Morphological and Genetic Characterization of Eggerthella lenta Bacteriophage PMBT5.
Sabrina KobergTorben Sølbeck RasmussenGyu-Sung ChoErik BrinksRené LametschHorst NeveFinn K VogensenDennis Sandris NielsenCharles M A P FranzPublished in: Viruses (2022)
Eggerthella lenta is a common member of the human gut microbiome. We here describe the isolation and characterization of a putative virulent bacteriophage having E. lenta 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 E. lenta strain of intestinal origin. For this, anaerobically grown E. lenta 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 Siphoviridae 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.
Keyphrases
- pseudomonas aeruginosa
- amino acid
- endothelial cells
- microbial community
- genome wide
- wastewater treatment
- electron microscopy
- induced pluripotent stem cells
- induced apoptosis
- pluripotent stem cells
- cell cycle arrest
- cell death
- photodynamic therapy
- gene expression
- emergency department
- risk assessment
- dna methylation
- high resolution
- sewage sludge
- endoplasmic reticulum stress
- transcription factor
- antibiotic resistance genes
- protein protein
- solid phase extraction
- simultaneous determination