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Molecular cloning of lupin leghemoglobin cDNA

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Abstract

Poly(A)+RNA isolated from root nodules of yellow lupin (Lupinus luteus, var. Ventus) has been used as a template for the construction of a cDNA library. The ds cDNA was synthesized and inserted into the Hind III site of plasmid pBR 322 using synthetic Hind III linkers. Clones containing sequences specific for nodules were selected by differential colony hybridization using32P-labeled cDNA synthesized either from nodule poly(A)+RNA or from poly(A)+RNA of uninfected root as probes. Among the recombinant plasmids, the cDNA gene for leghemoglobin was identified. The protein structure derived from its nucleotide sequence was consistent with known amino acid sequence of lupin Lb II. The cloned lupin Lb cDNA hybridized to poly(A)+RNA from nodules only, which is in accordance with the general concept, that leghemoglobin is expressed exclusively in nodules.

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Authors and Affiliations

  1. Department of Biochemistry, University of Agriculture, Wołyńska 35, 60-637, Poznań, Poland

    A. Konieczny & A. B. Legocki

  2. Institute of Molecular Biology and Plant Physiology, Aarhus University, C. F. Mollers Alle 130, Aarhus, Denmark

    E. Ø. Jensen & K. A. Marcker

Authors
  1. A. Konieczny
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  2. E. Ø. Jensen
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  3. K. A. Marcker
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  4. A. B. Legocki
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Konieczny, A., Jensen, E.Ø., Marcker, K.A. et al. Molecular cloning of lupin leghemoglobin cDNA. Mol Biol Rep 12, 61–66 (1987). https://doi.org/10.1007/BF00580652

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  • DOI: https://doi.org/10.1007/BF00580652

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