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A System of Oligonucleotide Primers for the Amplification of nifH Genes of Different Taxonomic Groups of Prokaryotes

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Abstract

Based on the analysis of the nifH gene nucleotide sequences from GenBank, a system of primers was developed that makes it possible to obtain 370- and 470-bp PCR fragments of the nifH gene of nitrogen-fixing bacteria and archaea. The effectiveness of the proposed system for revealing the presence of nifH genes was demonstrated by PCR on the DNA isolated from nitrogen-fixing prokaryotes for which the primary structure of these genes is known and which belong to different taxonomic groups. nifH sequences of nitrogen-fixing prokaryotes of the genera Xanthobacter, Beijerinckia, and Methanosarcina, for which the capacity for nitrogen fixation was demonstrated earlier, but no data existed on the nucleotide composition of these genes, were determined and deposited in GenBank.

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References

  1. Stackebrandt, E. and Goebel, B.M., Taxonomic Note: A Place for DNA–DNA Reassociation and 16S rRNA Sequence Analysis in the Present Species Definition in Bacteriology, Int. J. Syst. Bacteriol., 1994, vol. 44, pp. 846–849.

    Google Scholar 

  2. Young, J.P.W., Phylogenetic Classification of Nitrogen-Fixing Organisms, Biological Nitrogen Fixation, Stacey, G., Burris, R.H., and Evans, H.G., Eds., New York: Chapman and Hall, 1992, pp. 43–86.

    Google Scholar 

  3. Ueda, T., Suga, Y., Yashiro, N., and Matsuguchi, T., Remarkable N2-Fixing Bacterial Diversity Detected in Rice Roots by Molecular Evolutionary Analysis of nifH Gene Sequences, J. Bacteriol., 1995, vol. 177, pp. 1414–1417.

    Google Scholar 

  4. Zehr, J.P., Mellon, M., Braun, S., Litaker, W., Steppe, T., and Paerl, H.W., Diversity of Heterotrophic Nitrogen Fixation Genes in a Marine Cyanobacterial Mat, Appl. Environ. Microbiol., 1995, vol. 61, pp. 2527–2532.

    Google Scholar 

  5. Ben-Porath, J. and Zehr, J.P., Detection and Characterization of Cyanobacterial nifH Genes, Appl. Environ. Microbiol., 1995, vol. 60, pp. 880–887.

    Google Scholar 

  6. Okhuma, M., Noda, S., Usami, R., Horikoshi, K., and Kudo, T., Diversity of Nitrogen Fixation Genes in the Symbiotic Intestinal Microflora of the Termite Reticulitermis speratus, Appl. Environ. Microbiol., 1996, vol. 62, pp. 2747–2752.

    Google Scholar 

  7. Kirshtein, J.D., Paerl, H.W., and Zehr, J., Amplification, Cloning, and Sequencing of a nifH Segment from Aquatic Microorganisms and Natural Communities, Appl. Environ. Microbiol., 1991, vol. 54, pp. 2645–2650.

    Google Scholar 

  8. Zehr, J. and McReynolds, L., Use of Degenerate Oligonucleotide Primers for Amplification of the nifH Gene from the Marine Cyanobacterium Trihodesmium thiebautii, Appl. Environ. Microbiol., 1989, vol. 55, pp. 2427–2532.

    Google Scholar 

  9. Ohkuma, M., Noda, S., and Kudo, T., Phylogenetic Diversity of Nitrogen Fixation Genes in the Symbiotic Microbial Community in the Gut of Diverse Termites, Appl. Environ. Microbiol., 1999, vol. 65, pp. 4926–4934.

    Google Scholar 

  10. Widmer, F., Shaffer, B.T., Porteous, L.A., and Seidler, R.J., Analysis of nifH Gene Pool Complexity in Soil and Litter at a Douglas Fir Forest Site in the Oregon Cascade Mountain Range, Appl. Environ. Microbiol., 1999, vol. 65, pp. 374–380.

    Google Scholar 

  11. Balows, A., Trüper, H.G., Dworkin, M., Harder, W., and Schleifer, K.-H., The Prokaryotes, New York: Springer-Verlag, 1992, vol. 2, pp. 2065–2066.

    Google Scholar 

  12. Jarvis, B.D.W., Gillis, M., and De Ley, J., Rhizobium loti, a New Species of Legume Root Nodule Bacteria, Int. J. Syst. Bacteriol., 1982, vol. 32, pp. 378–380.

    Google Scholar 

  13. Metody pochvennoi mikrobiologii i biokhimii (Methods in Soil Microbiology and Biochemistry), Zvyagintsev, D.G., Ed., Moscow: Mosk. Gos. Univ., 1991, p. 304.

    Google Scholar 

  14. Sanger, F., Nicklen, S., and Coulson, A.R., DNA Sequencing with Chain-Terminating Inhibitors, Proc. Natl. Acad. Sci. USA, 1977, vol. 84, pp. 5463–5467.

    Google Scholar 

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Marusina, A.I., Boulygina, E.S., Kuznetsov, B.B. et al. A System of Oligonucleotide Primers for the Amplification of nifH Genes of Different Taxonomic Groups of Prokaryotes. Microbiology 70, 73–78 (2001). https://doi.org/10.1023/A:1004849022417

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  • DOI: https://doi.org/10.1023/A:1004849022417

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