Skip to main content
Log in

Characterization of the transfer-related tra region of the conjugative plasmid p19 from a Bacillus subtilis soil strain

  • Genetics of Microorganisms
  • Published:
Russian Journal of Genetics Aims and scope Submit manuscript

Abstract

The nucleotide sequences of three DNA fragments (total size 30574 bp) of the plasmid p19 from the Bacillus subtilis 19 soil strain have been determined. Thirty open reading frames (ORFs) have been identified in these fragments. oriT of the plasmid has also been identified. As shown by the search for homologs of hypothetical protein products of these ORFs in databases, such homology exists for 18 ORFs. The protein products of nine ORFs can be assumed to have specific functions. Several ORFs were inactivated via insertional mutagenesis, and the conjugation capacity of the mutant plasmids was estimated. According to the data on homology of protein products and the results of ORF inactivation, regions of a total size of about 20 kb from the DNA fragments sequenced by us were inferred to belong to the tra region of p19. As follows from the analysis of the identified ORFs of the p19 tra region, it differs from the earlier described tra regions of other plasmids, irrespective of a certain similarity with the corresponding regions of plasmids of gram-positive bacteria from the genera Bacillus, Clostridium, and Listeria.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Koehler, T.M. and Thorne, C.B., Bacillus subtilis (natto) Plasmid pLS20 Mediates Interspecies Plasmid Transfer, J. Bacteriol., 1987, vol. 169, pp. 5271–5278.

    CAS  PubMed  Google Scholar 

  2. Itaya, M., Sakaya, N., Matsunaga, S., et al., Conjugational Transfer Kinetics of pLS20 between Bacillus subtilis in Liquid Medium, Biosci. Biotechnol. Biochem., 2005, vol. 70, pp. 740–742.

    Article  Google Scholar 

  3. Lotareva, O.V., Poluektova, E.U., Titok, M.A., and Prozorov, A.A., Large Plasmid from Bacillus subtilis Soil Strain, Performing Highly Frequent Conjugative Mobilization, Dokl. Akad. Nauk, 2001, vol. 379, no. 1, pp. 130–131.

    CAS  Google Scholar 

  4. Titok, M.A., Chapius, J., Selezneva, Y.V., et al., Bacillus subtilis Soil Isolates: Plasmid Replicon Analysis and Construction of a New Theta-Replicating Vector, Plasmid, 2003, vol. 49, no. 1, pp. 53–62.

    Article  CAS  PubMed  Google Scholar 

  5. Lotareva, O.V., Poluektova, E.U., Fedorina, E.A., et al., Interand Intraspesies Conjugal Transfer of Different Plasmids in Bacilli, Russ. J. Genet., 2003, vol. 39, no. 8, pp. 960–963.

    Article  CAS  Google Scholar 

  6. Poluektova, E.U., Fedorina, E.A., Lotareva, O.V., and Prozorov, A.A., Plasmid Transfer in Bacilli by a SelfTransmissible Plasmid p19 from a Bacillus subtilis Soil Strain, Plasmid, 2004, vol. 52, pp. 212–217.

    Article  CAS  PubMed  Google Scholar 

  7. Lotareva, O.V., Shilovskii, I.P., and Prozorov, A.A., A Phenomenon of Plasmid Retrotransfer in Conjugation of Bacillus subtilis, Russ. J. Genet., 2006, vol. 42, no. 12, pp. 1467–1469.

    Article  CAS  Google Scholar 

  8. Lotareva, O.V., Nezametdinova, V.Z., and Prozorov, A.A., Conjugative Chromosomal Gene Transfer in Bacillus subtilis, Russ. J. Genet., 2007, vol. 43, no. 7, pp. 742–747.

    Article  CAS  Google Scholar 

  9. Lotareva, O.V. and Prozorov, A.A., Conjugative Plasmid Transfer in Bacillus subtilis in Soil Microcosmos, Mikrobiologiya, 2003, vol. 72, no. 6, pp. 780–784.

    CAS  Google Scholar 

  10. Lotareva, O.V. and Prozorov, A.A., Conjugation in Bacillus subtilis in Soils of Natural Landscapes, Mikrobiologiya, 2005, vol. 74, no. 1, pp. 132–134.

    CAS  Google Scholar 

  11. Poluektova, E.U., Gagarina, E.Yu., Shilovskii, I.P., et al., Molecular Analysis of Some Genes from Plasmid p19 of the Bacillus subtilis 19 Soil Strain Involved in Conjugation, Russ. J. Genet., 2008, vol. 44, no. 5, pp. 539–545.

    Article  CAS  Google Scholar 

  12. Bullock, W.O., Fernandez, J.M., and Short, M.J., X-L1blue, a High Efficiency Plasmid Transforming recA Escherichia coli Strain with Beta Galactosidase Selection, BioTechniques, 1987, vol. 5, pp. 376–379.

