Spatial Mapping of Mobile Genetic Elements and their Cognate Hosts in Complex Microbiomes - Identifying the host taxon of a previously undescribed plasmid

We investigated the taxonomic association of an unknown plasmid within a plaque biofilm of a patient diagnosed with stage 3 periodontitis. We combined long- and short- read sequencing to identify a complete plasmid with minimal homology to any sequence in the RefSeq database. The plasmid carried several predicted genes for mobilization and toxin-antitoxin systems. We designed MGE-FISH probes for the plasmid and combined this MGE-FISH stain with an 18-genera HiPR-FISH panel.

Images are labeled by collection time such that the laser order for a given field of view (fov) is: 488nm Lambda, 514nm Lambda, 561nm Lambda, 633nm Airyscan, 405nm Lambda. We used Flye (https://github.com/fenderglass/Flye) to assemble the plasmid using long read Nanopore sequencing only and we used OPERA-MS (https://github.com/CSB5/OPERA-MS) to do hybrid assembly with Illumina short reads and Nanopore long reads. The assemblies are in the fasta files and the reads that map to the assemblies are in the fastq files.