Nottingham Trent University
TITLE: CHARACTERISATION OF TRANSGLUTAMINSE IN THE GERMINATAING WHEAT ALEURONE LAYERS
To determine the activity of Transglutaminase in germinating wheat aleuronic layers, extracted samples were prepared from the germinating wheat aleuronic layers and Transglutaminse (Tgase) specific activity and presence of Tgases was determined for the extracted samples. By different procedures we identified Transglutaminase from the extracted samples.This was achieved by the following procedures 1. Samples were extracted in liquid nitrogen, 2. Extracted samples were assayed by Biotin Caderevine assay and protein assay to check the specific activity Tgase, 3. Blotting the samples on nitrocellulose (NC) membrane and identifying the cross reactivity of the extracted Tgases by using specific HRP conjugated immunoglobulin G (IgG) anti-mouse antibody. 4. Antibody specific to the Tgase was identified by testing the standard and concentrated extracted samples by blotting on NC membrane and cross reactivating with different antibodies. 5. Checking out the activity of PA’s by histochemical localisation of the enzyme in the germinated aleuronic layers of the wheat seeds. The activity of the Tgase was found to be increased in the Ca2+ ions. Increase in the activity of the Tgases in Gibberellic acid was found and it was more than the Ca2+ ions. The molecular weight of the Tgase was predicted by SDS-PAGE. Antibody specific to the Tgases was identified by using dot blot and the presence of Tgases in the aleuronic layers was demonstrated by histochemical staining.
Key words: Wheat, aleuronic layers, Transglutaminase (Tgase), Gibberellic acid (GA)