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Sequence analysis of pre-ferredoxin-NADP+-reductase cDNA from Cyanophora paradoxa specifying a precursor for a nucleus-encoded cyanelle polypeptide

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Abstract

A cDNA clone for pre-ferredoxin-NADP+ reductase (FNR) was obtained by screening a Cyanophora paradoxa expression library with antibodies specific for cyanelle FNR. The 1.4 kb transcript was derived from a single-copy gene. The precursor (41 kDa) and mature forms (34 kDa) of FNR were identified by western blotting of in vitro translation products and cyanelle extracts, respectively. The derived amino acid sequence of the mature form was corroborated by data from N-terminal protein sequencing and yielded identity scores from 58% to 62% upon comparison with cyanobacterial FNRs. Sequence conservation seemed to be even more pronounced in comparison with enzymes from higher plants, but using the neighbor joining method the C. paradoxa sequence was clearly positioned between the prokaryotic and eukaryotic sequences. The transit peptide of 65 or 66 amino acids appeared to be totally unrelated to those from spinach, pea and ice plant but showed overall characteristics of stroma-targeting peptides.

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Authors and Affiliations

  1. Institut für Biochemie und Molekulare Zellbiologie der Universität Wien und Ludwig Boltzmann-Forschungsstelle für Biochemie, A-1030, Vienna, Austria

    Johannes Jakowitsch, Martin Brandtner, Barbara Hamilton, Christoph Neumann-Spallart & Wolfgang Löffelhardt

  2. Botanisches Institut, Lehrstuhl für Allgemeine Botanik und Pflanzenphysiologie, Universität Tübingen, D-7400, Tübingen, Germany

    Manfred G. Bayer, Thomas L. Maier, Ulrike B. Gebhart & Hainfried E. A. Schenk

  3. Arnimstr. 2, D-5000, Köln, Germany

    Angela Lüttke

  4. Department of Biochemistry, University of Arizona, 85 721, Tucson, AZ, USA

    Christine B. Michalowski & Hans J. Bohnert

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  1. Johannes Jakowitsch
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  2. Manfred G. Bayer
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  3. Thomas L. Maier
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  4. Angela Lüttke
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  5. Ulrike B. Gebhart
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  6. Martin Brandtner
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  7. Barbara Hamilton
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  8. Christoph Neumann-Spallart
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  9. Christine B. Michalowski
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  10. Hans J. Bohnert
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  11. Hainfried E. A. Schenk
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  12. Wolfgang Löffelhardt
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Jakowitsch, J., Bayer, M.G., Maier, T.L. et al. Sequence analysis of pre-ferredoxin-NADP+-reductase cDNA from Cyanophora paradoxa specifying a precursor for a nucleus-encoded cyanelle polypeptide. Plant Mol Biol 21, 1023–1033 (1993). https://doi.org/10.1007/BF00023600

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  • DOI: https://doi.org/10.1007/BF00023600

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