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Applied and Environmental Microbiology, November 2009, p. 7163-7172, Vol. 75, No. 22
0099-2240/09/$08.00+0     doi:10.1128/AEM.01069-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Identification and Classification of bcl Genes and Proteins of Bacillus cereus Group Organisms and Their Application in Bacillus anthracis Detection and Fingerprinting{triangledown} ,{dagger}

Tomasz A. Leski,1 Clayton C. Caswell,2,{ddagger} Marcin Pawlowski,5 David J. Klinke,2,3 Janusz M. Bujnicki,5,6 Sean J. Hart,7 and Slawomir Lukomski2,4*

Nova Research, Inc., Alexandria, Virginia 22308,1 Department of Microbiology, Immunology, and Cell Biology,2 Department of Chemical Engineering,3 Mary Babb Randolph Cancer Center, West Virginia University, Morgantown, West Virginia 26506,4 Laboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell Biology, 02-109 Warsaw,5 Bioinformatics Laboratory, Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University, 61-614 Poznan, Poland,6 Bio/Analytical Chemistry, Chemistry Division, Code 6112, Naval Research Laboratory, Washington, DC 203757

Received 9 May 2009/ Accepted 10 September 2009

The Bacillus cereus group includes three closely related species, B. anthracis, B. cereus, and B. thuringiensis, which form a highly homogeneous subdivision of the genus Bacillus. One of these species, B. anthracis, has been identified as one of the most probable bacterial biowarfare agents. Here, we evaluate the sequence and length polymorphisms of the Bacillus collagen-like protein bcl genes as a basis for B. anthracis detection and fingerprinting. Five genes, designated bclA to bclE, are present in B. anthracis strains. Examination of bclABCDE sequences identified polymorphisms in bclB alleles of the B. cereus group organisms. These sequence polymorphisms allowed specific detection of B. anthracis strains by PCR using both genomic DNA and purified Bacillus spores in reactions. By exploiting the length variation of the bcl alleles it was demonstrated that the combined bclABCDE PCR products generate markedly different fingerprints for the B. anthracis Ames and Sterne strains. Moreover, we predict that bclABCDE length polymorphism creates unique signatures for B. anthracis strains, which facilitates identification of strains with specificity and confidence. Thus, we present a new diagnostic concept for B. anthracis detection and fingerprinting, which can be used alone or in combination with previously established typing platforms.

* Corresponding author. Mailing address: Department of Microbiology, Immunology, and Cell Biology, West Virginia University HSC, 2095 Health Sciences North, Box 9177, Morgantown, WV 26506. Phone: (304) 293-6405. Fax: (304) 293-7823. E-mail: slukomski{at}hsc.wvu.edu

{triangledown} Published ahead of print on 18 September 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

{ddagger} Present address: Department of Microbiology and Immunology, Brody School of Medicine, East Carolina University, Greenville, NC.

Applied and Environmental Microbiology, November 2009, p. 7163-7172, Vol. 75, No. 22
0099-2240/09/$08.00+0     doi:10.1128/AEM.01069-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Tan, L., Turnbough, C. L. Jr. (2010). Sequence Motifs and Proteolytic Cleavage of the Collagen-Like Glycoprotein BclA Required for Its Attachment to the Exosporium of Bacillus anthracis. J. Bacteriol. 192: 1259-1268 [Abstract] [Full Text]  


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