1887

Abstract

SUMMARY

To study the effect of gene dosage on gene expression, λp5I857293, a replication defective lambda phage carrying part of the operon (containing the promoter, operator and gene) in the b2 region was studied in strain JC6256 where the operon is deleted and at a temperature where the λ repressor is inactive. In measuring the synthesis of β-galactosidase, it was possible to separate the effects of the promoter from those of the phage promoter. When the synthesis of β-galactosidase was initiated from the inserted promoter in JC6256(λ) in the presence of additional cyclic AMP, the rate and level of β-galactosidase synthesis were directly proportional to the multiplicity of infection (gene dosage). Furthermore, β-galactosidase synthesis was initiated about 5 min after infection, just as with isopropyl-β--thiogalactoside (IPTG) induction.

When the synthesis of β-galactosidase was initiated from the phage promoter in JC6256 in the absence of additional cyclic AMP, the rate and level of β-galactosidase synthesis were again linearly proportional to gene dosage. On the other hand, initiation of β-galactosidase synthesis was delayed until 10 to 20 min after infection. These results suggest that: (i) in the absence of negative controlling factors, the extent of gene expression is proportional to gene dosage; (ii) varying the gene dosage can be used to regulate gene expression.

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1982-02-01
2022-12-08
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References

  1. Blattner F. R., Williams B. G., Blechl A. E., Denniston-Thompson K., Faber H. E., Furlong L., Grunwald D. J., Kiefer D. O., Moore D. D., Schumn J. W., Scheldon E. L., Smithies O. 1977; Charon phages: safer derivatives of bacteriophage lambda for DNA cloning. Science 196:161–169
    [Google Scholar]
  2. Brunschede H., Bremer H. 1976; Synthesis of phage- and host-specific RNA in Escherichia coli infected with a fragment of bacteriophage T5. Virology 15:355–367
    [Google Scholar]
  3. Campbell A. 1961; Sensitive mutants of bacteriophage. X. Virology 14:22–32
    [Google Scholar]
  4. Chandler M. G., Pritchard R. H. 1975; The effect of gene concentration and relative gene dosage on gene output in Escherichia coli. Molecular and General Genetics 138:127–141
    [Google Scholar]
  5. Cohen S. N., Chang A. C. Y. 1970; Genetic expression in bacteriophage. III. Inhibition of Escherichia coli nucleic acid and protein synthesis during development. Journal of Molecular Biology 49:557–575
    [Google Scholar]
  6. Dove W. F. 1966; Action of the lambda chromosome. I. Control of functions late in bacteriophage development. Journalof Molecular Biology 19:187–201
    [Google Scholar]
  7. Echols H. 1974; Some unsolved general problems of phage X development. Biochimie 56:1491–1496
    [Google Scholar]
  8. Echols H., Green L., Oppenheim A. B., Oppenheim A., Honigman A. 1973; Role of the cro gene in bacteriophage X development. Journal of Molecular Biology 80:203–216
    [Google Scholar]
  9. Edstrom J. E., Lambert B. 1975; Gene and information diversity in eukaryotes. Progress in Biophysics and Molecular Biology 30:57–82
    [Google Scholar]
  10. Eicher E. M., Coleman D. L. 1977; Influence of gene duplication and X-inactivation on mouse mitochondrial malic enzyme activity and electrophoretic patterns. Genetics 85:647–658
    [Google Scholar]
  11. Eisen H., Ptashne M. 1971; Regulation of repressor synthesis. In The Bacteriophage Lambda pp 239–245 Edited by Hershey A. D. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  12. Eisen H., Brachet P., Pereira Da Silva L., Jacob F. 1970; Regulation of repressor expression in X. Proceedings of the National Academy of Sciences of the United States of America 66:855–862
    [Google Scholar]
  13. Epp G., Pearson M. L. 1976; Association of bacteriophage lambda N gene protein with E. coli RNA polymerase. In RNA Polymerase pp 667–691 Edited by Chamberlin M., Losick R. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  14. Epstein C. J., Tucker G., Travis B., Gropp A. 1977; Gene dosage for isocitrate dehydrogenase in mouse embryos trisomic for chromosome 1. Nature, London 267:615–616
    [Google Scholar]
  15. Hayes S. 1979; Initiation of coliphage lambda replication, lit, oop RNA synthesis, and effect of gene dosage on transcription from promoters PL, PR and PR′. Virology 97:415–438
    [Google Scholar]
  16. Horiuchi T., Ohshima Y. 1966; Inhibition of repressor formation in the lactose system of Escherichia coli by inhibition of protein synthesis. Journal of Molecular Biology 20:517–526
    [Google Scholar]
  17. Hourcade D., Dressler D., Wolfson J. 1973; The nucleolus and the rolling circle. Cold Spring Harbor Symposia on Quantitative Biology 38:537–550
    [Google Scholar]
  18. Ippen K., Shapiro J. A., Beckwith J. R. 1971; Transposition of the lac region to the gal region of the Escherichia coli chromosome: isolation of lac transducing bacteriophages. Journal of Bacteriology 108:5–9
