Characterization and genomic analysis of Sharanji: a jumbo bacteriophage of Escherichia coli

Abstract Background Bacteriophages are the most genetically diverse biological entities in nature. Our current understanding of phage biology primarily stems from studies on a limited number of model bacteriophages. Jumbo phages, characterized by their exceptionally large genomes, are less frequentl...

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Main Authors: Sharayu Magar, Sivaraj Barath, Debmitra Sen, Ranjith Kumar Singari, T. Nagarajan, Anjali Parmar, Sutharsan Govindarajan
Format: Article
Language:English
Published: BMC 2025-03-01
Series:Virology Journal
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Online Access:https://doi.org/10.1186/s12985-025-02646-5
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author Sharayu Magar
Sivaraj Barath
Debmitra Sen
Ranjith Kumar Singari
T. Nagarajan
Anjali Parmar
Sutharsan Govindarajan
author_facet Sharayu Magar
Sivaraj Barath
Debmitra Sen
Ranjith Kumar Singari
T. Nagarajan
Anjali Parmar
Sutharsan Govindarajan
author_sort Sharayu Magar
collection DOAJ
description Abstract Background Bacteriophages are the most genetically diverse biological entities in nature. Our current understanding of phage biology primarily stems from studies on a limited number of model bacteriophages. Jumbo phages, characterized by their exceptionally large genomes, are less frequently isolated and studied. Some jumbo phages exhibit remarkable genetic diversity, unique infection mechanisms, and therapeutic potential. Methods In this study, we describe the isolation of Sharanji, a novel Escherichia coli jumbo phage, isolated from chicken feces. The phage genome was sequenced and analyzed extensively through gene annotation and phylogenetic analysis. The jumbo phage was phenotypically characterized through electron microscopy, host range analysis, and survival at different pH and temperatures, and one-step growth curve assay. Finally, Sharanji mediated infection of E. coli is studied through fluorescence microscopy, to analyze its mechanism of infection compared to well-studied nucleus-forming jumbo phages. Results Whole genome sequencing reveals that Sharanji has a genome size of 350,079 bp and is a phage encompassing 593 ORFs. Genomic analysis indicates that the phage belongs to the Asteriusvirus genus and is related to E. coli jumbo phages PBECO4 and 121Q. Phenotypic analysis of isolated phage Sharanji, indicates that the phage size is 245.3 nm, and it is a narrow-spectrum phage infecting E. coli K12 strains, but not other bacteria including avian pathogenic E. coli. Infection analysis using microscopy shows that Sharanji infection causes cell filamentation. Furthermore, intracellular phage nucleus-like structures were not observed in Sharanji-infected cells, in contrast to infection by ΦKZ-like jumbo phages. Conclusions Our study reports the isolation and characterization of Sharanji, one of the large E. coli jumbo phages. Both genotypic and phenotypic analyses suggest that Sharanji serves as a unique model system for studying phage-bacteria interactions, particularly within the context of non-nucleus-forming jumbo phages. Further exploration of jumbo phages holds promise for uncovering new paradigms in the study of microbial viruses.
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spelling doaj-art-c6ade7a7d6cc4977991456bade7c011a2025-08-20T03:02:18ZengBMCVirology Journal1743-422X2025-03-0122111410.1186/s12985-025-02646-5Characterization and genomic analysis of Sharanji: a jumbo bacteriophage of Escherichia coliSharayu Magar0Sivaraj Barath1Debmitra Sen2Ranjith Kumar Singari3T. Nagarajan4Anjali Parmar5Sutharsan Govindarajan6Department of Biological Sciences, SRM University – APDepartment of Biological Sciences, SRM University – APDepartment of Biological Sciences, SRM University – APDepartment of Biological Sciences, SRM University – APDepartment of Biological Sciences, SRM University – APDepartment of Biological Sciences, SRM University – APDepartment of Biological Sciences, SRM University – APAbstract Background Bacteriophages are the most genetically diverse biological entities in nature. Our current understanding of phage biology primarily stems from studies on a limited number of model bacteriophages. Jumbo phages, characterized by their exceptionally large genomes, are less frequently isolated and studied. Some jumbo phages exhibit remarkable genetic diversity, unique infection mechanisms, and therapeutic potential. Methods In this study, we describe the isolation of Sharanji, a novel Escherichia coli jumbo phage, isolated from chicken feces. The phage genome was sequenced and analyzed extensively through gene annotation and phylogenetic analysis. The jumbo phage was phenotypically characterized through electron microscopy, host range analysis, and survival at different pH and temperatures, and one-step growth curve assay. Finally, Sharanji mediated infection of E. coli is studied through fluorescence microscopy, to analyze its mechanism of infection compared to well-studied nucleus-forming jumbo phages. Results Whole genome sequencing reveals that Sharanji has a genome size of 350,079 bp and is a phage encompassing 593 ORFs. Genomic analysis indicates that the phage belongs to the Asteriusvirus genus and is related to E. coli jumbo phages PBECO4 and 121Q. Phenotypic analysis of isolated phage Sharanji, indicates that the phage size is 245.3 nm, and it is a narrow-spectrum phage infecting E. coli K12 strains, but not other bacteria including avian pathogenic E. coli. Infection analysis using microscopy shows that Sharanji infection causes cell filamentation. Furthermore, intracellular phage nucleus-like structures were not observed in Sharanji-infected cells, in contrast to infection by ΦKZ-like jumbo phages. Conclusions Our study reports the isolation and characterization of Sharanji, one of the large E. coli jumbo phages. Both genotypic and phenotypic analyses suggest that Sharanji serves as a unique model system for studying phage-bacteria interactions, particularly within the context of non-nucleus-forming jumbo phages. Further exploration of jumbo phages holds promise for uncovering new paradigms in the study of microbial viruses.https://doi.org/10.1186/s12985-025-02646-5Jumbo phageE. coliBacteriophage genomics
spellingShingle Sharayu Magar
Sivaraj Barath
Debmitra Sen
Ranjith Kumar Singari
T. Nagarajan
Anjali Parmar
Sutharsan Govindarajan
Characterization and genomic analysis of Sharanji: a jumbo bacteriophage of Escherichia coli
Virology Journal
Jumbo phage
E. coli
Bacteriophage genomics
title Characterization and genomic analysis of Sharanji: a jumbo bacteriophage of Escherichia coli
title_full Characterization and genomic analysis of Sharanji: a jumbo bacteriophage of Escherichia coli
title_fullStr Characterization and genomic analysis of Sharanji: a jumbo bacteriophage of Escherichia coli
title_full_unstemmed Characterization and genomic analysis of Sharanji: a jumbo bacteriophage of Escherichia coli
title_short Characterization and genomic analysis of Sharanji: a jumbo bacteriophage of Escherichia coli
title_sort characterization and genomic analysis of sharanji a jumbo bacteriophage of escherichia coli
topic Jumbo phage
E. coli
Bacteriophage genomics
url https://doi.org/10.1186/s12985-025-02646-5
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