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Journal of Bacteriology, October 1999, p. 6509-6515, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Sequence and Organization of pXO1, the Large Bacillus anthracis Plasmid Harboring the Anthrax Toxin Genes

R. T. Okinaka,1,* K. Cloud,1,dagger O. Hampton,1 A. R. Hoffmaster,2 K. K. Hill,1 P. Keim,3 T. M. Koehler,2 G. Lamke,1,Dagger S. Kumano,1,§ J. Mahillon,4 D. Manter,1,parallel Y. Martinez,1 D. Ricke,1 # R. Svensson,1 and P. J. Jackson1

Life Sciences Division, Los Alamos National Laboratory, Los Alamos, New Mexico 875451; Department of Biology, Northern Arizona University, Flagstaff, Arizona 86011-56403; Department of Microbiology and Molecular Genetics, University of Texas---Houston Medical School, Houston, Texas 770302; and Laboratoire de Génétique Microbienne---UCL, Louvain-la-Neuve, Belgium4

Received 12 April 1999/Accepted 30 July 1999

The Bacillus anthracis Sterne plasmid pXO1 was sequenced by random, "shotgun" cloning. A circular sequence of 181,654 bp was generated. One hundred forty-three open reading frames (ORFs) were predicted using GeneMark and GeneMark.hmm, comprising only 61% (110,817 bp) of the pXO1 DNA sequence. The overall guanine-plus-cytosine content of the plasmid is 32.5%. The most recognizable feature of the plasmid is a "pathogenicity island," defined by a 44.8-kb region that is bordered by inverted IS1627 elements at each end. This region contains the three toxin genes (cya, lef, and pagA), regulatory elements controlling the toxin genes, three germination response genes, and 19 additional ORFs. Nearly 70% of the ORFs on pXO1 do not have significant similarity to sequences available in open databases. Absent from the pXO1 sequence are homologs to genes that are typically required to drive theta replication and to maintain stability of large plasmids in Bacillus spp. Among the ORFs with a high degree of similarity to known sequences are a collection of putative transposases, resolvases, and integrases, suggesting an evolution involving lateral movement of DNA among species. Among the remaining ORFs, there are three sequences that may encode enzymes responsible for the synthesis of a polysaccharide capsule usually associated with serotype-specific virulent streptococci.

* Corresponding author. Mailing address: Life Sciences Division, Los Alamos National Laboratory, M888, Los Alamos, NM 87545. Phone: (505) 667-2743. Fax: (505) 665-3024. E-mail: okinaka{at}telomere.lanl.gov.

dagger Present address: University of Colorado Health Science Center, Denver, CO 80220.

Dagger Present address: Mayo Medical School, Rochester, MN 55905.

§ Present address: Department of Molecular and Cell Genetics, Tottori University, Yonago, Tottori 683, Japan.

parallel Present address: College of Forestry, Oregon State University, Corvallis, OR 97331.

Journal of Bacteriology, October 1999, p. 6509-6515, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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