In:
Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 98, No. 5 ( 2001-02-27), p. 2555-2560
Kurzfassung:
Gene expression profiling provides powerful analyses of
transcriptional responses to cellular perturbation. In contrast to DNA array-based methods, reporter gene technology has been underused for
this application. Here we describe a genomewide, genome-registered collection of Escherichia coli bioluminescent reporter
gene fusions. DNA sequences from plasmid-borne, random fusions of E. coli chromosomal DNA to a Photorhabdus
luminescens luxCDABE reporter allowed precise mapping of each
fusion. The utility of this collection covering about 30% of the transcriptional units was tested by analyzing individual fusions
representative of heat shock, SOS, OxyR, SoxRS, and cya/crp stress-responsive regulons. Each fusion strain
responded as anticipated to environmental conditions known to activate the corresponding regulatory circuit. Thus, the collection mirrors E. coli 's transcriptional wiring diagram. This
genomewide collection of gene fusions provides an independent test of results from other gene expression analyses. Accordingly, a DNA
microarray-based analysis of mitomycin C-treated E. coli indicated elevated expression of expected and unanticipated genes.
Selected luxCDABE fusions corresponding to these
up-regulated genes were used to confirm or contradict the DNA microarray results. The power of partnering gene fusion and DNA
microarray technology to discover promoters and define operons was demonstrated when data from both suggested that a cluster of 20 genes
encoding production of type I extracellular polysaccharide in E.
coli form a single operon.
Materialart:
Online-Ressource
ISSN:
0027-8424
,
1091-6490
DOI:
10.1073/pnas.041620498
Sprache:
Englisch
Verlag:
Proceedings of the National Academy of Sciences
Publikationsdatum:
2001
ZDB Id:
209104-5
ZDB Id:
1461794-8
SSG:
11
SSG:
12
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