1887

Abstract

Summary: In several strains of extensive breakdown of chromosomal DNA may be potentiated by osmotic lysisof protoplasts. At its most severe, in strains originating from Farmer& Rothman’s thymine auxotroph, the rate of DNA breakdown was greater than 50% per hour at 40 °C. The rate of DNA breakdown in most other strains tested was approximately 5% per hour except for SP β-strains, in which the rate of DNA breakdown was only 0.3 %. DNA degradation was attributed to relaxation ofcontrol of a nuclease specified by the prophage of SP or a related phage. The most potent nuclease in lysates was an ATP-activated protein of 280000. Derivatives of Farmer and Rothman’s strain containing integrated plasmids had the highest rate of DNA degradation. Although the chromosome was completely destroyed, covalently closed circular plasmids were generated from the integrated sequence. These showed massive deletions of the part of the integrated plasmid but the vector sequence remained intact. The nucleolytic activity therefore appears to recognize specific sequences in DNA. We suggest that activation of SP genes during development of competence may be a cause of deletion ofcloned genes in the early stages of establishment of cloned sequences.

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1985-10-01
2021-08-02
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References

  1. Birnboim H.C. 1966; Cellular site in B. subtilis of a nuclease which preferentially degrades single stranded nucleic acids. Journal of Bacteriology 91:1005–1011
    [Google Scholar]
  2. Bonamy C., Szulmajster J. 1982; Cloning and expression of B. subtilis spore genes. Molecular and General Genetics 188:202–210
    [Google Scholar]
  3. Buxton R.S. 1980; Selection of Bacillus subtilis 168 mutants with deletions of the PBSX prophage. Journal of General Virology 46:427–437
    [Google Scholar]
  4. Callister H., Wake R.G. 1974; Completed chromosomes in thymine-requiring Bacillus subtilis spores. Journal of Bacteriology 120:579–582
    [Google Scholar]
  5. Dedonder R.A., Lepesant J.A., Lepesant.Kejzlarova J., Billault A., Steinmetz M., Kunst F. 1977; Construction of a kit of reference strains for rapid genetic mapping in B. subtilis 168. Applied and Environmental Microbiology 33:989–993
    [Google Scholar]
  6. Doly J., Anagnostopoulos C. 1976; Isolation,subunit structure and properties of the ATP dependent DNAase of Bacillus subtilis. European Journal of Biochemistry 71:309–316
    [Google Scholar]
  7. Farmer J.L., Rothman F. 1965; Transformable thymine-requiring mutant of Bacillus subtilis. Journal of Bacteriology 89:262–263
    [Google Scholar]
  8. Ferrari F.A., Nguyen A., Lang D., Hoch J.A. 1983; Construction and properties of an integrable plasmid for B. subtilis. Journal of Bacteriology 154:1513–1515
    [Google Scholar]
  9. Fugii M., Sakaguchi K. 1980; A site-specific recE4 independent intramolecular recombination between Bacillus subtilis and Staphylococcus aureus DNA in a hybrid plasmid. Gene 12:95–102
    [Google Scholar]
  10. Ganesan A.T. 1979; Genetic recombination during transformation in Bacillus subtilis: appearance of a deoxyribonucleic acid methylase. Journal of Bacteriaology 139:270–279
    [Google Scholar]
  11. Haas M., Yoshikawa H. 1969; Defective bacteriophage PBSH in Bacillus subtilis. I. Induction, purification and physical properties of the bacteriophage and its DNA. Journal of Virology 3:233–247
    [Google Scholar]
  12. Hanahan D. 1983; Studies on transformation of Escherichia coli with plasmids. Journal of Molecular Biology 166:557–580
    [Google Scholar]
  13. Ish-Horowicz D., Burke J.F. 1981; Rapid efficient cosmid cloning. Nucleic Acids Research 9:2989–2998
    [Google Scholar]
  14. Kaiser K., Murray N E. 1979; Physical characterisation of the ‘Rac’ prophage in E. coli K12. Molecular and General Genetics 175:159–174
    [Google Scholar]
  15. Kerr I.M., Pratt E.A., Lehman I.R. 1965; Exonucleolytic degradation of high molecular weight DNA and RNA to nucleoside 3’-phosphates by a nuclease from Bacillus subtilis. Biochemical and Biophysical Research Communications 20:154–162
    [Google Scholar]
  16. Linn S. 1981; Deoxyribonucleases. Survey and Perspectives. In The Enzymes vol XIV121–135 Edited by Boyer P. D. London New York: Academic Press;
    [Google Scholar]
  17. Love E., D’ambrosio J., Brown N.C., Dubnau D. 1976; Mapping of the gene specifying DNA polymerase III of Bacillus subtilis. Molecular and General Genetics 144:313–321
    [Google Scholar]
  18. Mccarthy C., Nester E.W. 1969; Heat activated endonuclease in Bacillus subtilis. Journal of Bacteriology 87:1426–1430
    [Google Scholar]
  19. Ohi S., Sueoka N. 1973; Adenosine triphosphate dependent DNAase in Bacillus subtilis. I. Purification and some properties. Journal of Biological Chemistry 248:7336–7344
    [Google Scholar]
  20. Okamuto K., Mudd J.A., Marmur J. 1968; Conversion of Bacillus subtilis DNA to phage DNA following mitomycin C induction. Journal of Molecular Biology 34:429–437
    [Google Scholar]
  21. Sakaki Y., Karu A.E., Linn S., Echols H. 1973; Purification and properties of the #x00B7;protein specified by bacteriophage λ. An inhibitor of the host RecBC recombination enzyme. Proceedings of the National Academy ofSciences ofthe United States of America 70:2215–2219
    [Google Scholar]
  22. Sargent M.G., Bennett M.F. 1982; Attachment of the chromosomal terminus of Bacillus subtilis to a fast sedimenting complex. Journal of Bacreriology 150:623–632
    [Google Scholar]
  23. Sargent M.G., Bennett M. F. 1985; Amplification of a major membrane bound DNA sequence from Bacillus subtilis. Journal of Bacteriology 161:589–595
    [Google Scholar]
  24. Sargent M.G., Bennett M. F., Burdett I.J. 1983; Isolation of specific restriction fragments associated with a membrane sub-particle from Bacillus subrifis. Journal of Bacteriology 154:1389–1396
    [Google Scholar]
  25. Scher B., Dubnau D. 1973; A manganese stimulated endonuclease from Bacillus subtiis. Biochemical and Biophysical Research Communications595–602
    [Google Scholar]
  26. Strauss B., Marone R. 1967; A heat labile inhibitor of DNA degradation in Bacillus subtilis. Biochemical and Biophysical Research Communications 29:143–147
    [Google Scholar]
  27. Tanaka T. 1979; recE4-independent recombination between homologous DNA segments of Bacillus subtilis plasmids. Journal of Bacteriology 139:775–782
    [Google Scholar]
  28. Taylor A., Smith G.R. 1980; Unwinding and rewinding of DNA by the RecBC enzyme. Cell 22:447–457
    [Google Scholar]
  29. Trautner T.A., Pawlek B., Gunther U., Canosi S.U., Jentsch S., Freund M. 1980; Restriction and modification in Bacillus subrilis: identification of a gene in the temperate phage SPβ coding for a BsuR-specific modification methyltransferase. Molecular and General Genetics 180:361–367
    [Google Scholar]
  30. Uhlén M., Flock J., Phillipson L. 1981; RecE4-independent deletions of recombinant plasmids in Bacillus subtilis. Plasmid 5:161–169
    [Google Scholar]
  31. Van Vliet F., Coutuner M., De La Fonteyne J., Jedlicki E. 1978; Mu-l directed inhibition of DNA breakdown in Escherichia coli recA cells. Molecular and General Generics 164:109–112
    [Google Scholar]
  32. Devos W.M., Devries S.G., Venema G. 1983; Cloning and expression of the Escherichia coli recA gene in Bacillus subtilis. Gene 25:301–308
    [Google Scholar]
  33. Warner F.D., Ktros G.A., Romano M.P., Hemphill H.E. 1977; Characterisation of SPβ, a temperate bacteriophage from Bacillus subtilis Ml68. >Canadian Journal of Microbiology 23:45–51
    [Google Scholar]
  34. Wilson G.A., Borr K.F. 1968; Nutritional factors influencing the development of competence in the Bacillus subtilis transformation system. Journal of Bacteriology 95:1439–1449
    [Google Scholar]
  35. Yasbin R.E. 1977; DNA repair in Bacillus subtilis.II. Activation of the inducible system in competent bacteria. Molecular and General Genetics 153:219–225
    [Google Scholar]
  36. Yasbin R.E., Wilson G.A., Young F.E. 1975; Transformation and transfection in lysogenic strains of Bacillus subrilis: evidence for selective induction of prophage in competent cells. Journal of Bacteriology 121:296–304
    [Google Scholar]
  37. Young M. 1984; Gene amplification in Bacillus subrilis. Journal of General Microbiology 130:1613–1621
    [Google Scholar]
  38. Zahler S.A., Korman R.Z., Rosenthal R., Hemphill H.E. 1977; Bacillus subtilis bacteriophage SP β. Localisation of the prophageattachment site and specialised transduction. Journal of Bacteriaology 129:556–561
    [Google Scholar]
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