    CAS  Google Scholar 

  13. Chambers, S.P., Prior, S.E., Barstow, D.A., and Minton, N.P., The pMTL Cloning Vectors. I. Improved Polylinker Regions to Facilitate the Generation of Sonicated DNA for Nucleotide Sequence, Gene, 1988, vol. 68, pp. 139–149.

    Article  CAS  PubMed  Google Scholar 

  14. Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Lab., 1989.

    Google Scholar 

  15. Prozorov, A.A., Transformatsiya u bakterii (Transformation in Bacteria), Moscow: Nauka, 1988.

    Google Scholar 

  16. Sangar, F., Nicklen, S., and Coulson, A.R., DNA Sequencing with Chain-Terminating Inhibitors, Proc. Natl. Acad. Sci. USA, 1977, vol. 74, pp. 5463–5467.

    Article  Google Scholar 

  17. Helmann, J.D., Compilation and Analysis of Bacillus subtilis Sigma A-Dependent Promoter Sequences: Evidence for Extended Contact between RNA Polymerase and Upstream Promoter DNA, Nucleic Acids Res., 1995, vol. 23, pp. 2351–2360.

    Article  CAS  PubMed  Google Scholar 

  18. Zechner, E.L, de la Cruz, F., Eisenbrandt, R., et al., Conjugative DNA Transfer Processes, The Horizontal Gene Pool, Thomas, C.M., Ed., Amsterdam: Harwood Academic, 2000, pp. 87–174.

    Google Scholar 

  19. Schroder, S. and Lanka, E., The Mating Pair Formation System of Conjugative Plasmids-A Versatile Secretion Machinery for Transfer of Proteins and DNA, Plasmid, 2005, vol. 54, no. 1, pp. 1–25.

    Article  PubMed  CAS  Google Scholar 

  20. Grohmann, E., Muth, G., and Espinosa, M., Conjugative Plasmid Transfer in Gram-Positive Bacteria, Microbiol. Mol. Biol. Rev., 2003, vol. 67, pp. 277–301.

    Article  CAS  PubMed  Google Scholar 

  21. Atmakuri, K., Cascales, E., and Christie, P.J., Energetic Components VirD4, VirB11 and VirB4 Mediate Early Transfer Reactions Required for Bacterial Type IV Secretion, Mol. Microbiol., 2004, vol. 54, no. 5, pp. 1199–1211.

    Article  CAS  PubMed  Google Scholar 

  22. Teng, W.L., Bannam, T.L., Parsons, J.A., and Rood, J.I., Functional Characterization and Localization of the TcpH Conjugation Protein from Clostridium perfringens, J. Bacteriol., 2008, vol. 190, no. 14, pp. 5075–5086.

    Article  CAS  PubMed  Google Scholar 

  23. Tomita, H. and Ike, Y., Genetic Analysis of TransferRelated Regions of the Vancomicin Resistance Enterococcus Conjugative Plasmid pHTβ: Identification of oriT and Putative Relaxase Gene, J. Bacteriol., 2005, vol. 187, no. 22, pp. 7727–7737.

    Article  CAS  PubMed  Google Scholar 

  24. Francia, M.V., Varsaki, A., Garcillan-Barcia, M.P., et al., A Classification Scheme for Mobilization Regions of Bacterial Plasmids, FEMS Microbiol. Rev., 2004, vol. 28, pp. 79–100.

    Article  CAS  PubMed  Google Scholar 

  25. Lagodich, A.V., Shtanyuk, Ya.V., Prozorov, A.A., and Titok, M.A., Characteristics of the Plasmid Replication System from the Natural Bacillus subtilis Strains, Mol. Biol. (Moscow), 2004, vol. 38, no. 3, pp. 437–441.

    CAS  Google Scholar 

  26. Nezametdinova, V.Z., Fedorina, E.A., Poluektova, E.U., et al., Conjugative Ability and Comparative Characteristic of Large Plasmids from Natural Strains of Bacillus subtilis from Different Regions of East European Plain, Mikrobiologiya, 2007, vol. 76, no. 2, pp. 215–222.

    Google Scholar 

  27. Van der Auwera, G.A., Andrup, L., and Mahillon, J., Conjugative Plasmid pAW63 Brings New Insights into the Genesis of the Bacillus anthracis Virulence Plasmid pXO2 and of the Bacillus thuringiensis Plasmid pBT9727, BMC Genomic, 2005, vol. 6, no. 103.

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to E. U. Poluektova.

Additional information

Original Russian Text © E.U. Poluektova, E.Yu. Gagarina, V.Z. Nezametdinova, I.P. Chilovskii, S.A. Rodionova, A.A. Prozorov, 2010, published in Genetika, 2010, Vol. 46, No. 1, pp. 33–43.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Poluektova, E.U., Gagarina, E.Y., Nezametdinova, V.Z. et al. Characterization of the transfer-related tra region of the conjugative plasmid p19 from a Bacillus subtilis soil strain. Russ J Genet 46, 26–35 (2010). https://doi.org/10.1134/S1022795410010059

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1022795410010059

Keywords

Navigation