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. 1978 Mar;75(3):1423–1427. doi: 10.1073/pnas.75.3.1423

Molecular cloning of genetically active fragments of Bacillus DNA in Bacillus subtilis and properties of the vector plasmid pUB110.

K M Keggins, P S Lovett, E J Duvall
PMCID: PMC411484  PMID: 418411

Abstract

Plasmid pUB110 (approximately 2.8 x 10(6) daltons), originally detected in Staphylococcus aureus, specifies resistance to neomycin and has been transformed into Bacillus subtilis 168, In B. subtilis, pUB110 is stably maintained at about 50 copies per chromosome and renders the host resistant to neomycin sulfate at 5 microgram/ml. pUB110 isolated from B. subtilis transforms Rec+ and recE4-containing strains of B. subtilis at frequencies greater than or equal to 10(3) transformants per microgram of plasmid. pUB110 was transferred by PBS1 or SP10 transduction from B. subtilis to strains of B. pumilus and B. licheniformis. pUB110 is compatible with each of four previously described Bacillus plasmids, including pPL576, pPL10, pPL7065, and pPL2. pUB110 contains a single EcoR1-sensitive site and was used as vector to clone DNA fragments that complement the trpC2 mutation in B. subtilis 168 from EcoR1 digests of the chromosome DNA isolated from B. pumilus strains NRRL B-3275 and NRS576, B. licheniformis strains 9945A and 749C, and B. subtilis 168. Genetic and physical properties of each of the constructed Trp derivatives of pUB110 are described.

