Cloning and characterization of high molecular weight glutenin gene from Triticum aestivum cultivar Dacke
Keywords:Dough, Dacke, HMW glutenin
Background: High molecular weight (HMW) glutenin protein plays a crucial role in determining dough viscoelastic properties that determines the quality of wheat flour. The aim of the present study was to isolate, clone and analyze (in silico) the HMW glutenin gene of Triticum aestivum cultivar Dacke.
Methods: Primers were designed to amplify a 2445 bp fragment of HMW glutenin gene. Ax type HMW glutenin gene from Triticum aestivum cultivar Dacke was isolated using PCR and it was sequenced by primer walking.
Results: Amplified HMW glutenin gene was designated as HMWGAx. Sequence analysis revealed a complete open reading frame encoding 815 amino acid residues with N- and C terminal non-repetitive domain and a central repetitive domain. The calculated molecular weight of the deduced HMW glutenin protein was ~88 kDa and the number of cysteine residues in the HMWGAx was four, in accordance with other x type HMW glutenin proteins. Phylogenetic analysis revealed 100% homology to the previously studied Ax2* type HMW glutenin gene from cultivar Cheyenne. Predicted secondary structure results showed that it was similar to1Ax1 type of common wheat (Triticum aestivum), having superior flour milling quality.
Conclusions: Sequence analysis suggests that HMWGAx protein significantly and positively correlates with the properties of elasticity and extensibility of gluten.
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