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Isolation and organization of genes for nitrogen fixation in Rhodopseudomonas capsulata

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Summary

A library of Rhodopseudomonas capsulata chromosomal DNA was constructed in the broad host range cosmid vector pLAFR1. The library was used to isolate nitrogen fixation genes by complementation of R. capsulata Nif- mutants. Four complementing regions were localized on different cloned DNA fragments by Tn5 and mini-Mu mutagenesis. Additional nif genes were identified by recombination of transposons from the nif cosmids into the R. capsulata chromosome resulting in the creation of new Nif- mutations. Most of the newly cloned DNA fragments containing nif genes were found to be unlinked to any other by Southern hybridization of the cloned DNA to chromosomal DNA blots. One of the new fragments was linked to the nifHDK genes. Another cluster spanning 10–12 kilobase pairs contained a number of nif genes, possibly as many as eight.

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References

  • Ausubel FM, Cannon FC (1980) Molecular genetic analysis of Klebsiella pneumoniae nitrogen fixation (nif) genes. Cold Spring Harbor Symp Quant Biol 45:487–497

    Google Scholar 

  • Avtges P, Scolnik PA, Haselkorn R (1983) Genetic and physical map of the structural genes (nifHDK) coding for the nitrogenase complex of Rhodopseudomonas capsulata. J Bacteriol 156:251–256

    Google Scholar 

  • Berg DE, Weiss A, Crossland L (1980) Polarity of Tn5 insertions in Escherichia coli. J Bacteriol 142:439–446

    Google Scholar 

  • Boyer HW, Rouland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472

    Google Scholar 

  • Brill W (1980) Biochemical genetics of nitrogen fixation. Microbiol Rev 44:449–467

    Google Scholar 

  • Castilho BA, Olfson P, Casadaban MT (1984) Plasmid insertion mutagenesis and lac gene fusion with mini-Mu bacteriophage transposons. J Bacteriol 158:488–495

    Google Scholar 

  • Ditta G, Stanfield S, Corbin D, Helinski D (1980) Broad host range cloning system for gram negative bacteria: contruction of a gene bank of Rhizobium meliloti. Proc Natl Acac Sci USA 77:7347–7354

    Google Scholar 

  • Enquist L, Sternberg N (1979) In vitro packaging of λdam vectors and their use in cloning DNA fragments. Meth Enzymol 68:281–298

    Google Scholar 

  • Friedman AM, Long SR, Brown SE, Buikema WJ, Ausubel FM (1982) Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene 18:289–296

    Google Scholar 

  • Hirsch PR, Beringer JE (1984) A physical map of pPH1J1 and pJB4J1. Plasmid 12:139–141

    Google Scholar 

  • Johansson BC, Gest H (1977) Adenylation/deadenylation control of the glutamine synthetase of Rhodopseudomonas capsulata. Eur J Biochem 81:365–371

    Google Scholar 

  • Kranz R, Haselkorn R (1985) Characterization of nif regulatory genes in Rhodopseudomonas capsulata using lac fusions. Gene, in press

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, p 433

    Google Scholar 

  • Pühler A, Aguilar MO, Hynes M, Müller P, Klipp W, Priefer U, Simon R, Weber G (1984) Advances in the genetics of free-living and symbiotic nitrogen fixing bacteria. In: Veeger C, Newton WE (eds) Advances in nitrogen fixation research, Nijhoff/Junk, The Hague, pp 609–619

    Google Scholar 

  • Ruvkun GB, Ausubel FM (1981) A general method for site-directed mutagenesis in prokaryotes. Nature 289:85–89

    Google Scholar 

  • Scolnik PA, Haselkorn R (1984) Activation of extra copies of genes coding for nitrogenase in Rhodopseudomonas capsulata. Nature 307:289–292

    Google Scholar 

  • Scolnik PA, Virosco J, Haselkorn R (1983) The wild type gene for glutamine synthetase restores ammonia control of nitrogen fixation to Gln-(glnA) mutants of Rhodopseudomonas capsulata. J Bacteriol 155:180–185

    Google Scholar 

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

    Google Scholar 

  • Wall JD, Braddock K (1984) Genetic mapping of nif genes in Rhodopseudomonas capsulata. J Bacteriol 158:404–410

    Google Scholar 

  • Wall JD, Love J, Quinn SP (1984) Spontaneous Nif- mutants of Rhodopseudomonas capsulata. J Bacteriol 159:652–657

    Google Scholar 

  • Yen HC, Marrs B (1976) Map of genes of carotenoid and bacteriochlorophyll biosyntheesis in Rhodopseudomonas capsulata. J Bacteriol 126:619–629

    Google Scholar 

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Authors and Affiliations

  1. Department of Molecular Genetics and Cell Biology, The University of Chicago, 60637, Chicago, IL, USA

    Peter Avtges, Robert G. Kranz & Robert Haselkorn

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  1. Peter Avtges
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  2. Robert G. Kranz
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  3. Robert Haselkorn
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Communicated by P. Tiollais

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Avtges, P., Kranz, R.G. & Haselkorn, R. Isolation and organization of genes for nitrogen fixation in Rhodopseudomonas capsulata . Molec Gen Genet 201, 363–369 (1985). https://doi.org/10.1007/BF00331324

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  • DOI: https://doi.org/10.1007/BF00331324

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