    [Google Scholar]
  19. Kaiser A. D. 1957; Mutations in a temperate bacteriophage affecting its ability to lysogenize Escherichia coli. Virology 3:42–61
    [Google Scholar]
  20. Kaiser A. D., Jacob F. 1957; Recombination between related temperate bacteriophages and the genetic control of immunity and prophage localization. Virology 4:509–521
    [Google Scholar]
  21. Luk K. C., Mark K. K. 1979; The separation of phage promoter from bacterial lac promoter for β-galactosidase expression in transducing phage Ap/ac5. Biochemical and Biophysical Research Communications 89:64–70
    [Google Scholar]
  22. Luk K. C., Mark K. K. 1980; The phage promoter responsible for the expression of the inserted β-galactosidase gene in bacteriophage λplac 5. Molecular and General Genetics 178:555–560
    [Google Scholar]
  23. Malamy M. H., Fiandt M., Szybalski W. 1972; Electron microscopy of polar insertions in the lac operon of Escherichia coli. Molecular and General Genetics 119:207–222
    [Google Scholar]
  24. Mark K. K. 1970; The relationship between the synthesis of DNA and the synthesis of phage lysozyme in Escherichia coli infected by bacteriophage T4. Virology 42:20–27
    [Google Scholar]
  25. Mercereau-Puualon O., Kourilsky P. 1976; Escape synthesis of /J-galactosidase under the control of bacteriophage lambda. Journal of Molecular Biology 108:733–751
    [Google Scholar]
  26. Mow A N. R., Katz E., Adsourian P. L., Hirota Y., Inouye M. 1978; Gene dosage effects of the structural gene for a lipoprotein of the Escherichia coli outer membrane. Journal of Bacteriology 133:81–84
    [Google Scholar]
  27. Nijkamp H. J. J., Bovre K., Szybalski W. 1970; Controls of rightward transcription in coliphage l. Journal of Molecular Biology 54:599–604
    [Google Scholar]
  28. Ogawa T., Tomizawa U. 1968; Replication of bacteriophage DNA. I. Replication of DNA of lambda phage defective in early function. Journal of Molecular Biology 38:217–225
    [Google Scholar]
  29. Pardee A. B., Jacob F., Monod J. 1959; The genetic control and cytoplasmic expression of inducibility in the synthesis of β-galactosidase in E. coli. Journal of Molecular Biology 1:165–178
    [Google Scholar]
  30. Pero J. 1970; Location of the phage A gene responsible for turning off A-exonuclease synthesis. Virology 40:65–71
    [Google Scholar]
  31. Putnam S. L., Koch A. L. 1975; Complication in the simplest cellular enzyme assay: lysis of Escherichia coli for the assay of β-galactosidase. Analytical Biochemistry 63:350–360
    [Google Scholar]
  32. Ray P. N., Pearson M. L. 1976; Synthesis of morphogenetic proteins by mutants of bacteriophage lambda carrying tandem genetic duplications. Virology 73:381–388
    [Google Scholar]
  33. Reichardt L. 1975a; Control of bacteriophage lambda repressor synthesis after phage infection: the role of N. ell, cIII and cro products. Journal of Molecular Biology 93:267–288
    [Google Scholar]
  34. Reichardt L. 1975b; Control of bacteriophage lambda repressor synthesis: regulation of the maintenance pathway by the cro and cl products. Journal of Molecular Biology 93:289–309
    [Google Scholar]
  35. Reichert U., Winter M. 1975; Gene dosage effects in polyploid strains of Saccharomyces cerevisiae containing gua-1 wild-type and mutant alleles. Journal of Bacteriology 124:1041–1045
    [Google Scholar]
  36. Shapiro J., Machattie L., Eron L., Ihler G., Ippen K., Beckwith J. 1969; Isolation of pure lac operon DNA. Nature, London 224:768–774
    [Google Scholar]
  37. Szybalski W. 1972; Transcription and replication in E. coli bacteriophage lambda. In Uptake of Informative Molecules by Living Cells pp 59–82 Edited by Ledoux L. Amsterdam: North-Holland;
    [Google Scholar]
  38. Szybalski W. 1976; Genetic and molecular map of Escherichia coli bacteriophage lambda (A). In Handbook of Biochemistry and Molecular Biology 3rd edn pp 677–685 Edited by Fasman G. D. Nucleic Acids vol 2: Cleveland: CRC Press;
    [Google Scholar]
  39. Takeda Y. 1975; Regulation of early protein synthesis in bacteriophage A. Virology 66:629–631
    [Google Scholar]
  40. Takeda Y., Matsubara K., Ogata K. 1975; Regulation of early gene expression in bacteriophage lambda: effect of tof mutation on strand-specific transcription. Virology 65:374–384
    [Google Scholar]
  41. Tartof K. D. 1975; Redundant genes. Annual Review of Genetics 9:355–385
    [Google Scholar]
  42. Tsui L. C., Mark K. K. 1976; The depression of endolysin synthesis in bacteria infected with high multiplicities of phage A. Molecular and General Genetics 143:269–278
    [Google Scholar]
  43. Wu A. M., Ghosh S., Willard M., Davison J., Echols H. 1971; Negative regulation by lambda: repression of lambda RNA synthesis in vitro and host enzyme synthesis in vivo. In The Bacteriophage Lambda pp 589–598 Edited by Hershey A. D. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
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