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Journal of Bacteriology, October 2000, p. 5692-5699, Vol. 182, No. 20
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†

Devaki Bhaya,1,* Daniel Vaulot,2 Pinky Amin,3 Akiko Watanabe Takahashi,1 and Arthur R. Grossman1

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.

* 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.

dagger Carnegie Institution of Washington publication no. 1436.

Journal of Bacteriology, October 2000, p. 5692-5699, Vol. 182, No. 20
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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