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Journal of Bacteriology, October 2000, p. 5530-5538, Vol. 182, No. 19
Howard Hughes Medical Institute and Division
of Geographic Medicine and Infectious Diseases, Tufts-New England
Medical Center, Tufts University School of Medicine, Boston,
Massachusetts 02111
Received 10 March 2000/Accepted 10 July 2000
The genes encoding cholera toxin (ctxA and
ctxB) are encoded in the genome of CTX
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Molecular Analyses of a Putative CTX
Precursor and Evidence
for Independent Acquisition of Distinct CTX
s by
Toxigenic Vibrio cholerae

, a filamentous
phage that infects Vibrio cholerae. To study the
evolutionary history of CTX
, we examined genome diversity in CTX
s
derived from a variety of epidemic and nonepidemic Vibrio
sp. natural isolates. Among these were three V. cholerae
strains that contained CTX prophage sequences but not the
ctxA and ctxB genes. These prophages each gave
rise to a plasmid form whose genomic organization was very similar to that of the CTX
replicative form, with the exception of missing ctxAB. Sequence analysis of these three plasmids revealed
that they lacked the upstream control region normally found 5' of
ctxA, as well as the ctxAB promoter region and
coding sequences. These findings are consistent with the
hypothesis that a CTX
precursor that lacked ctxAB
simultaneously acquired the toxin genes and their regulatory sequences.
To assess the evolutionary relationships among additional CTX
s, two
CTX
-encoded genes, orfU and zot, were
sequenced from 13 V. cholerae and 4 V. mimicus
isolates. Comparative nucleotide sequence analyses revealed that the
CTX
s derived from classical and El Tor V. cholerae
isolates comprise two distinct lineages within otherwise nearly
identical chromosomal backgrounds (based on mdh sequences).
These findings suggest that nontoxigenic precursors of the two
V. cholerae O1 biotypes independently acquired
distinct CTX
s.
*
Corresponding author. Mailing address: Division of
Geographic Medicine and Infectious Diseases, New England Medical Center #041, 750 Washington St., Boston, MA 02111. Phone: (617) 636-7618. Fax:
(617) 636-5292. E-mail: mwaldor{at}lifespan.org.
Present address: Department of Microbiology, University College
Cork, Cork, Ireland.
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