Characterization and Genomic Analysis of Phage asccφ28, a Phage of the Family Podoviridae Infecting Lactococcus lactis

Author:

Kotsonis Steven E.1,Powell Ian B.2,Pillidge Christopher J.2,Limsowtin Gaëtan K. Y.2,Hillier Alan J.3,Davidson Barrie E.1

Affiliation:

1. Department of Biochemistry and Molecular Biology, The University of Melbourne, Melbourne, Victoria 3010, Australia

2. Cultures Division, Dairy Innovation Australia, Ltd., Private Bag 16, Werribee, Victoria 3030, Australia

3. Food Science Australia, Private Bag 16, Werribee, Victoria 3030, Australia

Abstract

ABSTRACT Bacteriophage asccφ28 infects dairy fermentation strains of Lactococcus lactis . This report describes characterization of asccφ28 and its full genome sequence. Phage asccφ28 has a prolate head, whiskers, and a short tail (C2 morphotype). This morphology and DNA hybridization to L. lactis phage P369 DNA showed that asccφ28 belongs to the P034 phage species, a group rarely encountered in the dairy industry. The burst size of asccφ28 was found to be 121 ± 18 PFU per infected bacterial cell after a latent period of 44 min. The linear genome (18,762 bp) contains 28 possible open reading frames (ORFs) comprising 90% of the total genome. The ORFs are arranged bidirectionally in recognizable functional modules. The genome contains 577 bp inverted terminal repeats (ITRs) and putatively eight promoters and four terminators. The presence of ITRs, a phage-encoded DNA polymerase, and a terminal protein that binds to the DNA, along with BLAST and morphology data, show that asccφ28 more closely resembles streptococcal phage Cp-1 and the φ29-like phages that infect Bacillus subtilis than it resembles common lactococcal phages. The sequence of this phage is the first published sequence of a P034 species phage genome.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Reference46 articles.

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2. Allison, G. E., and T. R. Klaenhammer. 1998. Phage resistance mechanisms in lactic acid bacteria. Int. Dairy J.8:207-226.

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4. Anderson, D. L., and B. E. Reilly. 1993. Morphogenesis of bacteriophage φ29, p. 859-867. In A. L. Sonenshien, J. A. Hoch, and R. Losick (ed.), Bacillus subtilis and other gram-positive bacteria: biochemistry, physiology, and molecular genetics. American Society for Microbiology, Washington, DC.

5. Botstein, D. 1980. A theory of modular evolution for bacteriophages. Ann. N. Y. Acad. Sci.354:484-491.

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