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

mBio

Panagiota Stamou¹  Tadhg Crowley¹  Kieran James¹  Ken Nally¹  Marco Ventura²  Gabriele A. Lugli²  Silvia Arboleya³  Miguel Gueimonde³  Fabio dal Bello⁴  Ines Martinez⁴  Ian O’Neill⁵  Katherine Lavelle⁵  Kelsey White⁵  Douwe van Sinderen⁵  Stephen Hayes⁵  Jennifer Mahony⁵  Brian McDonnell⁵  Eoghan Casey⁵ 

[1]APC Microbiome Ireland, University College Cork, Cork, Ireland

[2]Laboratory of Probiogenomics, Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, Parma, Italy

[3]Microbiota, Food and Health Group, Department of Biochemistry and Microbiology of Dairy Products, Institutode Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas, Villaviciosa, Spain

[4]Sacco Srl, Cadorago (Co), Italy

[5]School of Microbiology, University College Cork, Cork, Ireland

关键词: metagenome;    virome;    phageome;    cytometry;    fluorescent;    RBP;   

DOI  :  10.1128/mbio.03334-21

来源: DOAJ

【 摘 要 】

ABSTRACT 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.

【 授权许可】

Unknown