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Journal of Bacteriology, June 2000, p. 3490-3497, Vol. 182, No. 12
The Sidney Kimmel Cancer Center, San Diego,
California 92121
Received 29 November 1999/Accepted 31 March 2000
We report an analysis of a sample of the SOS response of
Salmonella enterica serovar Typhimurium using the
differential display of RNA fingerprinting gels of arbitrarily primed
PCR products. The SOS response was induced by the addition of mitomycin
C to an exponentially growing culture of serovar Typhimurium, and the RNA population was sampled during the following 2 h. These
experiments revealed 21 differentially expressed PCR fragments
representing mRNA transcripts. These 21 fragments correspond to 20 distinct genes. All of these transcripts were positively regulated,
with the observed induction starting 10 to 120 min after addition of mitomycin C. Fifteen of the 21 transcripts have no homologue in the
public sequence data banks and are therefore classified as novel. The
remaining six transcripts corresponded to the recE, stpA, sulA, and umuC genes, and to
a gene encoding a hypothetical protein in the Escherichia coli
lysU-cadA intergenic region; the recE gene was
represented twice by nonoverlapping fragments. In order to determine if
the induction of these 20 transcripts constitutes part of a classical
SOS regulon, we assessed the induction of these genes in a
recA mutant. With one exception, the increased expression
of these genes in response to mitomycin C was dependent on the presence
of a functional recA allele. The exception was fivefold
induced in the absence of a functional RecA protein, suggesting another
layer of regulation in response to mitomycin C, in addition to the
RecA-LexA pathway of SOS induction. Our data reveal several genes
belonging to operons known to be directly involved in pathogenesis. In
addition, we have found several phage-like sequences, some of which may
be landmarks of pathogenicity determinants. On the basis of these
observations, we propose that the general use of DNA-damaging agents
coupled with differential gene expression analysis may be a useful and
easy method for identifying pathogenicity determinants in diverse organisms.
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Analysis of the SOS Response in Salmonella
enterica Serovar Typhimurium Using RNA Fingerprinting by
Arbitrarily Primed PCR
*
Corresponding author. Mailing address: The Sidney
Kimmel Cancer Center, 10835 Altman Row, San Diego, CA 92121. Phone:
(858) 450-5990, ext. 280. Fax: (858) 550-3998. E-mail:
mmcclelland{at}skcc.org.
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