Skip to main content
SpringerLink
Log in

Cloning and characterization of complementary DNA for convicilin, a major seed storage protein in Pisum sativum L.

  • Published:
Planta Aims and scope Submit manuscript

Abstract

A complementary DNA (cDNA) clone for convicilin, a major storage protein, has been isolated from a cDNA library prepared in the plasmid vector pAT 153, using poly(A)+RNA from developing seeds of Pisum sativum L. The clone was identified by hybrid-selection with poly(A)+ RNA, translation of selected RNAs and immunoprecipitation of the translation products by antibody raised against purified convicilin subunits. The size of the mRNA coding for convicilin polypeptides has been established using this convicilin cDNA clone and has been found to be appreciably longer than the mRNAs coding for the polypeptides of vicilin, a related but separate storage protein. The insert from this convicilin cDNA clone has been compared with the insert from a vicilin cDNA clone by restriction enzyme analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

cDNA:

complementary DNA

IgG:

immunoglobulin G

PAGE:

polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate

Poly(A)+ RNA:

polyadenylated RNA

References

  • Bedbrook, J.R., Smith, S.M., Ellis, R.J. (1980) Molecular cloning and sequencing of cDNA encoding the precursor to the small subunit of chloroplast ribulose-1,5-bisphosphate carboxylase. Nature (London) 287, 692–697

    Google Scholar 

  • Casey, R. (1979a) Immunoaffinity chromatography as a means of purifying legumin from Pisum (pea) seeds. Biochem. J. 177, 509–520

    Google Scholar 

  • Casey, R. (1979b) Genetic variability in the structure of the α-subunits of legumin from Pisum — a two-dimensional gel electrophoresis study. Heredity 43, 265–272

    Google Scholar 

  • Casey, R., Sanger, E. (1980) Purification and some properties of a 7S seed storage protein from Pisum (pea). Biochem. Soc. Trans. 8, 658

    Google Scholar 

  • Chandler, P.M. (1982) The use of single-stranded phage DNAs in hybrid arrest and release translation. Anal. Biochem. 127, 9–16

    Google Scholar 

  • Chrispeels, M.J., Higgins, T.J.V., Spencer, D. (1982) Assembly of storage protein oligomers in the endoplasmic reticulum and processing of the polypeptides in the protein bodies of developing pea cotyledons. J. Cell Biol. 93, 306–313

    Google Scholar 

  • Christophe, D., Brocas, H., Vassart, G. (1982) Improved synthesis of DBM paper. Anal. Biochem. 120, 259–261

    Google Scholar 

  • Covey, S.N., Lomonossoff, G.P., Hull, R. (1981) Characterisation of cauliflower mosaic virus DNA sequences which encode major polyadenylated transcripts. Nucl. Acids Res. 9, 6735–6747

    Google Scholar 

  • Croy, R.R.D., Derbyshire, E., Krishna, T.G., Boulter, D. (1979) Legumin of Pisum sativum and Vicia faba. New Phytol. 83, 29–35

    Google Scholar 

  • Croy, R.R.D., Gatchouse, J.A., Tyler, M., Boulter, D. (1980) The purification and characterization of a third storage protein (convicilin) from the seeds of pea (Pisum sativum L.). Biochem. J. 191, 509–516

    Google Scholar 

  • Danielsson, C.E. (1949) Seed globulins of the Graminae and Leguminosae. Biochem. J. 44, 387–400

    Google Scholar 

  • Davis, R.W., Thomas, M., Cameron, J., St. John, T.P., Scherer, S., Padgett, R.A. (1980) Rapid DNA isolations for enzymatic and hybridization analysis. Methods Enzymol. 65, 404–411

    Google Scholar 

  • Denhardt, D.T. (1966) A membrane-filter technique for the detection of complementary DNA. Biochem. Biophys. Res. Commun. 23, 641–646

    Google Scholar 

  • Domoney, C., Davies, D.R., Casey, R. (1980) The initiation of legumin synthesis in immature embryos of Pisum sativum L. grown in vivo and in vitro. Planta 149, 454–460

    Google Scholar 

  • Domoney, C. (1981) Legumin synthesis and accumulation in Pisum sativum L. Ph. D. thesis, University of East Anglia, Norwich, UK

