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Applied and Environmental Microbiology, June 2008, p. 3453-3460, Vol. 74, No. 11
0099-2240/08/$08.00+0     doi:10.1128/AEM.02379-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Characterization and Genomic Analysis of Phage ascc28, a Phage of the Family Podoviridae Infecting Lactococcus lactis

Steven E. Kotsonis,^1, Ian B. Powell,^2* Christopher J. Pillidge,² Gaëtan K. Y. Limsowtin,² Alan J. Hillier,³ and Barrie E. Davidson¹

Department of Biochemistry and Molecular Biology, The University of Melbourne, Melbourne, Victoria 3010, Australia,¹ Cultures Division, Dairy Innovation Australia, Ltd., Private Bag 16, Werribee, Victoria 3030, Australia,² Food Science Australia, Private Bag 16, Werribee, Victoria 3030, Australia³

Received 23 October 2007/ Accepted 23 March 2008

Bacteriophage ascc28 infects dairy fermentation strains of Lactococcus lactis. This report describes characterization of ascc28 and its full genome sequence. Phage ascc28 has a prolate head, whiskers, and a short tail (C2 morphotype). This morphology and DNA hybridization to L. lactis phage P369 DNA showed that ascc28 belongs to the P034 phage species, a group rarely encountered in the dairy industry. The burst size of ascc28 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 ascc28 more closely resembles streptococcal phage Cp-1 and the 29-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.

Published ahead of print on 4 April 2008.

Present address: Orica Pty Limited, 1 Nicholson Street, Melbourne, Victoria 3000, Australia.