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Poster communications

Molecular cloning of cDNA coding for wheat gliadins (PQ-1-001 Session Nuclear Genes)

Abstract : Gliadins, prolamins which constitute the major storage proteins of the wheat (Triticum aestivum L.) endosperm, belong to a multigenic family (several dozens of genes located on different chromosomes). They are all synthesized synchronously and undergo a signal peptide removal when crossind the RER membrane in the lumen of which they transit prior deposition within protein bodies. Only membrane-bound polysomes prime the synthesis of gliadins and comprise 75 superabundant sequences, representing 60% of the mRNA molecules being translated. We have isolated the mRNAs from these polysomes obtained from seeds(cv Cappelle) harvested when the gliadin synhesis is maximal (about 3 weeks after anthesis). These mRNAs were reversely transcribed in double stranded cDNAs in presence of RNAsin, themselves cloned at the Pst I site of pBR322. Recombinant clones were screened following several criteria: size of the insert; size of the mRNA hybridizing the recombinant plasmid; properties of corresponding in vitro translated proteins, i.e. the prolamin-specific solubility in 60% (w/w) ethanol and the SDS-PAGE electrophoretic pattern. Out of a hundred clones, a dozen were selected and submitted to a restriction site analysis. Two clones corresponding to omega- and gamma-gliadins were subcloned in the M3 vector to determine their sequence by the Sanger dideoxy method. The nucleotide and the deduced aminoacid sequences, and also the predicted secondary structure, are discussed in relation to other seed stotage proteins.
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Poster communications
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https://hal.inrae.fr/hal-03152416
Contributor : Victor Vaillant <>
Submitted on : Thursday, February 25, 2021 - 3:39:27 PM
Last modification on : Tuesday, March 23, 2021 - 10:56:02 PM

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  • HAL Id : hal-03152416, version 1

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Victor Vaillant, Christine Camilleri, Pernollet Jc.. Molecular cloning of cDNA coding for wheat gliadins (PQ-1-001 Session Nuclear Genes). First International Congress of Plant Molecular Biology, Oct 1985, Savanah (Georgia), United States. ⟨hal-03152416⟩

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