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Characterization and nucleotide sequence of a novel gene fixW upstream of the fixABC operon in Rhizobium leguminosarum

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Summary

On the Rhizobium leguminosarum PRE sym plasmid, fixABC and a novel gene fixW were identified upstream of the regulatory gene nifA. The molecular masses of FixABC, 29, 44 and 50 kDa respectively, were estimated by polyacrylamide gel electrophoresis (PAGE) and of FixW, 25 kDa, by PAGE and nucleotide sequencing. Hybridization studies using bacteroid mRNA as a probe showed that fixABC is one operon which can be transcribed independently of fixW. Nucleotide sequencing revealed that both fixW and fixA are preceded by a nif consensus promoter. The fixA promoter partly overlaps the 3′-terminal coding region of fixW, indicating that readthrough from fixW into fixA is possible. Two open reading frames, ORF71 and ORF79, precede fixW and form one operon with fixW. ORF71 contains sequences homologous to the fixA promoter and 5′-terminal coding region. One more duplication of fixA sequences was detected, also located within the sym plasmid nif/fix clusters. One duplication of fixW sequences was found. No fixW homologue could be found in other nitrogen fixing organisms except in a number of R. leguminosarum strains.

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Authors and Affiliations

  1. Department of Molecular Biology, Agricultural University, Dreijenlaan 3, NL-6703 HA, Wageningen, The Netherlands

    Jan G. J. Hontelez, René Klein Lankhorst, Rommert C. van den Bos & Albert van Kammen

  2. School of Science, Department of Biology, Aristotelian University of Thessaloniki, 54006, Thessaloniki, Greece

    Panagiotis Katinakis

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  1. Jan G. J. Hontelez
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  2. René Klein Lankhorst
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  3. Panagiotis Katinakis
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  4. Rommert C. van den Bos
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  5. Albert van Kammen
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Communicated by H. Henneke

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Hontelez, J.G.J., Lankhorst, R.K., Katinakis, P. et al. Characterization and nucleotide sequence of a novel gene fixW upstream of the fixABC operon in Rhizobium leguminosarum . Mol Gen Genet 218, 536–544 (1989). https://doi.org/10.1007/BF00332421

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  • DOI: https://doi.org/10.1007/BF00332421

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