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Research ArticleMolecular Biology and Gene Regulation
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Cloning, Characterization, and Expression of a cDNA Encoding a 50-Kilodalton Protein Specifically Induced by Cold Acclimation in Wheat

Mario Houde, Jean Danyluk, Jean-François Laliberté, Eric Rassart, Rajinder S. Dhindsa, Fathey Sarhan
Mario Houde
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Jean Danyluk
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Jean-François Laliberté
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Eric Rassart
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Rajinder S. Dhindsa
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Fathey Sarhan
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Published August 1992. DOI: https://doi.org/10.1104/pp.99.4.1381

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Abstract

We have isolated, sequenced, and expressed a cold-specific cDNA clone, Wcs120, that specifically hybridizes to a major mRNA species of approximately 1650 nucleotides from cold-acclimated wheat (Triticum aestivum L.). The accumulation of this mRNA was induced in less than 24 hours of cold treatment, and remained at a high steady-state level during the entire period of cold acclimation in the two freezing-tolerant genotypes of wheat tested. The expression of Wcs120 was transient in a less-tolerant genotype even though the genomic organization of the Wcs120 and the relative copy number were the same in the three genotypes. The mRNA level decreased rapidly during deacclimation and was not induced by heat shock, drought, or abscisic acid. The Wcs120 cDNA contains a long open reading frame encoding a protein of 390 amino acids. The encoded protein is boiling stable, highly hydrophilic, and has a compositional bias for glycine (26.7%), threonine (16.7%), and histidine (10.8%), although cysteine, phenylalanine, and tryptophan were absent. The WCS120 protein contains two repeated domains. Domain A has the consensus amino acid sequence GEKKGVMENIKEKLPGGHGDHQQ, which is repeated 6 times, whereas domain B has the sequence TGGTYGQQGHTGTT, which is repeated 11 times. The two domains were also found in barley dehydrins and rice abscisic acid-induced protein families. The expression of this cDNA in Escherichia coli, using the T7 RNA polymerase promoter, produced a protein of 50 kilodaltons with an isoelectric point of 7.3, and this product comigrated with a major protein synthesized in vivo and in vitro during cold acclimation.

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Cloning, Characterization, and Expression of a cDNA Encoding a 50-Kilodalton Protein Specifically Induced by Cold Acclimation in Wheat
Mario Houde, Jean Danyluk, Jean-François Laliberté, Eric Rassart, Rajinder S. Dhindsa, Fathey Sarhan
Plant Physiology Aug 1992, 99 (4) 1381-1387; DOI: 10.1104/pp.99.4.1381

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Cloning, Characterization, and Expression of a cDNA Encoding a 50-Kilodalton Protein Specifically Induced by Cold Acclimation in Wheat
Mario Houde, Jean Danyluk, Jean-François Laliberté, Eric Rassart, Rajinder S. Dhindsa, Fathey Sarhan
Plant Physiology Aug 1992, 99 (4) 1381-1387; DOI: 10.1104/pp.99.4.1381
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Plant Physiology
Vol. 99, Issue 4
August 1992
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More in this TOC Section

  • Cloning of a Temperature-Regulated Gene Encoding a Chloroplast [omega]-3 Desaturase from Arabidopsis thaliana
  • Structural Features of the Maize sus1 Gene and Protein
  • Genes Galore: A Summary of Methods for Accessing Results from Large-Scale Partial Sequencing of Anonymous Arabidopsis cDNA Clones
Show more Molecular Biology and Gene Regulation

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