Abstract
A subtractive hybridization procedure was used to clone genes of the cyanobacterium Anabaena torulosa expressed in response to salt stress. The method uses total RNA from salt-treated cells, labeled in vitro, as the probe. Hybridization to restriction digests of total DNA was used for interspecies comparison; the procedure was also successful for isolation of cosmids by colony hybridization, semiquantitative dot blots, and cosmid characterization by Southern blotting. Analysis of eleven independent cosmids containing genes whose transcription is abundantly induced by salt suggests that a substantial portion of the A. torulosa genome, probably in excess of 100 kilobases, responds to salt.
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References
Apte SK, Bhagwat AA: Salinity-stress-induced proteins in two nitrogen-fixing Anabaena strains differentially tolerant to salt. J Bact 171: 909–915 (1989).
Apte SK, Reddy BR, Thomas ST: Relationship between sodium influx and salt tolerance of nitrogen-fixing cyanobacteria. Appl Env Microbiol 53: 1934–1939 (1987).
Apte SK, Thomas J: Membrane electrogenesis and sodium transport in filamentous nitrogen-fixing cyanobacteria. Eur J Biochem 154: 395–401 (1986).
Bhagwat AA, Apte SK: Comparative analysis of proteins induced by heat shock, salinity, and osmotic stress in the nitrogen-fixing cyanobacterium Anabaena sp. strain L-31. J Bact 171: 5187–5189 (1989).
Blumwald E, Mehlhorn RJ, Packer L: Studies of osmoregulation in salt adaptation of cyanobacteria with ESR spin probe techniques. Proc Natl Acad Sci USA 80: 2599–2602 (1983).
Brock TD: Evolutionary and ecological aspects of the cyanophytes. In: Carr NG, Whitton BA (eds) The Biology of Blue-Green Algae, pp. 487–500. Blackwell, London (1973).
Cairney J, Booth IR, Higgins F: Salmonella typhimurium proP gene encodes a transport system for the osmoprotectant betaine. J Bact 164: 1218–1223 (1985).
Cairney J, Booth IR, Higgins F: Osmoregulation of gene expression in Salmonella typhimurium:ProU encodes an osmotically induced betaine transport system. J Bact 164: 1224–1232 (1985).
Castenholz RW: Ecology of blue-green algae in hot springs. In: Carr NG, Whitton BA (eds) The Biology of Blue-Green Algae, pp. 379–414. Blackwell, London (1973).
Castenholz RW: Culturing methods for cyanobacteria. Meth Enzymol 167: 68–93 (1988).
Csonka LN: Physiological and genetic responses of bacteria to osmotic stress. Microbiol Rev 53: 121–147 (1989).
Fogg GE: Physiology and ecology of marine blue-green algae. In: Carr NG, Whitton BA (eds) The Biology of Blue-Green Algae, pp. 368–378. Blackwell, London (1973).
Giaever HM, Styrvold OB, Kaasen I, Strom AR: Biochemical and genetic characterization of osmoregulatory trehalose synthesis in E. coli. J Bact 170: 2841–2849 (1988).
Gutierrez C, Barondess J, Manoil C, Beckwith J: The use of transposon TnphoA to detect genes for cell envelope proteins subject to a common regulatory stimulus. J Mol Biol 195: 289–297 (1987).
Jakowec MW, Smith LT, Dandekar AM: Recombinant plasmid conferring proline overproduction and osmotolerance. Appl Env Microbiol 50: 441–446 (1985).
Mackay MA, Norton RS, Borowitzka LJ: Organic osmoregulatory solutes in cyanobacteria. Mar Biol 73: 301–307 (1983).
Mahan MJ, Gsonka LN: Genetic analysis of the proBA genes of Salmonella typhimurium: physical and genetic analyses of the cloned proB +A+genes of E. coli and of a mutant allele that confers proline overproduction and osmotolerance. J Bact 156: 1249–1262 (1983).
Maniatis T, Fritsch FF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Ramirez RM, Prince WS, Bremer E, Villarejo M: In vitro reconstitution of osmoregulated expression of proU of E. coli. Proc Natl Acad Sci USA 86: 1153–1157 (1989).
Reed RH, Chudek JA, Foster R, Stewart WDP: Osmotic adjustments in cyanobacteria from hypersaline environments. Arch Microbiol 138: 333–337 (1984).
Reed RH, Richardson DL, Warr SRC, Stewart WDP: Carbohydrate accumulation and osmotic stress in cyanobacteria. J Gen Mirobiol 130: 1–4 (1984).
Roger PA, Watanabe I: Technologies for utilizing biological nitrogen fixation in wetland rice: potentialities, current usage, and limiting factors. Fert Res 9: 39–77 (1986).
Schlessinger MJ, Ashburner M, Tissieres A: Heat Shock from Bacteria to Man. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Strom AR, Falkenberg P, Landfald B: Genetics of osmoregulation in E. coli: uptake and biosynthesis of organic osmolytes. FEMS Microbiol Rev 39: 79–86 (1986).
Styrvold OB, Falkenberg P, Landfald B, Eshoo MW, Bjornsen T, Strom AR: Selection, mapping, and characterization of osmoregulatory mutants of E. coli blocked in the choline-glycine betaine pathway. J Bact 165: 856–863 (1986).
Walderhaug MO, Dosch DC, Epstein E: Potassium transport in bacteria. In: Rosen BP, Silver S (eds) Ion Transport in Prokaryotes, pp. 85–130 (1987).
Walderhaug MO, Litwack ED, Epstein W: Wide distribution of homologs of E. coli Kdp K+-ATPase among gram-negative bacteria. J Bact 171: 1191–1195 (1989).
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Apte, S.K., Haselkorn, R. Cloning of salinity stress-induced genes from the salt-tolerant nitrogen-fixing cyanobacterium Anabaena torulosa . Plant Mol Biol 15, 723–733 (1990). https://doi.org/10.1007/BF00016122
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DOI: https://doi.org/10.1007/BF00016122