Characterization and Genome Sequence of Marine Alteromonas gracilis Phage PB15 Isolated from the Yellow Sea, China

Gao, Yu; Liu, Qian; Wang, Min; Zhao, Guihua; Malin, Gill; Gong, Zheng; Meng, Xue; Lin, Tongtong; Li, Yutong; Shao, Hongbing

doi:10.1007/s00284-017-1251-9

Cited by 17 publications

(22 citation statements)

References 26 publications

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“…The first Alteromonas phages characterized were isolated from western Mediterranean coastal waters and identified as a new genus within the Podoviridae family (17). An Alteromonas-targeting Podoviridae phage isolated from the North Sea (18) has been used as a biological tracer in hydrological transport studies (19), and finally two Alteromonas phages belonging to the Siphoviridae family have been isolated from the western Yellow Sea (20,21). These (seven in total) are the only Alteromonas phages isolated and characterized to date.…”

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confidence: 99%

Alteromonas Myovirus V22 Represents a New Genus of Marine Bacteriophages Requiring a Tail Fiber Chaperone for Host Recognition

et al. 2020

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Marine phages play a variety of critical roles in regulating the microbial composition of our oceans. Despite constituting the majority of genetic diversity within these environments, there are relatively few isolates with complete genome sequences or in-depth analyses of their host interaction mechanisms, such as characterization of their receptor binding proteins (RBPs). Here, we present the 92,760-bp genome of the Alteromonas-targeting phage V22. Genomic and morphological analyses identify V22 as a myovirus; however, due to a lack of sequence similarity to any other known myoviruses, we propose that V22 be classified as the type phage of a new Myoalterovirus genus within the Myoviridae family. V22 shows gene homology and synteny with two different subfamilies of phages infecting enterobacteria, specifically within the structural region of its genome. To improve our understanding of the V22 adsorption process, we identified putative RBPs (gp23, gp24, and gp26) and tested their ability to decorate the V22 propagation strain, Alteromonas mediterranea PT11, as recombinant green fluorescent protein (GFP)-tagged constructs. Only GFP-gp26 was capable of bacterial recognition and identified as the V22 RBP. Interestingly, production of functional GFP-gp26 required coexpression with the downstream protein gp27. GFP-gp26 could be expressed alone but was incapable of host recognition. By combining size-exclusion chromatography with fluorescence microscopy, we reveal how gp27 is not a component of the final RBP complex but instead is identified as a new type of phage-encoded intermolecular chaperone that is essential for maturation of the gp26 RBP. IMPORTANCE Host recognition by phage-encoded receptor binding proteins (RBPs) constitutes the first step in all phage infections and the most critical determinant of host specificity. By characterizing new types of RBPs and identifying their essential chaperones, we hope to expand the repertoire of known phage-host recognition machineries. Due to their genetic plasticity, studying RBPs and their associated chaperones can shed new light onto viral evolution affecting phage-host interactions, which is essential for fields such as phage therapy or biotechnology. In addition, since marine phages constitute one of the most important reservoirs of noncharacterized genetic diversity on the planet, their genomic and functional characterization may be of paramount importance for the discovery of novel genes with potential applications.

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Alteromonas Myovirus V22 Represents a New Genus of Marine Bacteriophages Requiring a Tail Fiber Chaperone for Host Recognition

et al. 2020

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“…The burst size is an important criterion that should be estimated to understand the replication dynamics of the isolated phage [39]. It was reported that burst sizes are variable in Siphovirus (between 60 and 147 PFU/mL), whereas latent periods and rise times range from 16 to 55 min and 24 to 65 min, respectively [40,41]. Phages have different activities against bacteria, including (i) bacteriostatic activity that stops the bacterial growth, as in Acinetobacter baumannii after 360 min of incubation with phages [42,43], and P. aeruginosa [44]; and (ii) bacteriolytic activity, represented by the reduction in bacterial OD 600 from starting concentration, as in A. baumannii [45] and Aeromonas hydrophila [46].…”

Section: Discussionmentioning

confidence: 99%

“…To investigate the antibacterial effect of phage ZCSE6 as described in [24], host bacterial cultures at an optical density at 600 nm (OD 600 ) of 0.4, which is equivalent to 10 8 CFU/mL, were infected with phage at different MOIs (0.1, 0.01, and 0.001) and incubated at 37 • C for 7 h. During the incubation time, the absorbance changes at 600 nm (OD 600 ) were measured using Jenway 7200 visible spectrophotometer (Jenway, Staffordshire, UK) at various time points (0, 15,40,60,75,90,120,165,195,225,360,390, and 420 min).…”

