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Journal of Bacteriology, October 2000, p. 5692-5699, Vol. 182, No. 20
Department of Plant Biology, Carnegie Institution of
Washington, Stanford, California 943051;
Station Biologique, CNRS, INSU et Université Pierre et Marie
Curie, Roscoff Cedex, France2; and
Calydon, Sunnyvale, California 940893
Received 27 April 2000/Accepted 18 July 2000
Global identification of differentially regulated genes in
prokaryotes is constrained because the mRNA does not have a 3' polyadenylation extension; this precludes specific separation of mRNA
from rRNA and tRNA and synthesis of cDNAs from the entire mRNA
population. Knowledge of the entire genome sequence of
Synechocystis sp. strain PCC 6803 has enabled us to develop
a differential display procedure that takes advantage of a short
palindromic sequence that is dispersed throughout the
Synechocystis sp. strain PCC 6803 genome. This sequence,
designated the HIP (highly iterated palindrome) element, occurs in
approximately half of the Synechocystis sp. strain PCC
6803 genes but is absent in rRNA and tRNA genes. To determine the
feasibility of exploiting the HIP element, alone or in combination with
specific primer subsets, for analyzing differential gene expression, we
used HIP-based primers to identify light intensity-regulated genes.
Several gene fragments, including those encoding ribosomal proteins and
phycobiliprotein subunits, were differentially amplified from RNA
templates derived from cells grown in low light or exposed to high
light for 3 h. One novel finding was that expression of certain
genes of the pho regulon, which are under the control of
environmental phosphate levels, were markedly elevated in high light.
High-light activation of pho regulon genes correlated with
elevated growth rates that occur when the cells are transferred from
low to high light. These results suggest that in high light, the rate
of growth of Synechocystis sp. strain PCC 6803 exceeds its
capacity to assimilate phosphate, which, in turn, may trigger a
phosphate starvation response and activation of the pho regulon.
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Isolation of Regulated Genes of the Cyanobacterium
Synechocystis sp. Strain PCC 6803 by Differential
Display
*
Corresponding author. Mailing address: Carnegie
Institution of Washington, Department of Plant Biology, 260 Panama St.,
Stanford, CA 94305. Phone: (650) 325-1521, ext. 282. Fax: (650)
325-6857. E-mail: devaki{at}andrew2.stanford.edu.
Carnegie Institution of Washington publication no. 1436.
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