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Applied and Environmental Microbiology, July 2000, p. 2842-2852, Vol. 66, No. 7
Swiss Federal Institute for Environmental
Science and Technology1 and Swiss
Federal Institute of Technology,2 CH-8600
Dübendorf, Switzerland, and Institute of Genetics and
Cytology, National Academy of Sciences of Belarus, 220072 Minsk,
Belarus3
Received 24 January 2000/Accepted 11 April 2000
Twenty different Pseudomonas strains utilizing
m-toluate were isolated from oil-contaminated soil samples
near Minsk, Belarus. Seventeen of these isolates carried plasmids
ranging in size from 78 to about 200 kb (assigned pSVS plasmids) and
encoding the meta cleavage pathway for toluene metabolism.
Most plasmids were conjugative but of unknown incompatibility groups,
except for one, which belonged to the IncP9 group. The organization of
the genes for toluene catabolism was determined by restriction analysis
and hybridization with xyl gene probes of pWW0. The
majority of the plasmids carried xyl-type genes highly
homologous to those of pWW53 and organized in a similar manner (M. T. Gallegos, P. A. Williams, and J. L. Ramos, J. Bacteriol.
179:5024-5029, 1997), with two distinguishable meta
pathway operons, one upper pathway operon, and three
xylS-homologous regions. All of these plasmids also
possessed large areas of homologous DNA outside the catabolic genes,
suggesting a common ancestry. Two other pSVS plasmids carried only one
meta pathway operon, one upper pathway operon, and one copy
each of xylS and xylR. The backbones of these
two plasmids differed greatly from those of the others. Whereas these
parts of the plasmids, carrying the xyl genes, were mostly
conserved between plasmids of each group, the noncatabolic parts had
undergone intensive DNA rearrangements. DNA sequencing of specific
regions near and within the xylTE and xylA
genes of the pSVS plasmids confirmed the strong homologies to the
xyl genes of pWW53 and pWW0. However, several
recombinations were discovered within the upper pathway operons of the
pSVS plasmids and pWW0. The main genetic mechanisms which are thought
to have resulted in the present-day configuration of the
xyl operons are discussed in light of the diversity
analysis carried out on the pSVS plasmids.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Molecular Diversity of Plasmids Bearing Genes That
Encode Toluene and Xylene Metabolism in Pseudomonas Strains
Isolated from Different Contaminated Sites in Belarus
*
Corresponding author. Mailing address: EAWAG,
Department of Microbiology, Überlandstrasse 133, CH-8600
Dübendorf, Switzerland. Phone: 41-1-823 53 24. Fax: 41-1-823 55 47. E-mail: sentchilo{at}eawag.ch.
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