Current strategies for mobilome research

Abstract

Mobile genetic elements (MGEs) are pivotal for bacterial evolution and adaptation, allowing shuffling of genes even between distantly related bacterial species. The study of these elements is biologically interesting as the mode of genetic propagation is kaleidoscopic and important, as MGEs are the main vehicles of the increasing bacterial antibiotic resistance that causes thousands of human deaths each year. The study of MGEs has previously focused on plasmids from individual isolates, but the revolution in sequencing technology has allowed the study of mobile genomic elements of entire communities using metagenomic approaches. The problem in using metagenomic sequencing for the study of MGEs is that plasmids and other mobile elements only comprise a small fraction of the total genetic content that are difficult to separate from chromosomal DNA based on sequence alone. The distinction between plasmid and chromosome is important as the mobility and regulation of genes largely depend on their genetic context. Several different approaches have been proposed that specifically enrich plasmid DNA from community samples. Here, we review recent approaches used to study entire plasmid pools from complex environments, and point out possible future developments for and pitfalls of these approaches. Further, we discuss the use of the PacBio long-read sequencing technology for MGE discovery.

Keywords: MGE; PacBio; horizontal gene transfer; metagenomics; mobile genetic elements; mobilome; plasmid; plasmidome.

FIGURE 1

Workflow for mobilome analysis as used in Kav et al. (2012), Li et al. (2012), Jørgensen et al. (2014), and Norman et al. (2014). Initial careful DNA extraction is followed by digestion of linear and nicked DNA with exonuclease and unspecific but selective amplification of circular DNA using multiple displacement amplification (MDA).