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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 ascc{phi}28, a Phage of the Family Podoviridae Infecting Lactococcus lactis{triangledown}

Steven E. Kotsonis,1,{dagger} 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 ascc{phi}28 infects dairy fermentation strains of Lactococcus lactis. This report describes characterization of ascc{phi}28 and its full genome sequence. Phage ascc{phi}28 has a prolate head, whiskers, and a short tail (C2 morphotype). This morphology and DNA hybridization to L. lactis phage P369 DNA showed that ascc{phi}28 belongs to the P034 phage species, a group rarely encountered in the dairy industry. The burst size of ascc{phi}28 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 ascc{phi}28 more closely resembles streptococcal phage Cp-1 and the {phi}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.

* Corresponding author. Mailing address: Cultures Division, Dairy Innovation Australia, Ltd., Private Bag 16, Werribee, Victoria 3030, Australia. Phone: 61 (3) 9974 8910. Fax: 61 (3) 9742 4233. E-mail: ipowell{at}dairyinnovation.com.au

{triangledown} Published ahead of print on 4 April 2008.

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

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.





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