Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1989 Feb;171(2):1166–1172. doi: 10.1128/jb.171.2.1166-1172.1989

Replication and segregational stability of Bacillus plasmid pBAA1.

K M Devine 1, S T Hogan 1, D G Higgins 1, D J McConnell 1
PMCID: PMC209715  PMID: 2492507

Abstract

A cryptic plasmid, pBAA1, was identified in an industrial Bacillus strain. The plasmid is 6.8 kilobases in size and is present in cells at a copy number of approximately 5 per chromosome equivalent. The plasmid has been maintained under industrial fermentation conditions without apparent selective pressure and so is assumed to be partition proficient. The minimal replicon was localized to a 1.4-kilobase fragment which also contains the functions required for copy number control. The very low level of segregational instability of the minimal replicon suggests that it also contains functions involved in plasmid maintenance. Comparison with other plasmids indicates that pBAA1 belongs to the group of small gram-positive plasmids which replicate by a rolling cycle-type mechanism. A sequence was identified which is required for the efficient conversion of the single plus strand to the double-stranded form during plasmid replication. Deletion of this sequence resulted in a low level of segregational plasmid instability.

Full text

PDF
1168

Images in this article

Selected References

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

  1. Bron S., Bosma P., van Belkum M., Luxen E. Stability function in the Bacillus subtilis plasmid pTA 1060. Plasmid. 1987 Jul;18(1):8–15. doi: 10.1016/0147-619x(87)90073-4. [DOI] [PubMed] [Google Scholar]
  2. Bron S., Luxen E. Segregational instability of pUB110-derived recombinant plasmids in Bacillus subtilis. Plasmid. 1985 Nov;14(3):235–244. doi: 10.1016/0147-619x(85)90007-1. [DOI] [PubMed] [Google Scholar]
  3. Bron S., Venema G. Ultraviolet inactivation and excision-repair in Bacillus subtilis. I. Construction and characterization of a transformable eightfold auxotrophic strain and two ultraviolet-sensitive derivatives. Mutat Res. 1972 May;15(1):1–10. doi: 10.1016/0027-5107(72)90086-3. [DOI] [PubMed] [Google Scholar]
  4. Chang S., Chang S. Y., Gray O. Structural and genetic analyses of a par locus that regulates plasmid partition in Bacillus subtilis. J Bacteriol. 1987 Sep;169(9):3952–3962. doi: 10.1128/jb.169.9.3952-3962.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ehrlich S. D. DNA cloning in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1433–1436. doi: 10.1073/pnas.75.3.1433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Fiddes J. C., Barrell B. G., Godson G. N. Nucleotide sequences of the separate origins of synthesis of bacteriophage G4 viral and complementary DNA strands. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1081–1085. doi: 10.1073/pnas.75.3.1081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grandi G., Mottes M., Sgaramella V. Specific pattern of instability of Escherichia coli HisG gene cloned in Bacillus subtilis via the Staphylococcus aureus plasmid pCS194. Plasmid. 1981 Jul;6(1):99–111. doi: 10.1016/0147-619x(81)90057-3. [DOI] [PubMed] [Google Scholar]
  9. Gros M. F., te Riele H., Ehrlich S. D. Rolling circle replication of single-stranded DNA plasmid pC194. EMBO J. 1987 Dec 1;6(12):3863–3869. doi: 10.1002/j.1460-2075.1987.tb02724.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gruss A. D., Ross H. F., Novick R. P. Functional analysis of a palindromic sequence required for normal replication of several staphylococcal plasmids. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2165–2169. doi: 10.1073/pnas.84.8.2165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Gryczan T. J., Dubnau D. Construction and properties of chimeric plasmids in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1428–1432. doi: 10.1073/pnas.75.3.1428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gryczan T., Shivakumar A. G., Dubnau D. Characterization of chimeric plasmid cloning vehicles in Bacillus subtilis. J Bacteriol. 1980 Jan;141(1):246–253. doi: 10.1128/jb.141.1.246-253.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Khan S. A., Adler G. K., Novick R. P. Functional origin of replication of pT181 plasmid DNA is contained within a 168-base-pair segment. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4580–4584. doi: 10.1073/pnas.79.15.4580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kieny M. P., Lathe R., Lecocq J. P. New versatile cloning and sequencing vectors based on bacteriophage M13. Gene. 1983 Dec;26(1):91–99. doi: 10.1016/0378-1119(83)90039-2. [DOI] [PubMed] [Google Scholar]
  16. Lopez P., Espinosa M., Greenberg B., Lacks S. A. Generation of deletions in pneumococcal mal genes cloned in Bacillus subtilis. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5189–5193. doi: 10.1073/pnas.81.16.5189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Murai M., Miyashita H., Araki H., Seki T., Oshima Y. Molecular structure of the replication origin of a Bacillus amyloliquefaciens plasmid pFTB14. Mol Gen Genet. 1987 Nov;210(1):92–100. doi: 10.1007/BF00337763. [DOI] [PubMed] [Google Scholar]
  18. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  19. Novick R. P., Adler G. K., Majumder S., Khan S. A., Carleton S., Rosenblum W. D., Iordanescu S. Coding sequence for the pT181 repC product: a plasmid-coded protein uniquely required for replication. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4108–4112. doi: 10.1073/pnas.79.13.4108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ortlepp S. A., Ollington J. F., McConnell D. J. Molecular cloning in Bacillus subtilis of a Bacillus licheniformis gene encoding a thermostable alpha amylase. Gene. 1983 Sep;23(3):267–276. doi: 10.1016/0378-1119(83)90017-3. [DOI] [PubMed] [Google Scholar]
  21. Ostroff G. R., Pène J. J. Molecular cloning with bifunctional plasmid vectors in Bacillus subtilis. II. Transfer of sequences propagated in Escherichia coli to B. subtilis. Mol Gen Genet. 1984;193(2):306–311. doi: 10.1007/BF00330685. [DOI] [PubMed] [Google Scholar]
  22. Rabinovich P. M., Haykinson MYa, Arutyunova L. S., Yomantas YuV, Stepanov A. I. The structure and source of plasmid DNA determine the cloning properties of vectors for Bacillus subtilis. Basic Life Sci. 1985;30:635–656. doi: 10.1007/978-1-4613-2447-8_44. [DOI] [PubMed] [Google Scholar]
  23. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shlomai J., Kornberg A. An Escherichia coli replication protein that recognizes a unique sequence within a hairpin region in phi X174 DNA. Proc Natl Acad Sci U S A. 1980 Feb;77(2):799–803. doi: 10.1073/pnas.77.2.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  26. del Solar G. H., Puyet A., Espinosa M. Initiation signals for the conversion of single stranded to double stranded DNA forms in the streptococcal plasmid pLS1. Nucleic Acids Res. 1987 Jul 24;15(14):5561–5580. doi: 10.1093/nar/15.14.5561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. te Riele H., Michel B., Ehrlich S. D. Single-stranded plasmid DNA in Bacillus subtilis and Staphylococcus aureus. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2541–2545. doi: 10.1073/pnas.83.8.2541. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES