Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1980 Jan;77(1):186–190. doi: 10.1073/pnas.77.1.186

Identification of blue-green algal nitrogen fixation genes by using heterologous DNA hybridization probes

Barbara J Mazur 1,*, Douglas Rice 1, Robert Haselkorn 1
PMCID: PMC348233  PMID: 16592748

Abstract

In the filamentous blue-green alga Anabaena 7120, aerobic nitrogen fixation is linked to the differentiation of specialized cells called heterocysts. In order to study control of heterocyst development and nitrogen fixation in Anabaena, we have used cloned fragments of the Klebsiella pneumoniae nitrogen fixation (nif) genes as probes in DNA·DNA hybridizations with restriction endonuclease fragments of Anabaena DNA. Using this technique, we were able to identify and clone Anabaena nif genes, demonstrating the feasibility of using heterologous probes to identify genes for which no traditional genetic selection exists. From the patterns of hybridization observed, we deduced that although DNA sequence homology has been retained between some of the nif genes of these divergent organisms, the nif gene order has been rearranged.

Keywords: Klebsiella pneumoniae DNA probes, recombinant DNA, Anabaena

Full text

PDF
187

Images in this article

187Image
on p.187
188Image
on p.188
188Image
on p.188
189Image
on p.189

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ausubel F., Riedel G., Cannon F., Peskin A., Margolskee R. Cloning nitrogen fixing genes from Klebsiella pneumoniae in vitro and the isolation of NIF promoter mutants affecting glutamine synthetase regulation. Basic Life Sci. 1977;9:111–128. doi: 10.1007/978-1-4684-0880-5_10. [DOI] [PubMed] [Google Scholar]
  2. Cannon F. C., Riedel G. E., Ausubel F. M. Overlapping sequences of Klebsiella pneumoniae nifDNA cloned and characterized. Mol Gen Genet. 1979 Jul 2;174(1):59–66. doi: 10.1007/BF00433306. [DOI] [PubMed] [Google Scholar]
  3. Cannon F. C., Riedel G. E., Ausubel F. M. Recombinant plasmid that carries part of the nitrogen fixation (nif) gene cluster of Klebsiella pneumoniae. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2963–2967. doi: 10.1073/pnas.74.7.2963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Currier T. C., Haury J. F., Wolk C. P. Isolation and preliminary characterization of auxotrophs of a filamentous Cyanobacterium. J Bacteriol. 1977 Mar;129(3):1556–1562. doi: 10.1128/jb.129.3.1556-1562.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  6. Dixon R., Kennedy C., Kondorosi A., Krishnapillai V., Merrick M. Complementation analysis of Klebsiella pneumoniae mutants defective in nitrogen fixation. Mol Gen Genet. 1977 Nov 29;157(2):189–198. doi: 10.1007/BF00267397. [DOI] [PubMed] [Google Scholar]
  7. Elmerich C., Houmard J., Sibold L., Manheimer I., Charpin N. Genetic and biochemical analysis of mutants induced by bacteriophage Mu DNA integration into Klebsiella pneumoniae nitrogen fixation genes. Mol Gen Genet. 1978 Oct 4;165(2):181–189. doi: 10.1007/BF00269905. [DOI] [PubMed] [Google Scholar]
  8. Elmerich C., Houmard J., Sibold L., Manheimer I., Charpin N. Genetic and biochemical analysis of mutants induced by bacteriophage Mu DNA integration into Klebsiella pneumoniae nitrogen fixation genes. Mol Gen Genet. 1978 Oct 4;165(2):181–189. doi: 10.1007/BF00269905. [DOI] [PubMed] [Google Scholar]
  9. Fay P., Stewart W. D., Walsby A. E., Fogg G. E. Is the heterocyst the site of nitrogen fixation in blue-green algae? Nature. 1968 Nov 23;220(5169):810–812. doi: 10.1038/220810b0. [DOI] [PubMed] [Google Scholar]
