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. 1997 Feb;179(3):818–824. doi: 10.1128/jb.179.3.818-824.1997

Molecular evolution and diversity in Bacillus anthracis as detected by amplified fragment length polymorphism markers.

P Keim 1, A Kalif 1, J Schupp 1, K Hill 1, S E Travis 1, K Richmond 1, D M Adair 1, M Hugh-Jones 1, C R Kuske 1, P Jackson 1
PMCID: PMC178765  PMID: 9006038

Abstract

Bacillus anthracis causes anthrax and represents one of the most molecularly monomorphic bacteria known. We have used AFLP (amplified fragment length polymorphism) DNA markers to analyze 78 B. anthracis isolates and six related Bacillus species for molecular variation. AFLP markers are extremely sensitive to even small sequence variation, using PCR and high-resolution electrophoresis to examine restriction fragments. Using this approach, we examined ca. 6.3% of the Bacillus genome for length mutations and ca. 0.36% for point mutations. Extensive variation was observed among taxa, and both cladistic and phenetic analyses were used to construct a phylogeny of B. anthracis and its closest relatives. This genome-wide analysis of 357 AFLP characters (polymorphic fragments) indicates that B. cereus and B. thuringiensis are the closest taxa to B. anthracis, with B. mycoides slightly more distant. B. subtilis, B. polymyxa, and B. stearothermophilus shared few AFLP markers with B. anthracis and were used as outgroups to root the analysis. In contrast to the variation among taxa, only rare AFLP marker variation was observed within B. anthracis, which may be the most genetically uniform bacterial species known. However, AFLP markers did establish the presence or absence of the pXO1 and pXO2 plasmids and detected 31 polymorphic chromosomal regions among the 79 B. anthracis isolates. Cluster analysis identified two very distinct genetic lineages among the B. anthracis isolates. The level of variation and its geographic distribution are consistent with a historically recent African origin for this pathogenic organism. Based on AFLP marker similarity, the ongoing anthrax epidemic in Canada and the northern United States is due to a single strain introduction that has remained stable over at least 30 years and a 1,000-mile distribution.

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Selected References

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  1. Andersen G. L., Simchock J. M., Wilson K. H. Identification of a region of genetic variability among Bacillus anthracis strains and related species. J Bacteriol. 1996 Jan;178(2):377–384. doi: 10.1128/jb.178.2.377-384.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ash C., Collins M. D. Comparative analysis of 23S ribosomal RNA gene sequences of Bacillus anthracis and emetic Bacillus cereus determined by PCR-direct sequencing. FEMS Microbiol Lett. 1992 Jul 1;73(1-2):75–80. doi: 10.1016/0378-1097(92)90586-d. [DOI] [PubMed] [Google Scholar]
  3. Ash C., Farrow J. A., Dorsch M., Stackebrandt E., Collins M. D. Comparative analysis of Bacillus anthracis, Bacillus cereus, and related species on the basis of reverse transcriptase sequencing of 16S rRNA. Int J Syst Bacteriol. 1991 Jul;41(3):343–346. doi: 10.1099/00207713-41-3-343. [DOI] [PubMed] [Google Scholar]
  4. Drobniewski F. A. Bacillus cereus and related species. Clin Microbiol Rev. 1993 Oct;6(4):324–338. doi: 10.1128/cmr.6.4.324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Groenen P. M., Bunschoten A. E., van Soolingen D., van Embden J. D. Nature of DNA polymorphism in the direct repeat cluster of Mycobacterium tuberculosis; application for strain differentiation by a novel typing method. Mol Microbiol. 1993 Dec;10(5):1057–1065. doi: 10.1111/j.1365-2958.1993.tb00976.x. [DOI] [PubMed] [Google Scholar]
  6. Harrell L. J., Andersen G. L., Wilson K. H. Genetic variability of Bacillus anthracis and related species. J Clin Microbiol. 1995 Jul;33(7):1847–1850. doi: 10.1128/jcm.33.7.1847-1850.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Henderson I., Duggleby C. J., Turnbull P. C. Differentiation of Bacillus anthracis from other Bacillus cereus group bacteria with the PCR. Int J Syst Bacteriol. 1994 Jan;44(1):99–105. doi: 10.1099/00207713-44-1-99. [DOI] [PubMed] [Google Scholar]
  8. Huys G., Coopman R., Janssen P., Kersters K. High-resolution genotypic analysis of the genus Aeromonas by AFLP fingerprinting. Int J Syst Bacteriol. 1996 Apr;46(2):572–580. doi: 10.1099/00207713-46-2-572. [DOI] [PubMed] [Google Scholar]
  9. Jeffreys A. J., Wilson V., Thein S. L. Individual-specific 'fingerprints' of human DNA. Nature. 1985 Jul 4;316(6023):76–79. doi: 10.1038/316076a0. [DOI] [PubMed] [Google Scholar]
  10. Kaspar R. L., Robertson D. L. Purification and physical analysis of Bacillus anthracis plasmids pXO1 and pXO2. Biochem Biophys Res Commun. 1987 Dec 16;149(2):362–368. doi: 10.1016/0006-291x(87)90375-5. [DOI] [PubMed] [Google Scholar]
  11. Knisely R. F. Selective medium for Bacillus anthracis. J Bacteriol. 1966 Sep;92(3):784–786. doi: 10.21236/ad0479102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MCDONALD W. C., FELKNER I. C., TURETSKY A., MATNEY T. S. SIMILARITY IN BASE COMPOSITIONS OF DEOXYRIBONUCLEATES FROM SEVERAL STRAINS OF BACILLUS CEREUS AND BACILLUS ANTHRACIS. J Bacteriol. 1963 May;85:1071–1073. doi: 10.1128/jb.85.5.1071-1073.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Taylor T. K., Bashiruddin J. B., Gould A. R. Application of a diagnostic DNA probe for the differentiation of the two types of Mycoplasma mycoides subspecies mycoides. Res Vet Sci. 1992 Sep;53(2):154–159. doi: 10.1016/0034-5288(92)90103-9. [DOI] [PubMed] [Google Scholar]
  14. Turnbull P. C., Hutson R. A., Ward M. J., Jones M. N., Quinn C. P., Finnie N. J., Duggleby C. J., Kramer J. M., Melling J. Bacillus anthracis but not always anthrax. J Appl Bacteriol. 1992 Jan;72(1):21–28. doi: 10.1111/j.1365-2672.1992.tb04876.x. [DOI] [PubMed] [Google Scholar]
  15. Vos P., Hogers R., Bleeker M., Reijans M., van de Lee T., Hornes M., Frijters A., Pot J., Peleman J., Kuiper M. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res. 1995 Nov 11;23(21):4407–4414. doi: 10.1093/nar/23.21.4407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Welkos S. L. Plasmid-associated virulence factors of non-toxigenic (pX01-) Bacillus anthracis. Microb Pathog. 1991 Mar;10(3):183–198. doi: 10.1016/0882-4010(91)90053-d. [DOI] [PubMed] [Google Scholar]

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