Morphological and Genetic Characterization of Eggerthella lenta Bacteriophage PMBT5
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Morphological and Genetic Characterization of Eggerthella lenta Bacteriophage PMBT5. / 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.
In: Viruses, Vol. 14, No. 8, 1598, 2022.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Morphological and Genetic Characterization of Eggerthella lenta Bacteriophage PMBT5
AU - Sprotte, Sabrina
AU - Rasmussen, Torben S.
AU - Cho, Gyu-Sung
AU - Brinks, Erik
AU - Lametsch, René
AU - Neve, Horst
AU - Vogensen, Finn K.
AU - Nielsen, Dennis S.
AU - Franz, Charles M.A.P.
N1 - Publisher Copyright: © 2022 by the authors.
PY - 2022
Y1 - 2022
N2 - 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.
AB - 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.
KW - anaerobe
KW - Eggerthella lenta
KW - genome sequence
KW - Siphoviridae
KW - virulent phage
U2 - 10.3390/v14081598
DO - 10.3390/v14081598
M3 - Journal article
C2 - 35893664
AN - SCOPUS:85137407365
VL - 14
JO - Viruses
JF - Viruses
SN - 1999-4915
IS - 8
M1 - 1598
ER -
ID: 319602930