Morphological and Genetic Characterization of Eggerthella lenta Bacteriophage PMBT5

is a common member of the human gut microbiome. We here describe the isolation and characterization of a putative virulent bacteriophage having as host. The double-layer agar method for isolating phages was adapted to anaerobic conditions for isolating bacteriophage PMBT5 from sewage on a strictly a...

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Bibliographic Details
Published in:Viruses 2022-07, Vol.14 (8), p.1598
Main Authors: Sprotte, Sabrina, Rasmussen, Torben S, Cho, Gyu-Sung, Brinks, Erik, Lametsch, René, Neve, Horst, Vogensen, Finn K, Nielsen, Dennis S, Franz, Charles M A P
Format: Article
Language:English
Subjects:
Actinobacteria
Agar
Amino acid sequence
anaerobe
Anaerobic conditions
Bacteria
Bacteriophages - genetics
Centrifugation
DNA, Viral - chemistry
DNA, Viral - genetics
Eggerthella lenta
Feces
genome sequence
Genome, Viral
Genomes
Humans
Intestinal microflora
Isometric
Metabolism
Microbiomes
Nitrogen
Phages
Physical characteristics
Pore size
Propagation
Sewage
Siphoviridae
Siphoviridae - genetics
Transmission electron microscopy
virulent phage
Work stations
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Summary:is a common member of the human gut microbiome. We here describe the isolation and characterization of a putative virulent bacteriophage having as host. The double-layer agar method for isolating phages was adapted to anaerobic conditions for isolating bacteriophage PMBT5 from sewage on a strictly anaerobic strain of intestinal origin. For this, anaerobically grown cells were concentrated by centrifugation and used for a 24 h phage enrichment step. Subsequently, this suspension was added to anaerobically prepared top (soft) agar in Hungate tubes and further used in the double-layer agar method. Based on morphological characteristics observed by transmission electron microscopy, phage PMBT5 could be assigned to the phage family. It showed an isometric head with a flexible, noncontractile tail and a distinct single 45 nm tail fiber under the baseplate. Genome sequencing and assembly resulted in one contig of 30,930 bp and a mol% GC content of 51.3, consisting of 44 predicted protein-encoding genes. Phage-related proteins could be largely identified based on their amino acid sequence, and a comparison with metagenomes in the human virome database showed that the phage genome exhibits similarity to two distantly related phages.
ISSN:1999-4915
1999-4915
DOI:10.3390/v14081598