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Journal of Bacteriology, July 2002, p. 4003-4017, Vol. 184, No. 14
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.14.4003-4017.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Burkholderia thailandensis E125 Harbors a Temperate Bacteriophage Specific for Burkholderia mallei

Donald E. Woods,1 Jeffrey A. Jeddeloh,2 David L. Fritz,3 and David DeShazer2*

Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, Calgary, Alberta T2N 4N1, Canada,1 Bacteriology,2 Pathology Divisions, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland 217023

Received 5 February 2002/ Accepted 12 April 2002

Burkholderia thailandensis is a nonpathogenic gram-negative bacillus that is closely related to Burkholderia mallei and Burkholderia pseudomallei. We found that B. thailandensis E125 spontaneously produced a bacteriophage, termed {phi}E125, which formed turbid plaques in top agar containing B. mallei ATCC 23344. We examined the host range of {phi}E125 and found that it formed plaques on B. mallei but not on any other bacterial species tested, including B. thailandensis and B. pseudomallei. Examination of the bacteriophage by transmission electron microscopy revealed an isometric head and a long noncontractile tail. B. mallei NCTC 120 and B. mallei DB110795 were resistant to infection with {phi}E125 and did not produce lipopolysaccharide (LPS) O antigen due to IS407A insertions in wbiE and wbiG, respectively. wbiE was provided in trans on a broad-host-range plasmid to B. mallei NCTC 120, and it restored LPS O-antigen production and susceptibility to {phi}E125. The 53,373-bp {phi}E125 genome contained 70 genes, an IS3 family insertion sequence (ISBt3), and an attachment site (attP) encompassing the 3' end of a proline tRNA (UGG) gene. While the overall genetic organization of the {phi}E125 genome was similar to {lambda}-like bacteriophages and prophages, it also possessed a novel cluster of putative replication and lysogeny genes. The {phi}E125 genome encoded an adenine and a cytosine methyltransferase, and purified bacteriophage DNA contained both N6-methyladenine and N4-methylcytosine. The results presented here demonstrate that {phi}E125 is a new member of the {lambda} supergroup of Siphoviridae that may be useful as a diagnostic tool for B. mallei.

* Corresponding author. Mailing address: 1425 Porter St., USAMRIID, Bacteriology Division, Fort Detrick, MD 21702. Phone: (301) 619-4871. Fax: (301) 619-4898. E-mail: david.deshazer{at}amedd.army.mil.

Journal of Bacteriology, July 2002, p. 4003-4017, Vol. 184, No. 14
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.14.4003-4017.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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