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Applied and Environmental Microbiology, August 2001, p. 3564-3576, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3564-3576.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Sequence Analysis and Molecular Characterization of the Lactococcus lactis Temperate Bacteriophage BK5-T†

Chitladda Mahanivong,1 John D. Boyce,1,Dagger Barrie E. Davidson,1 and Alan J. Hillier2,3,*

Russell Grimwade School of Biochemistry and Molecular Biology, The University of Melbourne, Parkville, Victoria 3010,1 and Food Science Australia2 and Department of Food Science and Agribusiness, The University of Melbourne,3 Werribee, Victoria 3030, Australia

Received 8 December 2000/Accepted 20 April 2001

The Lactococcus lactis temperate bacteriophage BK5-T is one of twelve type phages that define L. lactis phage species. This paper describes the nucleotide sequence and analysis of a 21-kbp region of the BK5-T genome and completes the nucleotide sequence of the genome of this phage. The 40,003-nucleotide linear genome encodes 63 open reading frames. Sequence runoff experiments showed that the cohesive ends of the BK5-T genome contained a 12-bp 3' single-stranded overhang with the sequence 5'-CACACACATAGG-3'. Two major BK5-T structural proteins, of approximately 30 and 20 kDa, were identified, and N-terminal sequence analysis determined that they were encoded by orf7 and orf12, respectively. A 169-bp fragment containing a 37-bp direct repeat and several smaller repeat sequences conferred resistance to BK5-T infection when introduced in trans to the host cell and is likely a part of the BK5-T origin of replication (ori).

* Corresponding author. Mailing address: Food Science Australia, Private Bag 16, Werribee, Victoria 3030, Australia. Phone: 61 3 9731 3268. Fax: 61 3 9731 3254. E-mail: alan.hillier{at}foodscience.afisc.csiro.au.

dagger This report is dedicated to the memory of Barrie E. Davidson, who passed away in July 2000.

Dagger Present address: Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia.

Applied and Environmental Microbiology, August 2001, p. 3564-3576, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3564-3576.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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