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. 1982 Jul 24;10(14):4147–4157. doi: 10.1093/nar/10.14.4147

On the operon structure of the nitrogenase genes of Rhizobium leguminosarum and Azotobacter vinelandii.

A J Krol, J G Hontelez, B Roozendaal, A van Kammen
PMCID: PMC320789  PMID: 6289264

Abstract

The transcription of the nitrogenase genes in Rhizobium leguminosarum was studied by analysing total cellular RNA from bacteroids for the presence of nitrogenase messenger RNA. The RNA was separated by agarose gel electrophoresis and blotted onto nitrocellulose filters. Messenger RNA for nitrogenase was detected by hybridization with probes derived from plasmid pSA30, a recombinant plasmid carrying the nitrogenase genes of Klebsiella pneumoniae. In the same way nitrogenase mRNA was detected in RNA isolated from Azotobacter vinelandii and from Klebsiella pneumoniae, but only if both were cultured under conditions of derepression of the nitrogenase genes. The size of the RNA hybridizing with the probes indicates that in all three organisms the genes for the subunits of the two components of nitrogenase constitute a single operon.

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Selected References

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

  1. Bisseling T., van Staveren W., van Kammen A. The effect of waterlogging on the synthesis of the nitrogenase components in bacteroids of Rhizobium leguminosarum in root nodules of Pisum sativum. Biochem Biophys Res Commun. 1980 Apr 14;93(3):687–693. doi: 10.1016/0006-291x(80)91132-8. [DOI] [PubMed] [Google Scholar]
  2. Bisseling T., van den Bos R. C., van Kammen A. The effect of ammonium nitrate on the synthesis of nitrogenase and the concentration of leghemoglobin in pea root nodules induced by Rhizobium leguminosarum. Biochim Biophys Acta. 1978 Feb 13;539(1):1–11. doi: 10.1016/0304-4165(78)90115-0. [DOI] [PubMed] [Google Scholar]
  3. Bánfalvi Z., Sakanyan V., Koncz C., Kiss A., Dusha I., Kondorosi A. Location of nodulation and nitrogen fixation genes on a high molecular weight plasmid of R. meliloti. Mol Gen Genet. 1981;184(2):318–325. doi: 10.1007/BF00272925. [DOI] [PubMed] [Google Scholar]
  4. 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]
  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. 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]
  7. Krol A. J., Hontelez J. G., Van den Bos R. C., Van Kammen A. Expression of large plasmids in the endosymbiotic form of Rhizobium leguminosarum. Nucleic Acids Res. 1980 Oct 10;8(19):4337–4347. doi: 10.1093/nar/8.19.4337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Merrick M., Filser M., Kennedy C., Dixon R. Polarity of mutations induced by insertion of transposons Tn5, Tn7 and Tn10 into the nif gene cluster of Klebsiella pneumoniae. Mol Gen Genet. 1978 Sep 20;165(1):103–111. doi: 10.1007/BF00270382. [DOI] [PubMed] [Google Scholar]
  11. Prakash R. K., Schilperoort R. A., Nuti M. P. Large plasmids of fast-growing rhizobia: homology studies and location of structural nitrogen fixation (nif) genes. J Bacteriol. 1981 Mar;145(3):1129–1136. doi: 10.1128/jb.145.3.1129-1136.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  13. Rosenberg C., Boistard P., Dénarié J., Casse-Delbart F. Genes controlling early and late functions in symbiosis are located on a megaplasmid in Rhizobium meliloti. Mol Gen Genet. 1981;184(2):326–333. doi: 10.1007/BF00272926. [DOI] [PubMed] [Google Scholar]
  14. Ruvkun G. B., Ausubel F. M. A general method for site-directed mutagenesis in prokaryotes. Nature. 1981 Jan 1;289(5793):85–88. doi: 10.1038/289085a0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Sanders J. P., Borst P., Weijers P. J. The organization of genes in yeast mitochondrial DNA II. The physical map of EcoRI and HindII + III fragments. Mol Gen Genet. 1975 Dec 30;143(1):53–64. doi: 10.1007/BF00269420. [DOI] [PubMed] [Google Scholar]
  17. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Thuring R. W., Sanders J. P., Borst P. A freeze-squeeze method for recovering long DNA from agarose gels. Anal Biochem. 1975 May 26;66(1):213–220. doi: 10.1016/0003-2697(75)90739-3. [DOI] [PubMed] [Google Scholar]

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