Isolation and engineering of a Listeria grayi bacteriophage

Abstract The lack of bacteriophages capable of infecting the Listeria species, Listeria grayi, is academically intriguing and presents an obstacle to the development of bacteriophage-based technologies for Listeria. We describe the isolation and engineering of a novel L. grayi bacteriophage, LPJP1,...

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Autores principales:	Stephen Erickson, John Paulson, Matthew Brown, Wendy Hahn, Jose Gil, Rocío Barron-Montenegro, Andrea I. Moreno-Switt, Marcia Eisenberg, Minh M. Nguyen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/58a93fdc7cb4498ba580acc31f890973
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spelling	oai:doaj.org-article:58a93fdc7cb4498ba580acc31f8909732021-12-02T15:15:13ZIsolation and engineering of a Listeria grayi bacteriophage10.1038/s41598-021-98134-12045-2322https://doaj.org/article/58a93fdc7cb4498ba580acc31f8909732021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-98134-1https://doaj.org/toc/2045-2322Abstract The lack of bacteriophages capable of infecting the Listeria species, Listeria grayi, is academically intriguing and presents an obstacle to the development of bacteriophage-based technologies for Listeria. We describe the isolation and engineering of a novel L. grayi bacteriophage, LPJP1, isolated from farm silage. With a genome over 200,000 base pairs, LPJP1 is the first and only reported jumbo bacteriophage infecting the Listeria genus. Similar to other Gram-positive jumbo phages, LPJP1 appeared to contain modified base pairs, which complicated initial attempts to obtain genomic sequence using standard methods. Following successful sequencing with a modified approach, a recombinant of LPJP1 encoding the NanoLuc luciferase was engineered using homologous recombination. This luciferase reporter bacteriophage successfully detected 100 stationary phase colony forming units of both subspecies of L. grayi in four hours. A single log phase colony forming unit was also sufficient for positive detection in the same time period. The recombinant demonstrated complete specificity for this particular Listeria species and did not infect 150 non-L. grayi Listeria strains nor any other bacterial genus. LPJP1 is believed to be the first reported lytic bacteriophage of L. grayi as well as the only jumbo bacteriophage to be successfully engineered into a luciferase reporter.Stephen EricksonJohn PaulsonMatthew BrownWendy HahnJose GilRocío Barron-MontenegroAndrea I. Moreno-SwittMarcia EisenbergMinh M. NguyenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Stephen Erickson John Paulson Matthew Brown Wendy Hahn Jose Gil Rocío Barron-Montenegro Andrea I. Moreno-Switt Marcia Eisenberg Minh M. Nguyen Isolation and engineering of a Listeria grayi bacteriophage
description	Abstract The lack of bacteriophages capable of infecting the Listeria species, Listeria grayi, is academically intriguing and presents an obstacle to the development of bacteriophage-based technologies for Listeria. We describe the isolation and engineering of a novel L. grayi bacteriophage, LPJP1, isolated from farm silage. With a genome over 200,000 base pairs, LPJP1 is the first and only reported jumbo bacteriophage infecting the Listeria genus. Similar to other Gram-positive jumbo phages, LPJP1 appeared to contain modified base pairs, which complicated initial attempts to obtain genomic sequence using standard methods. Following successful sequencing with a modified approach, a recombinant of LPJP1 encoding the NanoLuc luciferase was engineered using homologous recombination. This luciferase reporter bacteriophage successfully detected 100 stationary phase colony forming units of both subspecies of L. grayi in four hours. A single log phase colony forming unit was also sufficient for positive detection in the same time period. The recombinant demonstrated complete specificity for this particular Listeria species and did not infect 150 non-L. grayi Listeria strains nor any other bacterial genus. LPJP1 is believed to be the first reported lytic bacteriophage of L. grayi as well as the only jumbo bacteriophage to be successfully engineered into a luciferase reporter.
format	article
author	Stephen Erickson John Paulson Matthew Brown Wendy Hahn Jose Gil Rocío Barron-Montenegro Andrea I. Moreno-Switt Marcia Eisenberg Minh M. Nguyen
author_facet	Stephen Erickson John Paulson Matthew Brown Wendy Hahn Jose Gil Rocío Barron-Montenegro Andrea I. Moreno-Switt Marcia Eisenberg Minh M. Nguyen
author_sort	Stephen Erickson
title	Isolation and engineering of a Listeria grayi bacteriophage
title_short	Isolation and engineering of a Listeria grayi bacteriophage
title_full	Isolation and engineering of a Listeria grayi bacteriophage
title_fullStr	Isolation and engineering of a Listeria grayi bacteriophage
title_full_unstemmed	Isolation and engineering of a Listeria grayi bacteriophage
title_sort	isolation and engineering of a listeria grayi bacteriophage
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/58a93fdc7cb4498ba580acc31f890973
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Isolation and engineering of a Listeria grayi bacteriophage

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