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Subject: Biotechnology and Molecular Biology × Author: Saha, Abhigyan × End date: 2009 × Type: Thesis × Thesis Type: M.Sc ×
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    ThesisItemOpen Access
    Identification of gene for ADP-Glucose pyrophosphorylase from developing grains of wheat
    (CCSHAU, 2006) Saha, Abhigyan; Sikka, Virendera K.
    A step towards isolation of gene(s) has been undertaken in the proposed study. In order to achieve the target of AGPase cDNA gene library preparation, first of all total RNA was isolated from developing grains (14 DAA), of wheat varieties C306 and WH711. Modified RNA extraction protocol was employed, which gave fairly high quality RNA. Good quality mRNA was fractionated from total RNA using poly AT tract magnetic beads mRNA isolation system (Promega). Complementary DNA (cDNA) library was generated from the mRNA of C306 and WH711 using Lambda zap cDNA kit (Stratagene). Plaque lifts were hybridized with rice AGPase gene probe to identify clones containing wheat AGPase gene. However, exact clone could not be isolated. AGPase enzyme assay was carried out in wheat varieties C306, WH711, WH147M, SG70, MLU-2, at 14DAA and 28DAA. The enzyme activity showed an association with rate of starch accumulation and grain size and yield.
  • Thumbnail Image
    ThesisItemOpen Access
    Identification of gene for ADP-Glucose pyrophosphorylase from developing grains of wheat
    (CCSHAU, 2006) Saha, Abhigyan; Sikka, Virendera K.
    A step towards isolation of gene(s) has been undertaken in the proposed study. In order to achieve the target of AGPase cDNA gene library preparation, first of all total RNA was isolated from developing grains (14 DAA), of wheat varieties C306 and WH711. Modified RNA extraction protocol was employed, which gave fairly high quality RNA. Good quality mRNA was fractionated from total RNA using poly AT tract magnetic beads mRNA isolation system (Promega). Complementary DNA (cDNA) library was generated from the mRNA of C306 and WH711 using Lambda zap cDNA kit (Stratagene). Plaque lifts were hybridized with rice AGPase gene probe to identify clones containing wheat AGPase gene. However, exact clone could not be isolated. AGPase enzyme assay was carried out in wheat varieties C306, WH711, WH147M, SG70, MLU-2, at 14DAA and 28DAA. The enzyme activity showed an association with rate of starch accumulation and grain size and yield.