1887

Abstract

DNA in the terminal region of the chromosome of was labelled by a procedure in which cells in sporulation inducing conditions were pulse-labelled with [H]thymidine and then treated with -hydroxyphenylazouracil, an inhibitor of DNA synthesis. The labelled DNA in isolated spores yielded a small number of restriction fragments. About 14 RI fragments with a total length of 80 kb were labelled in a 2·5 min pulse. A fragment of 4·0 kb had the highest specific radioactivity in terminally labelled DNA from several strains. One of these strains lacked the 120 kb prophage of SP that is normally integrated close to the terminus. Loss of the 120 kb prophage did not affect the point of termination which must therefore be regarded as a specific ‘stop’ sequence. Labelled terminus DNA has been used to identify lambda (Charon 4A) clones containing sequences derived from the terminal region. The total length of the restriction fragments present was 150 kb and adds another 90 kb to the 150 kb region mapped previously. Only one group of these sequences was present in a strain (CU1695) carrying a deletion spanning from SP to the right of the terminus and This suggests that the terminator sequence found in the wild-type can be deleted but presumably this strain has an alternative mechanism of termination.

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1984-09-01
2024-04-24
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References

  1. Adams R. T., Wake R. G. 1980; Highly specific labelling of the terminus of the Bacillus subtilis chromosome. Journal of Bacteriology 143:1036–1038
    [Google Scholar]
  2. Benton W. D., Davis R. W. 1977; Screening lambda gt. recombinant clones by hybridisation to single plaques in situ. Science 196:180–182
    [Google Scholar]
  3. Bouché J. P. 1982; Physical map of a 470 × 103 base pair region flanking the terminus of DNA replication in the Escherichia coli K12 genome. Journal of Molecular Biology 154:9–20
    [Google Scholar]
  4. Bouché J. P., Gelugne J. P., Louarn J., Louarn J. M., Kaiser K. 1982; Relationships between the physical and genetic maps of a 470 x 103 base pair region around the terminus of Escherichia coli K12. Journal of Molecular Biology 154:21–32
    [Google Scholar]
  5. Callister H., Wake R. G. 1974; Completed chromosomes in thymine requiring Bacillus subtilis spores. Journal of Bacteriology 120:579–582
    [Google Scholar]
  6. Davern C. 1979; Replication of the prokaryotic chromosome with emphasis on the bacterial chromosome replication in relation to the cell cycle. In Cell Biology 2 pp. 131–169 Prescott D. M., Goldstein L. Edited by New York: Academic Press;
    [Google Scholar]
  7. Deshpande K. L., Kan J. F. 1980; Glutamate synthase from Bacillus subtilis: in vitro reconstitution of an active amido-transferase. Biochemical and Biophysical Research Communications 93:308–314
    [Google Scholar]
  8. Dewet J. R., Daniels D. L., Schroeder J. L., Williams B. G., Denniston-Thompson K., Moore D. D., Blattner F. R. 1980; Restriction maps for twenty-one Charon vector phages. Journal of Virology 33:401–410
    [Google Scholar]
  9. Dretzen G., Bellard M., Sassone-CORSI P., Chambon P. 1981; A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Analytical Biochemistry 112:295–298
    [Google Scholar]
  10. Ferrari E., Henner D. J., Hoch J. A. 1981; Isolation of Bacillus subtilis genes from a Charon 4A library. Journal of Bacteriology 146:430–432
    [Google Scholar]
  11. Fisher R. S., Kane J. F. 1981; Regulation of glutamate synthase in Bacillus subtilis. Proceedings of the CETUS Conference on Molecular Cloning and Gene Regulation in Bacilli. Stanford University;
    [Google Scholar]
  12. Germino J., Bastia D. 1981; Termination of DNA replication in vitro at a sequence specific replication terminus. Cell 23:681–687
    [Google Scholar]
