX-plasmids from their original hosts (and co-hosted plasmids) into a laboratory strain ( Escherichia coli  Genehogs®) for further study. The nucleotide sequences of the three newly isolated IncX-plasmids (pLN126_33, pMO17_54, pMO440_54) and the hitherto un-sequenced type-plasmid R485 revealed a remarkable occurrence of whole or partial gene cassettes that promote biofilm-formation in  Klebsiella pneumonia  or  E. coli , in all four instances. Two of the plasmids (R485 and pLN126_33) were shown to directly induce biofilm formation in a crystal violet retention assay in  E. coli . Sequence comparison revealed that all plasmid-borne forms of the type 3 fimbriae encoding gene cassette  mrkABCDF  were variations of a composite transposon Tn 6011  first described in the  E. coli  IncX plasmid pOLA52. In conclusion, IncX-plasmids isolated from  Enterobacteriaceae  over almost 40 years and on three different continents have all been shown to carry a type 3 fimbriae gene cassette  mrkABCDF  stemming from pathogenic  K. pneumoniae . Apart from contributing general knowledge about IncX-plasmids, this study also suggests an apparent ubiquity of a mobile form of an important virulence factor and is an illuminating example of the recruitment, evolution and dissemination of genetic traits through plasmid-mediated horizontal gene transfer.

Sequencing of IncX-Plasmids Suggests Ubiquity of Mobile Forms of a Biofilm-Promoting Gene Cassette Recruited from Klebsiella pneumoniae