The sequence of a pea vicilin gene and its expression in transgenic tobacco plants |
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Authors: | Thomas J. V. Higgins Edward J. Newbigin Donald Spencer Danny J. Llewellyn Stuart Craig |
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Affiliation: | (1) Division of Plant Industry, CSIRO, GPO Box 1600, 2601 Canberra, Australia;(2) Dept. Botany, Australian National University, GPO Box 4, 2601 Canberra, Australia |
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Abstract: | A 5.5 kb Eco RI fragment containing a vicilin gene was selected from a Pisum sativum genomic library, and the protein-coding region and adjacent 5 and 3 regions were sequenced. A DNA construction comprising this 5.5 kb fragment together with a gene for neomycin phosphotransferase II was stably introduced into tobacco using an Agrobacterium tumefaciens binary vector, and the fidelity of expression of the pea vicilin gene in its new host was studied. The seeds of eight transgenic tobacco plants showed a sixteen-fold range in the level of accumulated pea vicilin. The level of accumulation of vicilin protein and mRNA correlated with the number of integrated copies of the vicilin gene. Pea vicilin was confined to the seeds of transgenic tobacco. Using immunogold labelling, vicilin was detected in protein bodies of eight out of ten embryos (axes plus cotyledons) and, at a much lower level, in two out of eleven endosperms. Pea vicilin was synthesized early in tobacco seed development; some molecules were cleaved as is the case in pea seeds, yielding a major parental component of Mr50000 together with a range of smaller polypeptides. |
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Keywords: | vicilin gene transgenic tobacco Agrobacterium tumefaciens |
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