1887

Abstract

SUMMARY: Fusions of the structural genes to the proximal promoters and of the operon have been isolated using a λ (lac, Mu') phage. This phage was inserted into a heat-inducible Mu prophage which had itself been inserted into the gene. Selection was made for heat-resistant derivatives of this lysogen which maximally expressed the genes when an inducer of the operon was present; these were shown to have the genes fused to the operon - in some cases with a concomitant deletion of the gene, whilst in others was retained. The level of β-galactosidase activity in these strains was induced by growth in the presence of deoxyadenosine or cytidine, both of which lead to induction of the enzymes - deoxyadenosine through the system and cytidine through the system. The level of induction of β-galactosidase was of the same order as the level of induction of deoxyriboaldolase, the gene product, in those strains retaining an intact gene. Plaque-forming λ phages carrying the fusions were isolated by inducing the lysogens with mitomycin C.

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1979-06-01
2024-04-25
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References

  1. Ahmad S. I., Pritchard R. H. 1971; A regulatory mutant affecting the synthesis of enzymes involved in the catabolism of nucleosides in Escherichia coli . Molecular and General Genetics 111:77–83
    [Google Scholar]
  2. Ahmad S. I., Pritchard R. H. 1973; An operator constitutive mutant affecting the synthesis of two enzymes involved in the catabolism of nucleosides in Escherichia coli . Molecular and General Genetics 124:321–328
    [Google Scholar]
  3. Albrechtsen H., Hammer-Jespersen K., Munch-Petersen A., Fill N. 1976; Multiple regulation of nucleoside catabolising enzymes: effects of a polar dra mutation on the deo enzymes. Molecular and General Genetics 146:139–145
    [Google Scholar]
  4. Baumanis G. E., Smirnov Y. V., Sukhodoletz V. V. 1974; Production and study of polar mutants for nucleoside catabolism linked genes in Escherichia coli . Genetika 10:81–88
    [Google Scholar]
  5. Beckwith J. 1970; Lac: the genetic system. In The Lactose Operon pp 5–26 Edited by Beckwith J. R., Zipser D. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  6. Breitman T. R., Bradford R. M. 1968; Inability of low-thymine requiring mutants of Escherichia coli lacking phosphodeoxyribomutase to be induced for deoxythymidine phosphorylase and deoxyriboaldolase. Journal of Bacteriology 95:2434–2435
    [Google Scholar]
  7. Burton K. 1956; A study of the conditions and mechanism of the diphenylamine reaction for the calorimetric estimation of deoxyribonucleic acid. Biochemical Journal 62:315–323
    [Google Scholar]
  8. Buxton R. S. 1975; Genetic analysis of thymidine-resistant and low-thymine-requiring mutants of Escherichia coli K-12 induced by bacteriophage Mu-1. Journal of Bacteriology 121:475–484
    [Google Scholar]
  9. Buxton R. S., Albrechtsen H., Hammer-Jespersen K. 1977; Overlapping transcriptional units in the deo operon of Escherichia coli K-12. Evidence from phage Mu-1 insertion mutants. Journal of Molecular Biology 114:287–300
    [Google Scholar]
  10. Buxton R. S., Hammer-Jespersen K., Hansen T. D. 1978; Insertion of bacteriophage lambda into the deo operon of Escherichia coli K-12 and the isolation of plaque-forming λdeo+ transducing phages. Journal of Bacteriology 136:668–681
    [Google Scholar]
  11. Casadaban M. J. 1976; Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. Journal of Molecular Biology 104:541–555
    [Google Scholar]
  12. Hammer-Jespersen K., Munch-Petersen A. 1973; Mutants of Escherichia coli unable to metabolise cytidine: isolation and characterisation. Molecular and General Genetics 126:177–186
    [Google Scholar]
  13. Hammer-Jespersen K., Munch-Petersen A. 1975; Multiple regulation of nucleoside catabolizing enzymes: regulation of the deo operon by the cytR and deoR gene products. Molecular and General Genetics 137:327–335
    [Google Scholar]
  14. Hammer-Jespersen K., Nygaard P. 1976; Multiple regulation of nucleoside catabolizing enzymes in Escherichia coli: effects of 3:5ʹ cyclic AMP and CRP protein. Molecular and General Genetics 148:49–55
    [Google Scholar]
  15. Hopkins J. 1974; A new class of promoter mutations in the lactose operon of Escherichia coli . Journal of Molecular Biology 87:715–724
    [Google Scholar]
  16. Jorgensen P., Collins J., Valentin-Hansen P. 1977; On the structure of the deo operon of Escherichia coli . Molecular and General Genetics 155:93–102
    [Google Scholar]
  17. Lindahl G., Sironi G., Bialy H., Calendar R. 1970; Bacteriophage lambda: abortive infection of bacteria lysogenic for phage P2. Proceedings of the National Academy of Sciences of the United States of America 66:587–594
    [Google Scholar]
  18. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  19. Miller J. H. 1972 Experiments in Molecular Genetics p 403 New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Munch-Petersen A., Nygaard P., Hammer-Jespersen K., Fiil N. 1972; Mutants constitutive for nucleoside-catabolizing enzymes in Escherichia coli K12. European Journal of Biochemistry 27:208–215
    [Google Scholar]
  21. Rosner J. L. 1972; Formation, induction and curing of bacteriophage P1 lysogens. Virology 48:679–689
    [Google Scholar]
  22. Svenningsen B. A. 1975; Regulated in vitro synthesis of the enzymes of the deo operon of Escherichia coli. Properties of the DNA directed system. Molecular and General Genetics 137:289–304
    [Google Scholar]
  23. Valentin-Hansen P., Svenningsen B. A., Munch-Petersen A., Hammer-Jespersen K. 1978; Regulation of the deo operon in Escherichia coli. The double negative control of the deo operon by the cytR and deoR repressors in a DNA directed in vitro system. Molecular and General Genetics 159:191–202
    [Google Scholar]
  24. Zeldis J. B., Bukhari A. I., Zipser D. 1973; Orientation of prophage Mu. Virology 55:289–294
    [Google Scholar]
  25. Zissler J., Signer E., Schaefer F. 1971; The role of recombination in growth of bacteriophage lambda. I. The gamma gene. In The Bacteriophage Lambda pp 455–468 Edited by Hershey A. D. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
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