Skip to main content
Log in

Polarity of mutations induced by insertion of transposons Tn5, Tn7 and Tn10 into the nif gene cluster of Klebsiella pneumoniae

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

Summary

Three new genes nifM, nifI and nifN have been mapped in the nif gene cluster of Klebsiella pneumoniae and a fourth gene nifJ has been confirmed as being a separate cistron. Polar nif mutations were obtained by transposition of Tn7 to plasmid pRD1, and of Tn5 and Tn10 to plasmid pMF100, a derivative of pRD1. Complementation analysis of the nif::Tn mutants led to the identification of at least six transcriptional units: nifB; nifA; nifJ; nifH, nifD and nifK; nifE and nifI; nifN, nifM and nifF. Biochemical and genetic evidence suggest that the three genes nifH, nifD and nifK, which are probably the structural genes for nitrogenase, belong to the same operon and are transcribed from nifH to nifK. A polypeptide with a molecular weight of approximately 120,000 is presumed to be the nifJ product.

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

  • Atkins, J.F., Loper, J.C.: Transcription initiation in the histidine operon of Salmonella typhimurium. Proc. nat. Acad. Sci. (Wash.) 65, 925–932 (1970)

    Google Scholar 

  • Bachhuber, M., Brill, W.J., Howe, M.M.: Use of bacteriophage Mu to isolate deletions in the his-nif region of Klebsiella pneumoniae. J. Bact. 128, 749–753 (1976)

    Google Scholar 

  • Barth, P.T., Datta, N., Hedges, R.W., Grinter, N.J.: Transposition of a deoxyribonucleic acid sequence encoding trimethoprim and streptomycin resistances from R483 to other replicons. J. Bact. 125, 800–810 (1976)

    Google Scholar 

  • Barth, P.T., Grinter, N.J.: Map of plasmid RP4 derived by insertion of transposon TnC. J. molec. Biol. 113, 455–474 (1977)

    Google Scholar 

  • Berg, D.: Insertion and excision of the transposable kanamycin resistance determinant Tn5. In: DNA insertion elements, plasmids and episomes (A.T. Bukhari, J.A. Shapiro and S. Adhya, eds.). New York: Cold Spring Harbor Laboratory 1977

    Google Scholar 

  • Berg, D., Davies, J., Allet, B., Rochaix, J.D.: Transposition of R factor genes to bacteriophage λ. Proc. nat. Acad. Sci. (Wash.) 72, 3628–3632 (1975)

    Google Scholar 

  • Besemer, J., Herpers, M.: Suppression of polarity of insertion mutations within the gal operon of E. coli. Molec. gen. Genet. 151, 295–304 (1977)

    Google Scholar 

  • Botstein, D., Kleckner, N.: Translocation and illegitimate recombination by the tetracycline resistance element Tn10. In: DNA insertion elements, plasmids and episomes (A.T. Bukhari, J.A. Shapiro, and S. Adhya, eds.). New York: Cold Spring Harbor Laboratory 1977

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Dixon, R.A., Postgate, J.R.: Genetic transfer of nitrogen fixation from Klebsiella pneumoniae to Escherichia coli. Nature (Lond.) 237, 102–103 (1972)

    Google Scholar 

  • Eady, R.R., Issack, R., Kennedy, C., Postagate, J.R., Ratcliffe, H.D.: Nitrogenase synthesis in Klebsiella pneumoniae: Comparison of regulation by NH4 + and oxygen. J. gen. Microbiol. 104, 277–285 (1978)

    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, 655–657 (1972)

    Google Scholar 

  • 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. Molec. gen. Genet. (in press)

  • Foster, T.J.: Insertion of the tetracycline resistance translation unit Tn10 in the lac operon of Escherichia coli K12. Molec. gen. Genet. 154, 305–309 (1977)

    Google Scholar 

  • Foster, T.J., Howe, T.G.B., Richmond, K.M.V.: Translocation of the tetracycline resistance determinant from R100-1 to the E. coli K12 chromosome. J. Bact. 124, 1153–1158 (1975)

    Google Scholar 

  • Foster, T.J., Willetts, N.S.: Characterisation of transfer-deficient mutants of the R100-1 Tcs plasmid pDU202, caused by insertion of Tn10. Molec. gen. Genet. 156, 107–114 (1977)

    Google Scholar 

  • Goldberg, R.V., Bender, R.A., Streicher, S.L.: Direct selection for PI-sensitive mutants of enteric bacteria. J. Bact. 118, 810–814 (1974)

    Google Scholar 

  • Jordan, E., Saedler, H., Starlinger, P.: Strong polar mutations in the transferase gene of the galactose operon in E. coli. Molec. gen. Genet. 100, 296 (1967)

    Google Scholar 

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

    Google Scholar 

  • Kennedy, C., Eady, R.R., Kondorosi, E., Klavans-Redosh, D.: The molybdenum-iron protein of Klebsiella pneumoniae: evidence for non-identical sub-units from peptide mapping. Biochem. J. 155, 383–389 (1976)

    Google Scholar 

  • Kleckner, N.: Translocatable elements in prokaryotes. Cell 11, 11–23 (1977)

    Google Scholar 

  • Kleckner, N., Chan, R.K., Tye, B.-K., Botstein, D.: Mutagenesis by insertion of a drug-resistance element carrying an inverted repetition. J. molec Biol. 97, 561–575 (1975)

    Google Scholar 

  • Kleckner, N., Roth, J., Botstein, D.: Genetic engineering in vivo using translocatable drug-resistance elements. J. molec. Biol. 116, 125–159 (1977)

    Google Scholar 

  • St. John, R.T., Johnston, M.H., 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. Bact. 121, 759–765 (1975)

    Google Scholar 

  • Streicher, S., Guerney, E.G., Valentine, R.C.: The nitrogen fixation genes. Nature (Lond.) 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

Merrick, M., Filser, M., Kennedy, C. et al. Polarity of mutations induced by insertion of transposons Tn5, Tn7 and Tn10 into the nif gene cluster of Klebsiella pneumoniae . Molec. Gen. Genet. 165, 103–111 (1978). https://doi.org/10.1007/BF00270382

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00270382

Keywords

Navigation