Metagenomic approaches are being use to investigate many diverse ecosystems and reveal many novel functions therein. However these methods can... » More

Metagenomic approaches are being use to investigate many diverse ecosystems and reveal many novel functions therein. However these methods can frequently pass over the DNA which we find in the mobile metagenome or mobilome. Much of this DNA is not easily cloned into fosmids or BACs for functional analysis and DNA based metagenomics projects do not always provide sufficient information to determine from where the sequence arose, genome or mobilome. Couple these limitations to the fact that the current approaches for isolating mobile genetic elements (MGEs), especially plasmids, are either biased to the organisms which are able to grow on laboratory media (endogenous isolation) or by host used to isolate the plasmid (exogenous isolation), it is not surprising that we have a poor appreciation of the diversity of the MGEs in many ecosystems. With this in mind we set out to develop an alternative approach to isolate plasmids from an ecosystem, in this case the human gut, which circumvented the limitations of current methods. Our approach, Transposon Aided Capture (TRACA), was successfully used to isolate plasmids from the distal portion of the human large intestine and we will discus the method and some of the TRACA plasmids in this presentation.

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