  13. Harford N. 1975; Bidirectional chromosome replication in Bacillus subtilis. Journal of Bacteriology 121:835–847
    [Google Scholar]
  14. Henner D. J., Hoch J. A. 1980; The Bacillus subtilis chromosome. Microbiological Reviews 44:57–82
    [Google Scholar]
  15. Jeffreys A. L., Flavell R. A. 1977; A physical map of the DNA regions flanking the rabbit β-globin genes. Cell 12:429–439
    [Google Scholar]
  16. Kuempel P. L., Duerr S. A., Seeley N. R. 1977; Terminus region of the chromosome in Escherichia coli inhibits replication forks. Proceedings of the National Academy of Sciences of the United States of America 74:3927–3931
    [Google Scholar]
  17. Laskey R. A. 1980; The use of intensifying screens or organic scintillators for visualising radioactive molecules resolved by gel electrophoresis. Methods in Enzymology 65:363–370
    [Google Scholar]
  18. Louarn J., Patte J., Louarn J. M. 1977; Evidence for a fixed termination site of chromosome replication in Escherichia coli K12. Journal of Molecular Biology 155:295–314
    [Google Scholar]
  19. O’SULLIVAN M. A., Anagnostopoulos C. 1982; Replication terminus of the Bacillus subtilis chromosome. Journal of Bacteriology 151:135–143
    [Google Scholar]
  20. Reysset G. 1981; New class of Bacillus subtilis glutamine requiring mutants. Journal of Bacteriology 148:653–658
    [Google Scholar]
  21. Rosenthal R., Toye P. A., Korman R. Z., Zahler S. A. 1979; The prophage of SP/Jc2 CitK,. A defective specialised transducing phage of B. subtilis. Genetics 92:721–739
    [Google Scholar]
  22. Sargent M. G. 1980a; Chromosome replication in sporulating cells of Bacillus subtilis. Journal of Bacteriology 142:491–498
    [Google Scholar]
  23. Sargent M. G. 1980b; Specific labelling of the Bacillus subtilis terminus. Journal of Bacteriology 143:1033–1035
    [Google Scholar]
  24. Sargent M. G. 1980c; A procedure for isolating high quality DNA from spores of Bacillus subtilis. Journal of General Microbiology 116:511–514
    [Google Scholar]
  25. Sargent M. G., Bennett M. F. 1982; Attachment of the chromosomal terminus of Bacillus subtilis to a fast sedimenting particle. Journal of Bacteriology 150:623–632
    [Google Scholar]
  26. Sargent M. G., Monteiro M. J. 1982; Characterization of the chromosomal terminus of B. subtilis and its attachment to the cell membrane. In Molecular Cloning and Gene Regulation in Bacilli pp. 181–195 Ganesan A. T., Chang S., Hoch J. A. Edited by New York: Academic Press;
    [Google Scholar]
  27. Sargent M. G., Bennett M. S., Burdett I. D. J. 1983; Identification of specific restriction fragments associated with a membrane sub-particle from Bacillus subtilis. Journal of Bacteriology 154:1389–1396
    [Google Scholar]
  28. Southern E. 1979; Gel electrophoresis of restriction fragments. Methods in Enzymology 68:152–176
    [Google Scholar]
  29. Warner F. G., Kotos G. A., Romano M. P., Hemphill H. E. 1977; Characterisation of SPβ, a temperate phage of Bacillus subtilis Ml68. Canadian Journal of Microbiology 129:556–558
    [Google Scholar]
  30. Weiss A. S. 1983; Restriction map of DNA spanning the replication terminus of the Bacillus subtilis chromosome. Journal of Molecular Biology 171:119–137
    [Google Scholar]
  31. Weiss A. S., Hariharan I. K., Wake R. G. 1981; Analysis of the terminus region of the Bacillus subtilis chromosome. Nature; London: 293673–675
    [Google Scholar]
  32. Weiss A. S., Hariharan I. K., Adams R. T., Wake R. G. 1982; Characterisation of the terminus region of the B. subtilis chromosome. In Molecular Cloning and Gene Regulation in Bacilli pp. 197–210 Ganesan A. T., Chang S., Hoch J. A. Edited by New York: Academic Press;
    [Google Scholar]
  33. Zahler S. A. 1982; Specialised transduction in Bacillus subtilis. . In The Molecular Biology of the Bacilli pp. 269–305 Dubnau D. A. Edited by New York: Academic Press;
    [Google Scholar]
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