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. 1984 May;158(2):507–512. doi: 10.1128/jb.158.2.507-512.1984

Cloning of sporulation gene spoIIG in Bacillus subtilis.

H Ayaki, Y Kobayashi
PMCID: PMC215457  PMID: 6327608

Abstract

Two specialized transducing phages carrying a sporulation gene, spoIIG , of Bacillus subtilis were constructed from B. subtilis temperate phages p11 and phi 105 by the "prophage transformation" method. Restriction enzyme analysis and transformation experiments showed that the spoIIG gene was present on a 6.2 X 10(6)-dalton (6.2-Md) EcoRI fragment in both transducing phage genomes. Further analysis showed that spoIIG + transforming activity resides on a 2.25-Md EcoRI-BamHI fragment within the 6.2-Md EcoRI fragment. The 2.25-Md fragment was subcloned into the region between the EcoRI and BamHI sites of pUB110, and deletion plasmids lacking PstI or HindIII fragments within the 2.25-Md fragment were constructed. The recombinant plasmid carrying the intact spoIIG gene restored sporulation of strain HU1002 ( spoIIG41 recE4 ) to a frequency of 10(4) spores per ml and inhibited sporulation of strain 4309 ( spo + recE4 ) to a level of 10(3) spores per ml.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bonamy C., Szulmajster J. Cloning and expression of Bacillus subtilis spore genes. Mol Gen Genet. 1982;188(2):202–210. doi: 10.1007/BF00332676. [DOI] [PubMed] [Google Scholar]
  2. Dean D. H., Orrego J. C., Hutchison K. W., Halvorson H. O. New temperate bacteriophage for Bacillus subtilis, rho 11. J Virol. 1976 Nov;20(2):509–519. doi: 10.1128/jvi.20.2.509-519.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ferrari E., Henner D. J., Hoch J. A. Isolation of Bacillus subtilis genes from a charon 4A library. J Bacteriol. 1981 Apr;146(1):430–432. doi: 10.1128/jb.146.1.430-432.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gryczan T. J., Contente S., Dubnau D. Characterization of Staphylococcus aureus plasmids introduced by transformation into Bacillus subtilis. J Bacteriol. 1978 Apr;134(1):318–329. doi: 10.1128/jb.134.1.318-329.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Henner D. J., Hoch J. A. The Bacillus subtilis chromosome. Microbiol Rev. 1980 Mar;44(1):57–82. doi: 10.1128/mr.44.1.57-82.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hirochika H., Kobayashi Y., Kawamura F., Saito H. Cloning of sporulation gene spoOB of Bacillus subtilis and its genetic and biochemical analysis. J Bacteriol. 1981 May;146(2):494–505. doi: 10.1128/jb.146.2.494-505.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hirochika H., Kobayashi Y. Suppression of temperature-sensitive sporulation of a Bacillus subtilis elongation factor G mutant by RNA polymerase mutations. J Bacteriol. 1978 Dec;136(3):883–893. [PMC free article] [PubMed] [Google Scholar]
  8. Hoch J. A. Selection of cells transformed to prototrophy for sporulation markers. J Bacteriol. 1971 Mar;105(3):1200–1201. doi: 10.1128/jb.105.3.1200-1201.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jalanko A., Palva I., Söderlund Restriction maps of plasmids pUB110 and pBD9. Gene. 1981 Sep;14(4):325–328. doi: 10.1016/0378-1119(81)90165-7. [DOI] [PubMed] [Google Scholar]
  11. Kawamura F., Saito H., Ikeda Y. A method for construction of specialized transducing phage rho 11 of Bacillus subtilis. Gene. 1979 Feb;5(2):87–91. doi: 10.1016/0378-1119(79)90095-7. [DOI] [PubMed] [Google Scholar]
  12. Liu H. M., Chak K. F., Piggot P. J. Isolation and characterization of a recombinant plasmid carrying a functional part of the Bacillus subtilis spoIIA locus. J Gen Microbiol. 1982 Nov;128(11):2805–2812. doi: 10.1099/00221287-128-11-2805. [DOI] [PubMed] [Google Scholar]
  13. Loening U. E. Molecular weights of ribosomal RNA in relation to evolution. J Mol Biol. 1968 Dec;38(3):355–365. doi: 10.1016/0022-2836(68)90391-4. [DOI] [PubMed] [Google Scholar]
  14. Okamoto K., Mudd J. A., Mangan J., Huang W. M., Subbaiah T. V., Marmur J. Properties of the defective phage of Bacillus subtilis. J Mol Biol. 1968 Jun 28;34(3):413–428. doi: 10.1016/0022-2836(68)90169-1. [DOI] [PubMed] [Google Scholar]
  15. Rutberg L. Mapping of a temperate bacteriophage active on Bacillus subtilis. J Virol. 1969 Jan;3(1):38–44. doi: 10.1128/jvi.3.1.38-44.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shibata T., Ando T. Host controlled modification and restriction in Bacillus subtilis. Mol Gen Genet. 1974;131(4):275–280. doi: 10.1007/BF00264858. [DOI] [PubMed] [Google Scholar]
  17. Smith H. O. Recovery of DNA from gels. Methods Enzymol. 1980;65(1):371–380. doi: 10.1016/s0076-6879(80)65048-4. [DOI] [PubMed] [Google Scholar]
  18. Wilson G. A., Bott K. F. Nutritional factors influencing the development of competence in the Bacillus subtilis transformation system. J Bacteriol. 1968 Apr;95(4):1439–1449. doi: 10.1128/jb.95.4.1439-1449.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Yang R., Lis J., Wu R. Elution of DNA from agarose gels after electrophoresis. Methods Enzymol. 1979;68:176–182. doi: 10.1016/0076-6879(79)68012-6. [DOI] [PubMed] [Google Scholar]
  20. Young M. Use of temperature-sensitive mutants to study gene expression during sporulation in Bacillus subtilis. J Bacteriol. 1976 May;126(2):928–936. doi: 10.1128/jb.126.2.928-936.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

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