文章基本信息

标题：Isolation of Genetically Tractable Most-Wanted Bacteria by Metaparental Mating
本地全文：下载
作者：Páraic Ó Cuív ; Wendy J. Smith ; Sian Pottenger 等
期刊名称：Scientific Reports
电子版ISSN：2045-2322
出版年度：2015
卷号：5
DOI：10.1038/srep13282
出版社：Springer Nature
摘要：Metagenomics has rapidly advanced our inventory and appreciation of the genetic potential inherent to the gut microbiome. However it is widely accepted that two key constraints to further genetic dissection of the gut microbiota and host-microbe interactions have been our inability to recover new isolates from the human gut, and the paucity of genetically tractable gut microbes. To address this challenge we developed a modular RP4 mobilisable recombinant vector system and an approach termed metaparental mating to support the rapid and directed isolation of genetically tractable fastidious gut bacteria. Using this approach we isolated transconjugants affiliated with Clostridium cluster IV ( Faecalibacterium and Oscillibacter spp.), Clostridium cluster XI ( Anaerococcus ) and Clostridium XIVa ( Blautia spp.) and group 2 ruminococci amongst others, and demonstrated that the recombinant vectors were stably maintained in their recipient hosts. By a similar approach we constructed fluorescently labelled bacterial transconjugants affiliated with Clostridium cluster IV (including Flavonifractor and Pseudoflavonifractor spp.), Clostridium XIVa ( Blautia spp.) and Clostridium cluster XVIII ( Clostridium ramosum ) that expressed a flavin mononucleotide-based reporter gene ( evoglow-C-Bs2 ). Our approach will advance the integration of bacterial genetics with metagenomics and realize new directions to support a more mechanistic dissection of host-microbe associations relevant to human health and disease.