  10. MacNeil T., MacNeil D., Roberts G. P., Supiano M. A., Brill W. J. Fine-structure mapping and complementation analysis of nif (nitrogen fixation) genes in Klebsiella pneumoniae. J Bacteriol. 1978 Oct;136(1):253–266. doi: 10.1128/jb.136.1.253-266.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mazur B. J., Model P. Regulation of coliphage f1 single-stranded DNA synthesis by a DNA-binding protein. J Mol Biol. 1973 Aug 5;78(2):285–300. doi: 10.1016/0022-2836(73)90117-4. [DOI] [PubMed] [Google Scholar]
  13. Model P., Zinder N. D. In vitro synthesis of bacteriophage f1 proteins. J Mol Biol. 1974 Feb 25;83(2):231–251. doi: 10.1016/0022-2836(74)90389-1. [DOI] [PubMed] [Google Scholar]
  14. Nagatani H. H., Haselkorn R. Molybdenum independence of nitrogenase component synthesis in the non-heterocystous cyanobacterium Plectonema. J Bacteriol. 1978 May;134(2):597–605. doi: 10.1128/jb.134.2.597-605.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Riedel G. E., Ausubel F. M., Cannon F. C. Physical map of chromosomal nitrogen fixation (nif) genes of Klebsiella pneumoniae. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2866–2870. doi: 10.1073/pnas.76.6.2866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Roberts G. P., MacNeil T., MacNeil D., Brill W. J. Regulation and characterization of protein products coded by the nif (nitrogen fixation) genes of Klebsiella pneumoniae. J Bacteriol. 1978 Oct;136(1):267–279. doi: 10.1128/jb.136.1.267-279.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ruvkun G. B., Ausubel F. M. Interspecies homology of nitrogenase genes. Proc Natl Acad Sci U S A. 1980 Jan;77(1):191–195. doi: 10.1073/pnas.77.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
  19. Simon R. D. Survey of extrachromosomal DNA found in the filamentous cyanobacteria. J Bacteriol. 1978 Oct;136(1):414–418. doi: 10.1128/jb.136.1.414-418.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  21. St John R. T., Johnston H. M., Seidman C., Garfinkel D., Gordon J. K., Shah V. K., Brill W. J. Biochemistry and genetics of Klebsiella pneumoniae mutant strains unable to fix N2. J Bacteriol. 1975 Mar;121(3):759–765. doi: 10.1128/jb.121.3.759-765.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stewart W. D., Lex M. Nitrogenase activity in the blue-green alga Plectonema boryanum strain 594. Arch Mikrobiol. 1970;73(3):250–260. doi: 10.1007/BF00410626. [DOI] [PubMed] [Google Scholar]
  23. Streicher S. L., Shanmugam K. T., Ausubel F., Morandi C., Goldberg R. B. Regulation of nitrogen fixation in Klebsiella pneumoniae: evidence for a role of glutamine synthetase as a regulator of nitrogenase synthesis. J Bacteriol. 1974 Nov;120(2):815–821. doi: 10.1128/jb.120.2.815-821.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tsai L. B., Mortenson L. E. Interaction of the nitrogenase components of Anabaena cylindrica with those of Clostridium pasteurianum. Biochem Biophys Res Commun. 1978 Mar 30;81(2):280–287. doi: 10.1016/0006-291x(78)91530-9. [DOI] [PubMed] [Google Scholar]
  25. Tubb R. S. Glutamine synthetase and ammonium regulation of nitrogenase synthesis in Klebsiella. Nature. 1974 Oct 11;251(5475):481–485. doi: 10.1038/251481a0. [DOI] [PubMed] [Google Scholar]
  26. Wilcox M., Mitchison G. J., Smith R. J. Mutants of Anabaena cylindrica altered in heterocyst spacing. Arch Microbiol. 1975 May 5;103(3):219–223. doi: 10.1007/BF00436353. [DOI] [PubMed] [Google Scholar]
  27. Wolk C. P. Physiological basis of the pattern of vegetative growth of a blue-green alga. Proc Natl Acad Sci U S A. 1967 May;57(5):1246–1251. doi: 10.1073/pnas.57.5.1246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wolk C. P., Shaffer P. W. Heterotrophic micro- and macrocultures of a nitrogen-fixing cyanobacterium. Arch Microbiol. 1976 Nov 2;110(23):145–147. doi: 10.1007/BF00690221. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES