Use of Integrational Plasmid Vectors to Demonstrate the Polycistronic Nature of a Transcriptional Unit () Required for Sporulation of Free

Abstract

Plasmids carrying different portions of the polycistronic locus, and unable to replicate autonomously in , were able to transform a Spo strain, 151, for the plasmid-determined chloramphenicol resistance by Campbell-like insertion into the region of homology on the chromosome. Two such plasmids, p35 and p36, yielded Spo-transformants, indicating that the cloned regions of these plasmids were entirely within the chromosomal transcriptional unit. The cloned regions overlapped the end of a known cistron, so that the transcriptional unit was larger than this cistron, and was polycistronic. This is the first demonstration of such a polycistronic sporulation transcriptional unit. The sequence of the region has now been determined (given in an accompanying paper) and suggests a transcriptional unit with three open reading frames. Two other plasmids yielded Spo transformants of 151, and these define the outer limits of the transcriptional unit. The adjacent sporulation locus identified by the mutation was not part of the same transcriptional unit.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-130-8-2123
1984-08-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/130/8/mic-130-8-2123.html?itemId=/content/journal/micro/10.1099/00221287-130-8-2123&mimeType=html&fmt=ahah

References

  1. Bott K. F., Wilson G. A. 1968; Metabolic and nutritional factors influencing the development of competence for transfection of Bacillus subtilis. Bacteriological Reviews 37:370–378
    [Google Scholar]
  2. Brown M. G. M, Watson A., Saunders J. R., Humphreys G. O. 1979; Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth. FEMS Microbiology Letters 5:219–222
    [Google Scholar]
  3. Canosi V., Iglesias A., Trautner T. A. 1981; Plasmid transformation in Bacillus subtilis: effects of insertion of Bacillus subtilis DNA into plasmid pC 194. Molecular and General Genetics 181:434–440
    [Google Scholar]
  4. Chak K.-F., Lencastre H.DE, Piggot P. J. 1982; Facile in vivo transfer of mutations between the Bacillus subtilis chromosome and a plasmid harbouring homologous DNA. Journal of General Microbiology 128:2813–2816
    [Google Scholar]
  5. Davis R. W., Botstein D., Roth J. R. 1980 Advanced Bacterial Genetics Cold Spring Harbor, New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  6. 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]
  7. Errington J., Mandelstam J. 1983; Variety of sporulation phenotypes resulting from mutations in a single regulatory locus, spoil A, in Bacillus subtilis. Journal of General Microbiology 129:2091–2101
    [Google Scholar]
  8. Ferrari F. A., Nguyen A., Lang D., Hoch J. A. 1983; Construction and properties of an integrable plasmid for Bacillus subtilis. Journal of Bacteriology 154:1513–1515
    [Google Scholar]
  9. Fort P., Piggot P. J. 1984; Nucleotide sequence of sporulation locus spoIIA in Bacillus subtilis. Journal of General Microbiology 130:2147–2153
    [Google Scholar]
  10. Guerry P., Leblanc D. J., Falkow S. 1973; General method for the isolation of plasmid deoxyribonucleic acid. Journal of Bacteriology 116:1064–1066
    [Google Scholar]
  11. Gutterson N. I., Koshland D. E. 1983; Replacement and amplification of bacterial genes with sequences altered in vitro. Proceedings of the National Academy of Sciences of the United States of America 80:4894–4898
    [Google Scholar]
  12. Haldenwang W. G., Banner C. D. B, Ollington J. F., Losick , Hoch J. A., O’Connor M. B., Sonenshein A. L. 1980; Mapping a cloned gene under sporulation control by insertion of a drug resistance marker into the Bacillus subtilis chromosome. Journal of Bacteriology 142:90–98
    [Google Scholar]
  13. Hranueli D., Piggot P. J., Mandelstam J. 1974; Statistical estimate of the total number of operons specific for Bacillus subtilis sporulation. Journal of Bacteriology 119:684–690
