Cloning,Sequence Analysis and Putative Promoter Region of a Glyceraldehyde-3-Phosphate Dehydrogenase

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The complete sequence(from ATG to TAA) of a glyceraldehyde-3-phosphate dehydrogenase(gpd) gene isolated from Volvariella volvacea consisted of 1489 bp and contained nine introns of similar size.The full-length cDNA of gpd contained 1157 nucleotides with a predicted ORF of 1011 bp encoding for 337 amino acids and a 3’-untranslated region of 95 bp.Putative promoter elements have been identified in a 1220 bp 5’-flanking region,analysis of which revealed two TATA boxes located 80 and 559 bp upstream of the start codon,and three CAAT boxes 333,340 and 406 bp upstream of the initiating ATG.Southern blot analysis suggested that the V.volvacea genome contained only one copy of the gpd gene.Northern blot analysis revealed that gpd gene expression was not induced by cold stress,which may explain in part why this mushroom is more perishable than other mushrooms in which gpd expression increases during exposure to low temperatures. The complete sequence (from ATG to TAA) of a glyceraldehyde-3-phosphate dehydrogenase (gpd) gene isolated from Volvariella volvacea consisted of 1489 bp and contained nine introns of similar size. The full-length cDNA of gpd contained 1157 nucleotides with a predicted ORF of 1011 bp encoding for 337 amino acids and a 3’-untranslated region of 95 bp. Preative promoter elements have been identified in a 1220 bp 5’-flanking region, analysis of which revealed two TATA boxes located 80 and 559 bp upstream of the start codon, and three CAAT boxes 333, 340 and 406 bp upstream of the initiating ATG. Southern blot analysis suggested that the V. volvacea genome contained only one copy of the gpd gene. Northern blot analysis revealed that gpd gene expression was not induced by cold stress, which may explain in part why this mushroom is more perishable than other mushrooms in which gpd expression increases during exposure to low temperatures.
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