Skip to main content
Log in

Overlapping sequences of Klebsiella pneumoniae nif DNA cloned and characterised

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

A HindIII (17.0 kb) and an EcoRl restriction fragment (6.9 kb) of Klebsiella pneumoniae nif DNA were cloned on two small amplifiable plasmids, pCM1 and pSA30 respectively. These plasmids between them carry 14 of the 15 known Klebsiella nif genes. The operon for the three structural genes for nitrogenase, nifpHDK, is carried on pSA30: four and five of the remaining six operons are on pCRA37 and pCM1 respectively. All of the nif genes were assigned to endonculease restriction fragments of DNA using the Southern blotting technique (Southern, 1975) with total DNA of nif insertion mutants and radioactive plasmid DNA which contained cloned nif DNA sequences. Their locations were consistent with the genetic map of nif genes. The estimated size of the nif gene cluster was 24 kb.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ausubel, F., Riedel, G., Cannon, F., Peskin, A., Margolskee, R.: Cloning nitrogen fixation genes from Klebsiella pneumoniae in vitro and the isolation of nif promoter mutants. In: Genetic engineering for nitrogen fixation (A. Hollander, ed.), pp. 111–128. New York: Plenum Press 1977

    Google Scholar 

  • Boyer, H.W., Betlach, M., Bolivar, F., Rodriguez, R.L., Heyneker, H.L., Shine, J., Goodman, H.M.: The construction of molecular cloning vehicles. In: Recombinant molecules: Impact on science and society (R.F. Beer and E.G. Bassett, eds.), pp. 9–20. New York: Raven Press 1977

    Google Scholar 

  • Cannon, F.: Cloning of the Klebsiella nitrogen fixation (nif) genes. In: Genetic engineering (H.W. Boyer and S. Nicosia, eds.), pp. 181–188. Amsterdam: Elsevier/North-Holland, Biochemical Press 1978

    Google Scholar 

  • Cannon, F.C., Dixon, R.A., Postgate, J.R.: Derivation and properties of F-prime factors in Escherichia coli carrying nitrogen fixation genes from Klebsiella pneumoniae. J. Gen. Microbiol. 93, 11–125 (1976)

    Google Scholar 

  • Cannon, F.C., Riedel, G.E., Ausubel, F.M.: Recombinant plasmid that carries part of the nitrogen fixation gene cluster of Klebsiella pneumoniae. Proc. Natl. Acad. Sci. U.S.A. 74, 2963–2967 (1977)

    Google Scholar 

  • Chang, A.C.Y., Cohen, S.N.: Construction and characterisation of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J. Bacteriol. 134, 1141–1156 (1978)

    Google Scholar 

  • Davis, B.D., Mingioli, E.S.: Mutants of Escherichia coli requiring methionine or vitamin B12. J. Bacteriol. 60, 17–28, (1950)

    Google Scholar 

  • Dixon, R.A., Cannon, F.C., Kondorosi, A.: Construction of a P plasmid carrying nitrogen fixation from Klebsiella pneumoniac. Nature 260, 268–271 (1976)

    Google Scholar 

  • Dixon, R.A., Kennedy, C., Kondorosi, A., Krisnapillai, V., Merrick, M.: Complementation analysis of Klebsiella pneumoniae mutants defective in nitrogen fixation. Mol. Gen. Genet. 157, 189–198 (1977)

    Google Scholar 

  • Eady, R.R., Smith, B.E., Cook, K.A., Postgate, J.R.: Nitrogenase of Klebsiella pneumoniae. Purification and properties of the component proteins. Biochem. J. 128, 655–675 (1972)

    Google Scholar 

  • Elmerich, C., Houmard, J., Sibold, L., Marheimer, I., Charpin, N.: Genetic and biochemical analysis of mutants induced by bacteriophage Mu DNA integration into Klebsiella pneumoniae nitrogen fixation genes. Mol. Gen. Genet. 165, 181–189 (1978)

    Google Scholar 

  • Holloway, B.S., Richmond, M.H.: R factors used for genetic studies in strains of Pseudomonas aeruginosa and their origin. Genet. Res. Camb. 21, 103–105 (1973)

    Google Scholar 

  • Humphreys, G.O., Willshaw, G.A., Anderson, E.S.: A simple method for the preparation of large quantities of pure plasmid DNA. Biochim. Biophys. Acta 282, 457–463 (1975)

    Google Scholar 

  • Janssen, K.A., Riedel, G.E., Ausubel, F.M., Cannon, F.C.: Transcriptional studies with cloned nitrogen fixing genes. In: Proceedings of the 3rd International Symposium on nitrogen fixation (W.H. Orme-Johnson and W. Newton, eds.) held in Madison, Wisconsin, U.S.A., May 1978

  • Kennedy, C.: Linkage map of the nitrogen fixation (nif) genes in Klebsiella pneumoniae. Mol. Gen. Genet. 157, 199–204 (1977)

    Google Scholar 

  • 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. 136, 253–266 (1978)

    Google Scholar 

  • Merrick, M., Filser, M., Dixon, R., Elmerich, C., Sibold, L., Houmard, J.: Fine structure mapping of insertion mutations in the nif gene cluster of Klebsiella pneumoniae (in preparation)

  • 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. 165, 103–111 (1978)

    Google Scholar 

  • Puhler, A., Bukardt, H.J., Klipp, W., Wohlleben, W.: Identification of the gene region for nitrogen fixation from Klebsiella pneumoniae on the P-type plasmid RP41. In: Plasmids and other extrachromosomal genetic elements (K. Timmis and W. Goebel, eds.), p. 81. Berlin: EMBO WORKSHOP Proceedings, Max Planck Institute for Molecular Genetics 1978

  • Riedel, G.E., Ausubel, F.M., Cannon, F.C.: Physically mapping mutations: the physical map of the nitrogen fixation (nif) genes of Klebsiella pneumoniae. Proc. Natl. Acad. Sci. U.S.A. (in press)

  • Roberts, G.P., MacNeil, T., MacNeil, D., Brill, W.J.: Regulation and characterisation of protein products coded by the nif (Nitrogen Fixation) genes in Klebsiella pneumoniae. J. Bacteriol. 136, 267–294 (1978)

    Google Scholar 

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

    Google Scholar 

  • Steicher, S.L., Gurney, E.G., Valentine, R.C.: The nitrogen fixation genes. Nature 239, 495–499 (1972)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by B.A. Bridges

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cannon, F.C., Riedel, G.E. & Ausubel, F.M. Overlapping sequences of Klebsiella pneumoniae nif DNA cloned and characterised. Molec. Gen. Genet. 174, 59–66 (1979). https://doi.org/10.1007/BF00433306

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00433306

Keywords

Navigation