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Analysis of nuclear proteins from silk glands ofBombyx mori
Authors:Pushpa Agrawal  K P Gopinathan
Institution:(1) Centre for Genetic Engineering and Department of Microbiology and Cell Biology, Indian Institute of Science, 560 012 Bangalore, India
Abstract:A gentle method for the isolation of nuclei from developing silk glands ofBombyx mori has been standardized. The nuclei, whether isolated or directly visualizedin situ within the silk glands, exhibit complex morphology. The nuclei occupy almost the entire volume of the gigantic silk gland cells. Although the isolated nuclei still retain their ramified morphology, being polyploid they are fragile and often become fragmented. The histone and low-salt-extractable proteins from nuclei isolated from the middle and posterior silk glands on different days of the fourth and fifth instars of larval development have been analysed. The histones did not show any stage- or tissue-specific variations whereas the low-salt-extractable proteins showed some developmental stage specific variation. Using the antibody raised against one such protein, its absence in the early stage of development has been confirmed by Western blotting techniques. This developmental stage specific protein may be functionally linked to some activities responsible for boosting up the production of silk or silk-related proteins during the fifth instar of larval development.
Keywords:Nuclei isolation  ramified nuclei  developmental stage specific proteins  histones  silk gland nuclei  silk gland proteins
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