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  • Title: Nucleotide sequence and endosperm-specific expression of the structural gene for the toxin alpha-hordothionin in barley (Hordeum vulgare L.).
    Author: Rodríguez-Palenzuela P, Pintor-Toro JA, Carbonero P, García-Olmedo F.
    Journal: Gene; 1988 Oct 30; 70(2):271-81. PubMed ID: 2850969.
    Abstract:
    A barley genomic library, obtained by cloning in the vector lambda EMBL-4, was screened with a cDNA probe encoding the alpha-hordothionin toxin. A positive clone, designated lambda TH1, was selected for further characterization. The coding and flanking regions of the alpha-hordothionin gene (Hth-1) were sequenced. Hth-1 has two introns of 420 and 91 nucleotides (nt), respectively. The promoter region has the following main features: one TATA box; three CATC boxes; an enhancer-like sequence, starting at nt position -282 from the first ATG codon, which is homologous to sequences appearing at similar positions in other endosperm genes; two versions of an 18-nt sequence that is more highly repeated in structural domains of several prolamin genes; two extensive regions close to the first ATG codon that are homologous to a sequence located much further upstream in the B-hordein promoter. The transcription start point was determined at nt positions -46 to -47, both by the S1 nuclease-protection and by the primer-extension assays. A maximum of 2-4 copies of the Hth-1 gene per haploid genome was determined by Southern-blot hybridization. Expression of the Hth-1 gene was detected during the cell proliferation stage of endosperm development (maximum at 13-16 days after pollinization) and was not detected in either etiolated or green coleoptiles.
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