    [Google Scholar]
  14. Iglesias A., Bensi G., Canosi U., Trautner T. A. 1981; Plasmid transformation in Bacillus subtilis. Alterations introduced into the recipient-homologous DNA of hybrid plasmids can be corrected in transformation. Molecular and General Genetics 184:405–409
    [Google Scholar]
  15. Jeffreys A. J., Flavell R. A. 1977; A physical map of the DNA regions flanking the rabbit β-globin gene. Cell 12:429–439
    [Google Scholar]
  16. Lencastre H.DE, Piggot P. J. 1983; Use of the Escherichia coli transposon Tn 1000 (yd) to generate mutations in Bacillus subtilis DNA. Journal of General Microbiology 129:3203–3210
    [Google Scholar]
  17. Liu H.-M., Chak K.-F., Piggot P. J. 1982; Isolation and characterization of a recombinant plasmid carrying a functional part of the Bacillus subtilis spoil A locus. Journal of General Microbiology 128:2805–2812
    [Google Scholar]
  18. Mandelstam J. 1976; Bacterial sporulation: a problem in the biochemistry and genetics of a primitive developmental system. Proceedings of the Royal Society B193:89–106
    [Google Scholar]
  19. Niaudet B., Goze A., Ehrlich S. D. 1982; Insertional mutagenesis in Bacillus subtilis: mechanism and use in gene cloning. Gene 19:277–284
    [Google Scholar]
  20. Piggot P. J. 1973; Mapping of asporogenous mutations of Bacillus subtilis: a minimum estimate of the number of sporulation operons. Journal of Bacteriology 114:1241–1253
    [Google Scholar]
  21. Piggot P. J. 1979; Genetic strategies for studying bacterial differentiation. Biological Reviews 54:347–367
    [Google Scholar]
  22. Piggot P. J., Coote J. G. 1976; Genetic aspects of bacterial endospore formation. Bacteriological Reviews 40:908–962
    [Google Scholar]
  23. Piggot P. J., Moir A., Smith D. A. 1981; Advances in the genetics of Bacillus subtilis differentiation. In Spores VIII pp. 29–39 Edited by Levinson H., Tipper D., Sonenshein A. L. Washington, D.C.: American Society for Microbiology;
    [Google Scholar]
  24. Primrose S. B., Ehrlich S. D. 1981; Isolation of plasmid deletion mutants and study of their instability. Plasmid 6:193–201
    [Google Scholar]
  25. Roberts T. M., Swanberg S. L., Poteete A., Riedel G., Backman K. 1980; A plasmid cloning vehicle allowing a positive selection for inserted fragments. Gene 12:123–127
    [Google Scholar]
  26. Savva D., Mandelstam J. 1984; Cloning of the Bacillus subtilis spoil A and spoVA loci in phage ø105DI:It. Journal of General Microbiology 130:2137–2145
    [Google Scholar]
  27. Southern E. 1979; Gel electrophoresis of restriction fragments. Methods of Enzymology 68:152–176
    [Google Scholar]
  28. Struhl K. 1983; The new yeast genetics. Nature, London 305:391–397
    [Google Scholar]
  29. Waites W. M., Kay D., Dawes I. W., Wood D. A., Warren S. C., Mandelstam J. 1970; Sporula- tion in Bacillus subtilis. Correlation of biochemical events with morphological changes in asporogenous mutants. Biochemical Journal 118:667–676
    [Google Scholar]
  30. Wilson F. E., Hoch J. A., Bott K. 1981; Genetic mapping of a linked cluster of ribosomal ribonucleic acid genes in Bacillus subtilis. Journal of Bacteriology 148:624–628
    [Google Scholar]
  31. Young F. E., Wilson G. A. 1974; Bacillus subtilis. In Handbook of Genetics 1: pp. 69–114
    [Google Scholar]
  32. Young M. 1983; The mechanism of insertion of a segment of heterologous DN A into the chromosome of Bacillus subtilis. Journal of General Microbiology 129:1497–1512
    [Google Scholar]
  33. Yudkin M. D., Turley L. 1981; Mapping of six mutations in the spoil A locus of Bacillus subtilis and studies of their response to a nonsense suppressor. Journal of General Microbiology 124:255–261
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-130-8-2123
Loading
/content/journal/micro/10.1099/00221287-130-8-2123
Loading

Data & Media loading...

Most cited Most Cited RSS feed