Skip to main content
SpringerLink
Log in

Molecular cloning of a pea mRNA encoding an early light induced, nuclear coded chloroplast protein

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Summary

cDNA clones were isolated for a chloroplast protein, the mRNA of which is induced to maximum levels within 2–4 h after onset of illumination in five day old, etiolated pea seedlings.

The cDNA library was constructed from poly(A)+-mRNA which was isolated from 4 h illuminated seedlings. The extremely short induction period of the early light induced protein(ELIP)-mRNA established the basis of our screening procedure. Colony hybridization experiments were performed with32P-labelled cDNA probes, synthesized from RNA of seedlings which had been exposed to different programs of illumination. Plasmid DNAs were isolated from colonies showing strong hybridization signals exclusively with cDNA corresponding to the 4 h-mRNA. Hybrid released translation of preselected plasmids p 17/C2 and p17/C4 revealed a peptide of Mr 24 000. After posttranslational importin vitro, the processed product of Mr 17 000 appears in the chloroplast. Using these clones, the expression of the ELIP-mRNA was investigated by DOT-hybridization. The ELIP-mRNA reaches maximum levels within 2–4 hours after onset of illumination. Our results correspond precisely to thein vivo characteristics and indicate positive identification of the sought clones.

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

References

  1. Apel K, Kloppstech K: The plastid membranes of barley (Hordeum vulgare). Light-induced appearance of mRNAs coding for the apo-protein of the light-harvesting chlorophyll a/b protein. Eur J Biochem 85: 581–588, 1978.

    Article  CAS  PubMed  Google Scholar 

  2. Batschauer A, Apel K: An inverse control by phytochrome of the expression of two nuclear genes in barley (Hordeum vulgare L.). Eur J Biochem 143: 593–597, 1984.

    Article  CAS  PubMed  Google Scholar 

  3. Bedbrook JR, Smith SM, Ellis RJ: Molecular cloning and sequencing of cDNA encoding the precursor to the small subunit of chloroplast RuBPCase. Nature 287: 692–697, 1980.

    Article  CAS  Google Scholar 

  4. Birnboim HC, Doly J: A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7: 1513, 1979.

    CAS  PubMed  Google Scholar 

  5. Broglie R, Bellemare G, Bartlett SG, Chua NH, Cashmore AR: Cloned DNA sequences complementary to mRNAs encoding precursors to the small subunit of ribulose-1,5-bisphosphate carboxylase and a chlorophyll binding polypeptide. Proc Natl Acad Sci 78:7304–7308, 1981.

    CAS  PubMed  Google Scholar 

  6. Domoney C, Casey R: Storage protein precursor polypeptides in cotyledons of Pisum sativum L. Identification of, and isolation of a cDNA clone for, an 80 000 Mr legumin related polypeptide. Eur J Biochem 139: 321–327, 1984.

    Article  CAS  PubMed  Google Scholar 

  7. Grunstein M, Hogness DS: Colony hybridization: A method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci 72:3961–3965, 1975.

    CAS  PubMed  Google Scholar 

  8. Hampp R, de Filippis LF: Plastid protease activity and prolamellar body transformation during greening. Plant Physiol 65: 663–668, 1980.

    CAS  PubMed  Google Scholar 

  9. Hanahan D: Studies on Transformation of Escherichia coli with Plasmids. J Mol Biol 166: 557–580, 1983.

    CAS  PubMed  Google Scholar 

  10. Maniatis T, Fritsch EF, Sambrook J: Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, New York, 1982.

    Google Scholar 

  11. Mc Master GK, Carmichael GC: Analysis of single and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridin orange. Proc Natl Acad Sci 74: 4835–4838, 1977.

    CAS  Google Scholar 

  12. Meyer G, Kloppstech K: A rapidly light-induced chloroplast protein with a high turnover coded for by pea nuclear DNA. Eur J Biochem 138: 201–207, 1984.

    Article  CAS  PubMed  Google Scholar 

  13. Ricciardi RP, Miller JS, Roberts BE: Purification and mapping of specific mRNAs by hybridization selection and cell-free translation. Proc Natl Acad Sci 76: 4927–4931, 1979.

    CAS  PubMed  Google Scholar 

  14. Rigby PWJ, Dieckmann M, Rhodes C, Berg P: Labelling deoxyribonucleic acid to high specific activityin vitro by nick translation with DNA polymerase I. J Mol Biol 113: 237, 1977.

    Article  CAS  PubMed  Google Scholar 

  15. Roychoudhoury R, Jay E, Wu R: Terminal labelling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase. Nucleic Acids Res 3: 863–877, 1976.

    Google Scholar 

  16. Stiekema WJ, Wimpee CF, Silverthorne J, Tobin E: Phyochrome control of the Expression of Two Nuclear Genes Encoding Chloroplast Proteins in Lemna gibba L. Plant Physiol 72: 717–724, 1983.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Botanik, Fachbereich Biologie, Universität Hannover, Herrenhäuser Str. 2, 3000, Hannover, F. R. G.

    Christina Scharnhorst, Hartmut Heinze, Gabriele Meyer, Waldemar Kolanus, Klaus Bartsch, Susanne Heinrichs, Thomas Gudschun, Margret Möller & Frank Herzfeld

Authors
  1. Christina Scharnhorst
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hartmut Heinze
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gabriele Meyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Waldemar Kolanus
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Klaus Bartsch
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Susanne Heinrichs
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Thomas Gudschun
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Margret Möller
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Frank Herzfeld
    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

Scharnhorst, C., Heinze, H., Meyer, G. et al. Molecular cloning of a pea mRNA encoding an early light induced, nuclear coded chloroplast protein. Plant Mol Biol 4, 241–245 (1985). https://doi.org/10.1007/BF02418242

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02418242

Keywords

Search

Navigation