1887

Abstract

The gene of , which encodes the transcriptional activator of gene expression, was cloned into a number of plasmid vectors to obtain high-level synthesis of product (NifA). When over-produced, NifA was very insoluble and it precipitated with the cell debris after cell lysis. Localization of -galactosidase activity from a translational fusion confirmed the insoluble nature of NifA. Analysis of two translational fusions in which the last six C-terminal amino acids of NifA were deleted suggests that these residues are required for activity.

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/content/journal/micro/10.1099/00221287-134-2-425
1988-02-01
2021-05-15
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References

  1. Austin S., Henderson N., Dixon R. 1987; Requirements for transcriptional activation in vitro of the nitrogen regulated glnA and nifLA promoters from Klebsiellapneumoniae: dependence on activator concentration. Molecular Microbiology 1:92–100
    [Google Scholar]
  2. Beynon J., Cannon , Buchanan-Wollaston V., Cannon F. 1983; The nif promoters of Klebsiella pneumoniae have a characteristic primary structure. Cell 44:665–671
    [Google Scholar]
  3. Brooks S. J., Collins J. J., Brill W. J. 1984; Repression of nitrogen fixation in Klebsiella pneumoniae at high temperature. Journal of Bacteriology 157:460–464
    [Google Scholar]
  4. Buchanan-Wollaston V., Cannon M. C., Beynon J. L., Cannon F. C. 1981; Role of the nifA gene product in the regulation of nif expression in Klebsiella pneumoniae. Nature; London: 294776–778
    [Google Scholar]
  5. Buck M., Cannon W. 1987; Frameshifts close to the Klebsiella pneumoniaenifH promoter prevent multicopy inhibition by hybrid nifH plasmids. Molecular and General Genetics 207:492–498
    [Google Scholar]
  6. Buck M., Miller S., Drummond M., Dixon R. 1986; Upstream activator sequences are present in the promoters of nitrogen fixation genes. Nature; London: 320374–378
    [Google Scholar]
  7. Buck M., Cannon W., Woodcock J. 1987; Transcriptional activation of the Klebsiella pneumoniae nitrogenase promoter may involve DNA loop formation. Molecular Microbiology 1:243–249
    [Google Scholar]
  8. Buikema W. J., Szeto W. W., Lemley P. V., Orme-Johnson W. H., Ausubel F. M. 1985; Nitrogen fixation specific regulatory genes of Klebsiella pneumoniae and Rhizobium meliloti share homology with the general nitrogen regulatory gene ntrC of K.pneumoniae. Nucleic Acids Research 12:4539–4555
    [Google Scholar]
  9. Casadaban M. J., Cohen S. N. 1980; Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. Journal of Molecular Biology 138:179–207
    [Google Scholar]
  10. Collins J. 1979; Cell free synthesis of proteins coding for mobilisation functions of ColEl and transposition functions of Tn3. Gene 6:29–42
    [Google Scholar]
  11. Collins J. J., Roberts G. P., Brill W. J. 1986; Posttranscriptional control of Klebsiella pneumoniaenif mRNA stability by the nifL product. Journal of Bacteriology 168:173–178
    [Google Scholar]
  12. Drummond M., Wootton J. 1987; Sequence of nifL from Klebsiellapneumoniae: mode of action and relationship to two families of regulatory proteins. Molecular Microbiology 1:37–44
    [Google Scholar]
  13. Drummond M., Whitty P., Wootton J. 1986; Sequence and domain relationships of ntrC and nifA from Klebsiellapneumoniae: homologies to other regulatory proteins. EMBO Journal 5:441–447
    [Google Scholar]
  14. Espin G., Alvarez-Morales A., Cannon F., Dixon R., Merrick M. 1982; Cloning of the gin A, ntrB and ntrC genes of Klebsiellapneumoniae and studies on their role in regulation of the nitrogen fixation (nif) gene cluster. Molecular and General Genetics 186:518–524
    [Google Scholar]
  15. Hawkes T., Merrick M., Dixon R. 1985; Interaction of purified NtrC protein with nitrogen regulated promoters from Klebsiella pneumoniae. Molecular and General Genetics 201:492–498
    [Google Scholar]
  16. Hill S., Kennedy C., Kavanagh E., Goldberg R. B., Hanau R. 1981; Nitrogen fixation gene (nifL) involved in oxygen regulation of nitrogenase synthesis in Klebsiella pneumoniae. Nature; London: 290424–426
    [Google Scholar]
  17. Hirschman J., Wong P. K., Sei K., Keener J., Kustu S. 1985; Products of nitrogen regulatory genes ntrA and ntrC of enteric bacteria activate glnA transcription in vitro: evidence that the ntrA product is a sigma factor. Proceedings of the National Academy of Sciences of the United States of America 82:7525–7529
    [Google Scholar]
  18. Hunt T. P., Magasanik B. 1985; Transcription of glnA by purified Escherichia coli components: core RNA polymerase and the products of glnF, glnG and glnL. Proceedings of the National Academy of Sciences of the United States of America 82:8453–8457
    [Google Scholar]
  19. Kröger M., Hobom G. 1982; Structural analysis of insertion sequence IS5. Nature; London: 297:159–162
    [Google Scholar]
  20. Merrick M. 1983; Nitrogen control of the nif regulon in Klebsiella pneumoniae : involvement of the ntrA gene and analogies between ntrC and nifA. EMBO Journal 2:39–44
    [Google Scholar]
  21. Merrick M., Hill S., Hennecke H., Hahn M., Dixon R., Kennedy C. 1982; Repressor properties of the nifL gene product in Klebsiella pneumoniae. Molecular and General Genetics 185:75–81
    [Google Scholar]
  22. Minton N. P. 1984; Improved plasmid vectors for the isolation of translational lac gene fusions. Gene 31:269–273
    [Google Scholar]
  23. Ow D. W., Ausubel F. M. 1983; The nifA gene which regulates the Klebsiellapneumoniaenif gene cluster can substitute for the nitrogen regulatory gene glnG (ntrC). Nature; London: 301307–313
    [Google Scholar]
  24. Putnam S. L., Koch A. L. 1975; Complications in the simplest cellular enzyme assay: lysis of Escherichia coli for the assay of β-galactosidase. Analytical Biochemistry 63:350–360
    [Google Scholar]
  25. Remaut E., Stanssens P., Fiers W. 1981; Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. Gene 15:81–93
    [Google Scholar]
  26. Ronson C. W., Nixon B. T., Ausubel F. M. 1987; Conserved domains in bacterial regulatory proteins that respond to environmental stimuli. Cell 49:579–581
    [Google Scholar]
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