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Journal of Bacteriology, April 1999, p. 2358-2362, Vol. 181, No. 8
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Genetic Diversity in the Protective Antigen Gene of Bacillus anthracis

Lance B. Price,1 Martin Hugh-Jones,2 Paul J. Jackson,3 and Paul Keim1,*

Department of Biological Science, Northern Arizona University, Flagstaff, Arizona 86011-56401; Department of Epidemiology and Community Health, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana 708032; and Environmental Molecular Biology Group, Los Alamos National Laboratory, Los Alamos, New Mexico 875453

Received 2 December 1998/Accepted 29 January 1999

Bacillus anthracis is a gram-positive spore-forming bacterium that causes the disease anthrax. The anthrax toxin contains three components, including the protective antigen (PA), which binds to eucaryotic cell surface receptors and mediates the transport of toxins into the cell. In this study, the entire 2,294-nucleotide protective antigen gene (pag) was sequenced from 26 of the most diverse B. anthracis strains to identify potential variation in the toxin and to further our understanding of B. anthracis evolution. Five point mutations, three synonymous and two missense, were identified. These differences correspond to six different haploid types, which translate into three different amino acid sequences. The two amino acid changes were shown to be located in an area near a highly antigenic region critical to lethal factor binding. Nested primers were used to amplify and sequence this same region of pag from necropsy samples taken from victims of the 1979 Sverdlovsk incident. This investigation uncovered five different alleles among the strains present in the tissues, including two not seen in the 26-sample survey. One of these two alleles included a novel missense mutation, again located just adjacent to the highly antigenic region. Phylogenetic (cladistic) analysis of the pag corresponded with previous strain grouping based on chromosomal variation, suggesting that plasmid evolution in B. anthracis has occurred with little or no horizontal transfer between the different strains.

* Corresponding author. Mailing address: Department of Biological Sciences, Northern Arizona University, Flagstaff, AZ 86011-5640. Phone: (520) 523-1078. Fax: (520) 523-7500. E-mail: Paul.Keim{at}nau.edu.

Journal of Bacteriology, April 1999, p. 2358-2362, Vol. 181, No. 8
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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