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Nif-hybrids of enterobacter: Selection for nif gene integration with chlorate

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Summary

The nif gene group from Klebsiella can be transferred into Enterobacter cloacae by conjugation using Escherichia coli donor cells carrying the composite self-transmissible nif-plasmid pRD1. A small fraction of the hybrids obtained is stable upon prolonged passaging without selection. Their stability is due to integration of pRD1 into the chromosome. Such integration hybrids were chlorate resistant, and nitrate reductase negative, which indicated that integration preferentially occurred within one of the genes for the production or functioning of this enzyme. Chlorate resistance could, therefore, be used to select for additional nitrate reductase-negative sublines with pRD1 in their chromosome. Such sublines have been analyzed further for the presence of nif genes, other pRD1 markers, and for stability. In all except one the complete plasmid seems to have been integrated. Some tend to revert to nitrate utilisation (chlorate sensitivity).

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References

  • Dixon RA, Cannon FC, Kondorosi A (1976) Construction of a P plasmid carrying nitrogen fixation genes from Klebsiella pneumoniae. Nature (London) 260:268–271

    Google Scholar 

  • Edwards PR, Ewing WH (1972) Identification of Enterobacteriaceae. Burgess Publ Co, Minneapolis, Minnesota

    Google Scholar 

  • Kado CI, Liu ST (1981) Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 145:1365–1373

    Google Scholar 

  • Kleeberger A, Klingmüller W (1980) Plasmid-mediated transfer of nitrogen fixing capability to bacteria from the rhizosphere of grasses. Mol Gen Genet 180:621–627

    Google Scholar 

  • Klingmüller W (1979) Genetic engineering for practical application. Naturwissenschaften 66:182–189

    Google Scholar 

  • Klingmüller W, Kleeberger A (1981) Übertragung der genetischen Information für Stickstoffbindung aus E. coli auf Bodenbakterien. Ber Dtsch Bot Ges 94:85–92

    Google Scholar 

  • Knowles R (1982) Denitrification. Microbiol Rev 46:43–70

    Google Scholar 

  • Lowe RH, Evans HJ (1964) Preparation and some properties of a soluble nitrate reductase from Rhizobium japonicum. Biochim Biophys Acta 85:377–389

    Google Scholar 

  • Pühler A, Burkardt HJ, Cannon FC, Wohlleben W (1979a) Spontaneous degradation of pRD1 DNA into unique size classes is recA dependent. Mol Gen Genet 171:1–6

    Google Scholar 

  • Pühler A, Burkardt HJ, Klipp W (1979b) Cloning of the entire region for nitrogen fixation from Klebsiella pneumoniae on a multicopy plasmid vehicle in Escherichia coli. Mol Gen Genet 176:17–24

    Google Scholar 

  • Singh M, Kleeberger A, Klingmüller W (1983) Location of nitrogen fixation (nif) genes on indigenous plasmids of Enterobacter agglomerans. Mol Gen Genet 190:373–378

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Van't Reit J, Stouthamer AH, Planta RJ (1968) Regulation of nitrate assimilation and nitrate respiration in Aerobacter aerogenes. J Bacteriol 96:1455–1464

    Google Scholar 

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Communicated by H. Böhme

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Klingmüller, W., Shanmugam, K.T. & Singh, M. Nif-hybrids of enterobacter: Selection for nif gene integration with chlorate. Mol Gen Genet 191, 221–224 (1983). https://doi.org/10.1007/BF00334817

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  • DOI: https://doi.org/10.1007/BF00334817

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