4.7 Article

Needle in a Whey-Stack: PhRACS as a Discovery Tool for Unknown Phage-Host Combinations

期刊

MBIO
卷 13, 期 1, 页码 -

出版社

AMER SOC MICROBIOLOGY
DOI: 10.1128/mbio.03334-21

关键词

metagenome; virome; phageome; cytometry; fluorescent; RBP; fecal; whey; fluorescence

资金

  1. EU Joint Programming Initiative -A Healthy Diet for a Healthy Life
  2. Science Foundation Ireland [SFI] [15/JP-HDHL/3280]
  3. (MIUR, Italy) - SFI through the Irish Government's National Development Plan [SFI/12/RC/2273a, SFI/12/RC/2273b]
  4. SFI-funded Starting Investigator Research Grant (SIRG) [15/SIRG/3430]
  5. postdoctoral Juan de la Cierva Contract (Ministry of Science, Innovation and Universities) [IJCI-2017-32156]
  6. Science Foundation Ireland [15/JP-HDHL/3280, SFI/12/RC/2273a, SFI/12/RC/2273b, 15/SIRG/3430]
  7. Science Foundation Ireland (SFI) [15/JP-HDHL/3280, 15/SIRG/3430] Funding Source: Science Foundation Ireland (SFI)

向作者/读者索取更多资源

This study describes a method that exploits the interaction between specific phage receptor binding proteins (RBPs) and host cell surface receptors to isolate phage-host combinations from environmental samples.
The field of metagenomics has rapidly expanded to become the go-to method for complex microbial community analyses. However, there is currently no straightforward route from metagenomics to traditional culture-based methods of strain isolation, particularly in (bacterio)phage biology, leading to an investigative bottleneck. Here, we describe a method that exploits specific phage receptor binding protein (RBP)-host cell surface receptor interaction enabling isolation of phage-host combinations from an environmental sample. The method was successfully applied to two complex sample types-a dairy-derived whey sample and an infant fecal sample, enabling retrieval of specific and culturable phage hosts. IMPORTANCE PhRACS aims to bridge the current divide between in silico genetic analyses (i.e., phageomic studies) and traditional culture-based methodology. Through the labeling of specific bacterial hosts with fluorescently tagged recombinant phage receptor binding proteins and the isolation of tagged cells using flow cytometry, PhRACS allows the full potential of phageomic data to be realized in the wet laboratory.

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