Messenger RNA from isolated aleurone cells directs the synthesis of an alpha-galactosidase found in the endosperm during germination of guar (Cyamopsis tetragonaloba) seed |
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Authors: | Stephen G Hughes Nico Overbeeke Susan Robinson Karen Pollock Frans L M Smeets |
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Institution: | (1) Unilever Research Laboratory Colworth House, Sharnbrook, United Kingdom;(2) Unilever Research Laboratorium Vlaardingen, Vlaardingen, Netherlands;(3) Unilever Research Laboratorium, P.O. Box 114, 3130 AC Vlaardingen, Netherlands |
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Abstract: | In cereals and some legumes the aleurone layer is a site of synthesis of enzymes which mobilize endosperm reserves. It has been established whether or not the aleurone cells of the seed endosperm of Cyamopsis tetragonaloba are a site of synthesis of -galactosidase. The isolation and cultivation of aleurone cells demonstrated that they contain mRNA which directs the synthesis and secretion of -galactosidase into the endosperm where along with a -mannanase it is responsible for the degradation of the galactomannan storage polymer. A method was developed to purify the mRNA from the aleurone cells of germinating seeds. This mRNA was analysed by: i) Northern blot hybridization using oligo-nucleotide mixed probes derived from the protein's NH2-terminal amino acid sequence and ii) in vitro translation in a wheat germ system and detection of the -galactosidase protein using antibodies. The molecular mass of the protein synthesized in vitro is slightly larger (44 kDa) than that of the mature -galactosidase (40.5 kDa) which is as expected for the precursor of a secreted protein. |
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Keywords: | -galactosidase" target="_blank">gif" alt="agr" align="BASELINE" BORDER="0">-galactosidase aleurone cells Cyamopsis tetragonaloba guar |
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