    Google Scholar 

  • Gatehouse, J.A., Croy, R.R.D., Morton, H., Tyler, M., Boulter, D. (1981) Characterisation and subunit structures of the vicilin storage protein of pea (Pisum sativum L.). Eur. J. Biochem. 118, 627–633

    Google Scholar 

  • Gatehouse, J.A., Evans, I.M., Brown, D., Croy, R.R.D., Boulter, D. (1982) Control of storage-protein synthesis during seed development in pea (Pisum sativum L.). Biochem. J. 208, 119–127

    Google Scholar 

  • Goldsbrough, P.B., Ellis, T.H.N., Lomonossoff, G.P. (1982) Sequence variation and methylation of the flax 5S RNA genes. Nucl. Acids Res. 10, 4501–4514

    Google Scholar 

  • Grunstein, M., Hogness, D.S. (1975) Colony hybridization: A method for the isolation of cloned DNAs that contain a specific gene. Proc. Natl. Acad. Sci. USA 72, 3961–3965

    Google Scholar 

  • Higgins, T.J.V., Spencer, D. (1981) Precursor forms of pea vicilin subunits. Plant Physiol. 67, 205–211

    Google Scholar 

  • Jen, G., Thach, R.E. (1982) Inhibition of host translation in encephalomyocarditis virus-infected L cells: a novel mechanism. J. Virol. 43, 250–261

    Google Scholar 

  • Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227, 680–685

    Google Scholar 

  • Larkins, B.A., Davies, E. (1975) Polyribosomes from peas. V. An attempt to characterize the total free and membrane-bound polysomal population. Plant Physiol. 55, 749–756

    Google Scholar 

  • Martin, C., Northeote, D.H. (1982) The action of exogenous gibberellic acid on isocitrate lyase-mRNA in germinating castor bean seeds. Planta 154, 174–183

    Google Scholar 

  • Matta, N.K., Gatehouse, J.A. (1982) Inheritance and mapping of storage protein genes in Pisum sativum L. Heredity 48, 383–392

    Google Scholar 

  • McMaster, G.K., Carmichael, G.G. (1977) Analysis of single and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc. Natl. Acad. Sci. USA 74, 4835–4838

    Google Scholar 

  • Millerd, A., Thomson, J.A., Schroeder, H.E. (1978) Cotyledonary storage proteins in Pisum sativum. III. Patterns of accumulation during development. Aust. J. Plant Physiol. 5, 519–534

    Google Scholar 

  • Osborne, T.B., Campbell, G.F. (1898) Proteids of the pea. J. Am. Chem. Soc. 20, 348–362, 362–375, 393–405

    Google Scholar 

  • Pelham, H.R.B., Jackson, R.J. (1976) An efficient mRNA-dependent translation system from reticulocyte lysates. Eur. J. Biochem. 67, 247–256

    Google Scholar 

  • Roychoudhury, R., Jay, E., Wu, R. (1976) Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase. Nucl. Acids Res. 3, 101–116

    Google Scholar 

  • Spencer, D., Higgins, T.J.V., Button, S.C., Davey, R.A. (1980) Pulse-labeling studies on protein synthesis in developing pea seeds and evidence of a precursor form of legumin small subunit. Plant Physiol. 66, 510–515

    Google Scholar 

  • Thomas, P.S. (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. USA 77, 5201–5205

    Google Scholar 

  • Thomson, J.A., Schroeder, H.E., Dudman, W.F. (1978) Cotyledonary storage proteins in Pisum sativum. I. Molecular heterogeneity. Aust. J. Plant Physiol. 5, 263–279

    Google Scholar 

  • Weber, K., Osborn, M. (1969) The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J. Biol. Chem. 244, 4406–4412

    Google Scholar 

  • Weber, K., Osborn, M. (1975) Proteins and sodium dodecyl sulfate: molecular weight determination on polyacrylamide gels and related procedures. In: The proteins, vol. 1. edn. 3, pp. 179–223, Neurath, H., Hill, R.L., eds. Academic Press, New York

    Google Scholar 

Download references

Author information

Authors and Affiliations

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

    C. Domoney & R. Casey

Authors
  1. C. Domoney
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R. Casey
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Domoney, C., Casey, R. Cloning and characterization of complementary DNA for convicilin, a major seed storage protein in Pisum sativum L.. Planta 159, 446–453 (1983). https://doi.org/10.1007/BF00392081

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00392081

Key words

Search

Navigation