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Applied and Environmental Microbiology, May 2003, p. 2755-2764, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2755-2764.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Genome Differences That Distinguish Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis

Lyndsay Radnedge,1 Peter G. Agron,1 Karen K. Hill,2 Paul J. Jackson,2 Lawrence O. Ticknor,2 Paul Keim,3 and Gary L. Andersen4*

Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California 94551,1 Bioscience Division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545,2 Department of Biological Sciences, Northern Arizona University, Flagstaff, Arizona 86011;,3 Center for Environmental Biotechnology, Lawrence Berkeley National Laboratory, Berkeley, California 947204

Received 15 October 2002/ Accepted 13 February 2003

The three species of the group 1 bacilli, Bacillus anthracis, B. cereus, and B. thuringiensis, are genetically very closely related. All inhabit soil habitats but exhibit different phenotypes. B. anthracis is the causative agent of anthrax and is phylogenetically monomorphic, while B. cereus and B. thuringiensis are genetically more diverse. An amplified fragment length polymorphism analysis described here demonstrates genetic diversity among a collection of non-anthrax-causing Bacillus species, some of which show significant similarity to B. anthracis. Suppression subtractive hybridization was then used to characterize the genomic differences that distinguish three of the non-anthrax-causing bacilli from B. anthracis Ames. Ninety-three DNA sequences that were present in B. anthracis but absent from the non-anthrax-causing Bacillus genomes were isolated. Furthermore, 28 of these sequences were not found in a collection of 10 non-anthrax-causing Bacillus species but were present in all members of a representative collection of B. anthracis strains. These sequences map to distinct loci on the B. anthracis genome and can be assayed simultaneously in multiplex PCR assays for rapid and highly specific DNA-based detection of B. anthracis.

* Corresponding author. Mailing address: Center for Environmental Biotechnology, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd., Mail Stop 70A-3317, Berkeley, CA 94720. Phone: (510) 495-2795. Fax: (510) 486-7152. E-mail: GLAndersen{at}lbl.gov.

Applied and Environmental Microbiology, May 2003, p. 2755-2764, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2755-2764.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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