Section: Time-killing Curvesmentioning

confidence: 99%

Isolation and Characterization of Bacteriophage ZCSE6 against Salmonella spp.: Phage Application in Milk

et al. 2021

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Food safety is very important in the food industry as most pathogenic bacteria can cause food-borne diseases and negatively affect public health. In the milk industry, contamination with Salmonella has always been a challenge, but the risks have dramatically increased as almost all bacteria now show resistance to a wide range of commercial antibiotics. This study aimed to isolate a bacteriophage to be used as a bactericidal agent against Salmonella in milk and dairy products. Here, phage ZCSE6 has been isolated from raw milk sample sand molecularly and chemically characterized. At different multiplicities of infection (MOIs) of 0.1, 0.01, and 0.001, the phage–Salmonella interaction was studied for 6 h at 37 °C and 24 h at 8 °C. In addition, ZCSE6 was tested against Salmonella contamination in milk to examine its lytic activity for 3 h at 37 °C. The results showed that ZCSE6 has a small genome size (<48.5 kbp) and belongs to the Siphovirus family. Phage ZCSE6 revealed a high thermal and pH stability at various conditions that mimic milk manufacturing and supply chain conditions. It also demonstrated a significant reduction in Salmonella concentration in media at various MOIs, with higher bacterial eradication at higher MOI. Moreover, it significantly reduced Salmonella growth (MOI 1) in milk, manifesting a 1000-fold decrease in bacteria concentration following 3 h incubation at 37 °C. The results highlighted the strong ability of ZCSE6 to kill Salmonella and control its growth in milk. Thus, ZCSE6 is recommended as a biocontrol agent in milk to limit bacterial growth and increase the milk shelf-life.

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“…Stained particles were observed using a JEOL Model JEM-1200EX TEM at 100 kV. Phage dimensions were estimated based on the electron micrographs ( Gao et al, 2017 ).…”

Section: Methodsmentioning

confidence: 99%

Characterization and Genomic Analysis of Marinobacter Phage vB_MalS-PS3, Representing a New Lambda-Like Temperate Siphoviral Genus Infecting Algae-Associated Bacteria

et al. 2021

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Marinobacter is the abundant and important algal-associated and hydrocarbon biodegradation bacteria in the ocean. However, little knowledge about their phages has been reported. Here, a novel siphovirus, vB_MalS-PS3, infecting Marinobacter algicola DG893(T), was isolated from the surface waters of the western Pacific Ocean. Transmission electron microscopy (TEM) indicated that vB_MalS-PS3 has the morphology of siphoviruses. VB_MalS-PS3 was stable from −20 to 55°C, and with the latent and rise periods of about 80 and 10 min, respectively. The genome sequence of VB_MalS-PS3 contains a linear, double-strand 42,168-bp DNA molecule with a G + C content of 56.23% and 54 putative open reading frames (ORFs). Nineteen conserved domains were predicted by BLASTp in NCBI. We found that vB_MalS-PS3 represent an understudied viral group with only one known isolate. The phylogenetic tree based on the amino acid sequences of whole genomes revealed that vB_MalS-PS3 has a distant evolutionary relationship with other siphoviruses, and can be grouped into a novel viral genus cluster with six uncultured assembled viral genomes from metagenomics, named here as Marinovirus. This study of the Marinobacter phage vB_MalS-PS3 genome enriched the genetic database of marine bacteriophages, in addition, will provide useful information for further research on the interaction between Marinobacter phages and their hosts, and their relationship with algal blooms and hydrocarbon biodegradation in the ocean.

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Characterization and Genome Sequence of Marine Alteromonas gracilis Phage PB15 Isolated from the Yellow Sea, China

Cited by 17 publications

References 26 publications

Alteromonas Myovirus V22 Represents a New Genus of Marine Bacteriophages Requiring a Tail Fiber Chaperone for Host Recognition

Alteromonas Myovirus V22 Represents a New Genus of Marine Bacteriophages Requiring a Tail Fiber Chaperone for Host Recognition

Isolation and Characterization of Bacteriophage ZCSE6 against Salmonella spp.: Phage Application in Milk

Characterization and Genomic Analysis of Marinobacter Phage vB_MalS-PS3, Representing a New Lambda-Like Temperate Siphoviral Genus Infecting Algae-Associated Bacteria

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