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Selected References

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

  1. Anagnostopoulos C., Crawford I. P. Le groupe des gènes régissant la biosynthèse du tryptophane chez Bacillus subtilis. C R Acad Sci Hebd Seances Acad Sci D. 1967 Jul 3;265(1):93–96. [PubMed] [Google Scholar]
  2. Bott K. F., Wilson G. A. Development of competence in the Bacillus subtilis transformation system. J Bacteriol. 1967 Sep;94(3):562–570. doi: 10.1128/jb.94.3.562-570.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bramucci M. G., Lovett P. S. Low-frequency, pbsi-mediated plasmid transduction in Bacillus pumilus. J Bacteriol. 1976 Aug;127(2):829–831. doi: 10.1128/jb.127.2.829-831.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clarke L., Carbon J. Biochemical construction and selection of hybrid plasmids containing specific segments of the Escherichia coli genome. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4361–4365. doi: 10.1073/pnas.72.11.4361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clowes R. C. Molecular structure of bacterial plasmids. Bacteriol Rev. 1972 Sep;36(3):361–405. doi: 10.1128/br.36.3.361-405.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen S. N., Chang A. C., Boyer H. W., Helling R. B. Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240–3244. doi: 10.1073/pnas.70.11.3240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Dubnau D., Cirigliano C. Genetic characterization of recombination-deficient mutants of Bacillus subtilis. J Bacteriol. 1974 Feb;117(2):488–493. doi: 10.1128/jb.117.2.488-493.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dubnau D., Smith I., Morell P., Marmur J. Gene conservation in Bacillus species. I. Conserved genetic and nucleic acid base sequence homologies. Proc Natl Acad Sci U S A. 1965 Aug;54(2):491–498. doi: 10.1073/pnas.54.2.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ehrlich S. D. Replication and expression of plasmids from Staphylococcus aureus in Bacillus subtilis. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1680–1682. doi: 10.1073/pnas.74.4.1680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldberg I. D., Gwinn D. D., Thorne C. B. Interspecies transformation between Bacillus subtilis and Bacillus licheniformis. Biochem Biophys Res Commun. 1966 May 25;23(4):543–548. [PubMed] [Google Scholar]
  12. Hoch S. O., Anagnostopoulos C., Crawford I. P. Enzymes of the tryptophan operon of Bacillus subtilis. Biochem Biophys Res Commun. 1969 Jun 27;35(6):838–844. doi: 10.1016/0006-291x(69)90700-1. [DOI] [PubMed] [Google Scholar]
  13. Klotz L. C., Zimm B. H. Size of DNA determined by viscoelastic measurements: results on bacteriophages, Bacillus subtilis and Escherichia coli. J Mol Biol. 1972 Dec 30;72(3):779–800. doi: 10.1016/0022-2836(72)90191-x. [DOI] [PubMed] [Google Scholar]
  14. Lovett P. S., Bramucci M. G. Plasmid deoxyribonucleic acid in Bacillus subtilis and Bacillus pumilus. J Bacteriol. 1975 Oct;124(1):484–490. doi: 10.1128/jb.124.1.484-490.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lovett P. S., Duvall E. J., Bramucci M. G., Taylor R. Host function specified by Bacillus pumilus plasmid pPL7065. Antimicrob Agents Chemother. 1977 Sep;12(3):435–437. doi: 10.1128/aac.12.3.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lovett P. S., Duvall E. J., Keggins K. M. Bacillus pumilus plasmid pPL10: properties and insertion into Bacillus subtilis 168 by transformation. J Bacteriol. 1976 Aug;127(2):817–828. doi: 10.1128/jb.127.2.817-828.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lovett P. S. Plasmid in Bacillus pumilus and the enhanced sporulation of plasmid-negative variants. J Bacteriol. 1973 Jul;115(1):291–298. doi: 10.1128/jb.115.1.291-298.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lovett P. S., Young F. E. Genetic analysis in Bacillus pumilus by PBSI-mediated transduction. J Bacteriol. 1970 Feb;101(2):603–608. doi: 10.1128/jb.101.2.603-608.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lovett P. S., Young F. E. Identification of Bacillus subtilis NRRL B-3275 as a strain of Bacillus pumilus. J Bacteriol. 1969 Nov;100(2):658–661. doi: 10.1128/jb.100.2.658-661.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mahler I., Halvorson H. O. Transformation of Escherichia coli and Bacillus subtilis with a hybrid plasmid molecule. J Bacteriol. 1977 Jul;131(1):374–377. doi: 10.1128/jb.131.1.374-377.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. STUDIER F. W. SEDIMENTATION STUDIES OF THE SIZE AND SHAPE OF DNA. J Mol Biol. 1965 Feb;11:373–390. doi: 10.1016/s0022-2836(65)80064-x. [DOI] [PubMed] [Google Scholar]
  22. Spizizen J. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE. Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072–1078. doi: 10.1073/pnas.44.10.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. TAYLOR M. J., THORNE C. B. TRANSDUCTION OF BACILLUS LICHENIFORMIS AND BACILLUS SUBTILIS BY EACH OF TWO PHAGES. J Bacteriol. 1963 Sep;86:452–461. doi: 10.1128/jb.86.3.452-461.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tanaka T., Koshikawa T. Isolation and characterization of four types of plasmids from Bacillus subtilis (natto). J Bacteriol. 1977 Aug;131(2):699–701. doi: 10.1128/jb.131.2.699-701.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tanaka T., Kuroda M., Sakaguchi K. Isolation and characterization of four plasmids from Bacillus subtilis. J Bacteriol. 1977 Mar;129(3):1487–1494. doi: 10.1128/jb.129.3.1487-1494.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Thomas M., Davis R. W. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. doi: 10.1016/0022-2836(75)90383-6. [DOI] [PubMed] [Google Scholar]
  27. Tyeryar F. J., Jr, Lawton W. D., MacQuillan A. M. Sequential replication of the chromosome of Bacillus licheniformis. J Bacteriol. 1968 Jun;95(6):2062–2069. doi: 10.1128/jb.95.6.2062-2069.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tyeryar F. J., Jr, Taylor M. J., Lawton W. D., Goldberg I. D. Cotransduction and cotransformation of genetic markers in Bacillus subtilis and Bacillus licheniformis. J Bacteriol. 1969 Nov;100(2):1027–1036. doi: 10.1128/jb.100.2.1027-1036.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wilson G. A., Young F. E. Intergenotic transformation of the Bacillus subtilis genospecies. J Bacteriol. 1972 Sep;111(3):705–716. doi: 10.1128/jb.111.3.705-716.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]

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