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Open Access Research article

The Bacillus anthracis chromosome contains four conserved, excision-proficient, putative prophages

Shanmuga Sozhamannan1*, Michael D Chute1, Farrell D McAfee1, Derrick E Fouts2, Arya Akmal1, Darrell R Galloway13, Alfred Mateczun1, Leslie W Baillie13 and Timothy D Read13

Author affiliations

1 Biological Defense Research Directorate, Naval Medical Research Center, 503 Robert Grant Avenue, Silver Spring, Maryland 20852, USA

2 The Institute for Genomic Research, Rockville, Maryland 20850, USA

3 University of Maryland Medical Biotechnology Center, Baltimore, Maryland 21201, USA

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Citation and License

BMC Microbiology 2006, 6:34  doi:10.1186/1471-2180-6-34

Published: 6 April 2006

Abstract

Background

Bacillus anthracis is considered to be a recently emerged clone within the Bacillus cereus sensu lato group. The B. anthracis genome sequence contains four putative lambdoid prophages. We undertook this study in order to understand whether the four prophages are unique to B. anthracis and whether they produce active phages.

Results

More than 300 geographically and temporally divergent isolates of B. anthracis and its near neighbors were screened by PCR for the presence of specific DNA sequences from each prophage region. Every isolate of B. anthracis screened by PCR was found to produce all four phage-specific amplicons whereas none of the non-B. anthracis isolates, produced more than one phage-specific amplicon. Excision of prophages could be detected by a PCR based assay for attP sites on extra-chromosomal phage circles and for attB sites on phage-excised chromosomes. SYBR-green real-time PCR assays indicated that prophage excision occurs at very low frequencies (2 × 10-5 - 8 × 10-8/cell). Induction with mitomycin C increased the frequency of excision of one of the prophages by approximately 250 fold. All four prophages appear to be defective since, mitomycin C induced culture did not release any viable phage particle or lyse the cells or reveal any phage particle under electron microscopic examination.

Conclusion

The retention of all four putative prophage regions across all tested strains of B. anthracis is further evidence of the very recent emergence of this lineage and the prophage regions may be useful for differentiating the B. anthracis chromosome from that of its neighbors. All four prophages can excise at low frequencies, but are apparently defective in phage production.