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Characterization of a new phage, termed ϕA318, which is specific for Vibrio alginolyticus

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Abstract

Vibrio alginolyticus is an opportunistic pathogen of animals and humans; its related strains can also produce tetrodotoxin and hemolysins. A new phage, ϕA318, which lysed its host V. alginolyticus with high efficiency, was characterized. The burst size of ϕA318 in V. alginolyticus was 72 PFU/bacterium at an MOI of 1 at room temperature; the plaque size was as large as 5 mm in diameter. Electron microscopy (EM) of the phage particles revealed a 50- to 55-nm isomorphous icosahedral head with a 12-nm non-contractile tail, similar to the T7-like phages of the family Podoviridae. Phylogenetic analysis based on complete sequences of the DNA-directed RNA polymerase gene revealed that ϕA318 had 28-47% amino acid identity to enterobacteria phages T7 and SP6, and other Vibrio phages, and the phylogenetic distance suggested that ϕA318 could be classified as a new T7-like bacteriophage. Nevertheless, several motifs in the ϕA318 phage RNA polymerase were highly conserved, including DFRGR (T7-421 motif), DG (T7-537 motif), PSEKPQDIYGAVS (T7-563 motif), RSMTKKPVMTL PYGS (T7-627 motif), and HDS (T7-811 motif). Genetic analysis indicated that phage ϕA318 is not a thermostable direct hemolysin producer. The results suggest that the MOI should be higher than 0.1 to prevent the chance of hemolysin production by the bacteria before they are lysed by the phage.

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Acknowledgments

This research fund is partially supported by grants from the National Science Council, Taiwan (NSC96-2313-B-110-002-MY3 and NSC99-2313-B-110-002-MY3), and the Ministry of Education, Taiwan (NSYSU95-99C031701; the second term of Top University Program: NSYSU 00C030205 and NCHU 100-S05-09) under the ATU plan. We thank Professor Long-Huw Lee (National Chung-Hsing University) as the grant organizer of the intercampus ATU plan. Also we appreciate Yu-Tin Liu for helping with gel preparation, and Kenneth B. Lin and Dr. Simon White for comments and editing.

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  1. Department of Marine Biotechnology and Resources, Asia-Pacific Ocean Research Center, National Sun Yat-sen University, Kaohsiung, 80424, Taiwan

    Ying-Rong Lin, Chi-Wen Chiu, Feng-Yi Chang & Chan-Shing Lin

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  1. Ying-Rong Lin
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  2. Chi-Wen Chiu
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Correspondence to Chan-Shing Lin.

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Lin, YR., Chiu, CW., Chang, FY. et al. Characterization of a new phage, termed ϕA318, which is specific for Vibrio alginolyticus . Arch Virol 157, 917–926 (2012). https://doi.org/10.1007/s00705-012-1244-8

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