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Pea convicilin: structure and primary sequence of the protein and expression of a gene in the seeds of transgenic tobacco

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Abstract

Convicilin, a trimeric globulin of pea (Pisum sativum L.) seeds, is closely related to vicilin and composed of polypeptides of 68.2 kilodaltons. A partial copy DNA (cDNA) clone encoding convicilin was isolated, sequenced, and used to select a convicilin gene from a pea genomic library. A part of the genomic clone was sequenced to obtain the coding sequences missing in the cDNA clone and a further 1 kilobase 5′ to the start of transcription were also obtained. The entire sequence of convicilin was deduced from the combined genomic and cDNA sequences. The complete gene encoding convicilin was transferred to tobacco (Nicotiana tabacum L.) and the characteristics of its expression in the seeds of transgenic plants were studied. An unprocessed polypeptide, which was found only in the seeds of the transgenic plants, was identical in size to pea convicilin, and was recognized by vicilin antibodies. Convicilin, which does not undergo posttranslational cleavage in peas, was partially processed to polypeptides of a relative molecular mass (Mr) of approx. 50000 in transgenic tobacco seeds. There was a twofold variation in the level of convicilin accumulated by the mature seeds of a number of transgenic plants and this was well correlated with the number of gene copies incorporated in the different transformants. In the seeds of tobacco plants that contained a single copy of the transferred gene it was estimated that convicilin comprised up to 2% of the seed protein. Thus, using a combination of gene sequencing and expression in a heterologous host we believe we have characterized the gene corresponding to theCvc locus, whereas the gene described by D. Bown et al. (1988, Biochem J.,251, 717–726) probably encodes a minor convicilin-related protein.

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Abbreviations

bp:

base pairs

cDNA:

copy DNA

DAF:

days after flowering

kb:

kilobase pair

kDa:

kilodalton

Mr :

relative molecular mass

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SSC:

standard saline citrate

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Author information

Author notes

  1. Edward J. Newbigin

    Present address: Department of Biochemistry and Biophysics, University of California, 95616, Davis, CA, USA

  2. Ben O. deLumen

    Present address: Department of Nutritional Sciences, University of California, 94720, Berkeley, CA, USA

  3. Alan Gould

    Present address: United Agriseeds, Inc., PO Box 4011, 61820, Champaign, IL, USA

Authors and Affiliations

  1. CSIRO Division of Plant Industry, GPO Box 1600, 2601, Canberra, A.C.T., Australia

    Edward J. Newbigin, Ben O. deLumen, Peter M. Chandler, Alan Gould & Thomas J. V. Higgins

  2. Department of Botany, Australian National University, GPO Box 4, 2601, Canberra, A.C.T., Australia

    Edward J. Newbigin

  3. CSIRO Division of Biotechnology, 343 Royal Pde, 3052, Parkyille, Victoria, Australia

    Robert J. Blagrove & Alex A. Kortt

  4. John Innes Institute, Colney Lane, NR4 7UH, Norwich, UK

    John F. March

Authors
  1. Edward J. Newbigin
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  8. Thomas J. V. Higgins
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Newbigin, E.J., deLumen, B.O., Chandler, P.M. et al. Pea convicilin: structure and primary sequence of the protein and expression of a gene in the seeds of transgenic tobacco. Planta 180, 461–470 (1990). https://doi.org/10.1007/BF02411442

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  • DOI: https://doi.org/10.1007/BF02411442

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