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Characterization of the lactococcal conjugative element pRS01 using IS946-mediated mutagenesis

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Methods for studying the genetics, molecular biology, physiology, and pathogenesis of the streptococci

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Abstract

Conjugation is a common mode of genetic transfer among the lactic acid bacteria. In an effort to exploit conjugation as a means of lactococcal strain development, we have characterized the transfer regions of pRS01 by insertional mutagenesis via IS946-mediated cointegration with the lactococcal-Escherichia coli shuttle plasmid pTRK28. In this work we will discuss our use of pTRK28-mediated cointegration to identify, clone, and characterize the pRS01 transfer regions including the conjugative origin of transfer.

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Abbreviations

Cmr :

= resistant to chloramphenicol

Ermr :

= resistant to erythromycin

IS:

= insertion sequence

LAB:

= lactic acid bacteria

oriT :

= origin of transfer

PBS:

= phosphate-buffered saline

Rec+ :

= recombination proficient

Rifr :

= resistant to rifampicin

Spcr :

= resistant to spectinomycin

Strr :

= resistant to streptomycin

References

  1. Anderson DG, McKay LL (1984). Genetic and physical characterization of recombinant plasmids associated with cell aggregation and high-frequency conjugal transfer in Streptococcus lactis ML3. J Bacteriol 158: 954–962.

    PubMed  CAS  Google Scholar 

  2. Ausubel FM, Brent R, Kingston RE, et al. (1995). Current protocols in molecular biology. New York: Wiley.

    Google Scholar 

  3. Behnke D, Gilmore MS, Ferretti J (1981). Plasmid pGB301, a new multiple resistance streptococcal cloning vehicle and its use in cloning of the gentamycin/kanamycin resistance determinant. Mol Gen Genet 182: 414–421.

    Article  PubMed  CAS  Google Scholar 

  4. Dower WJ, Miller JF, Ragsdale CW (1988). High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acid Res 16: 6127–6145.

    Article  PubMed  CAS  Google Scholar 

  5. Holo H, Nes IF (1995). Transformation of Lactococcus by electroporation. Methods Mol Biol 47: 195–199.

    PubMed  CAS  Google Scholar 

  6. Langella P, Le LY, Ehrlich SD, Gruss A (1993). Efficient plasmid mobilization by pIP501 in Lactococcus lactis subsp. lactis. J Bacteriol 175: 5806–5813.

    PubMed  CAS  Google Scholar 

  7. Lanka E, Wilkins BM (1995). DNA processing reactions in bacterial conjugation. Ann Rev Biochem 64: 141–169.

    Article  PubMed  CAS  Google Scholar 

  8. Mills DA, Choi CK, Dunny GM, McKay LL (1994). Genetic analysis of regions of the Lactococcus lactis subsp. lactis plasmid pRS01 involved in conjugative transfer. Appl Environ Microbiol 60: 4413–4420.

    PubMed  CAS  Google Scholar 

  9. O’Sullivan DJ, Klaenhammer TR (1993). Rapid mini-prep isolation of high-quality plasmid DNA from Lactococcus and Lactobacillus spp. Appl Environ Microbiol 59: 2730–2733.

    PubMed  Google Scholar 

  10. Phister TG, Mills DA, Cocconcelli PS, McKay LL (1996). Host range analysis of lactococcal conjugal element pRS01. In: Abstracts of the 96th General Meeting of the American Society for Microbiology, abstr. H-90, p 498. Washington, DC: American Society for Microbiology.

    Google Scholar 

  11. Reimmann C, Haas D (1993). Mobilization of chromosomes and nonconjugative plasmids by cointegrative mechanisms. In: DB Clewell (ed), Bacterial conjugation, pp 137–188. New York: Plenum Press.

    Google Scholar 

  12. Romero DA, Klaenhammer TR (1990). Characterization of insertion sequence IS946, an iso-ISSI element, isolated from the conjugative lactococcal plasmid pTR2030. J Bacteriol 172: 4151–4160.

    PubMed  CAS  Google Scholar 

  13. Romero DA, Klaenhammer TR (1993). Transposable elements in Lactococci: a review. J Dairy Sci 76: 1–19.

    Article  PubMed  CAS  Google Scholar 

  14. Romero DA, Slos P, Robert C, Castellino I, Mercenier A (1987). Conjugative mobilization as an alternative vector delivery system for lactic streptococci. Appl Environ Microbiol 53: 2405–2413.

    PubMed  CAS  Google Scholar 

  15. Sambrook J, Fritsch EF, Maniatis T (1989). Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  16. Sanders ME, Leonhard PJ, Sing WD, Klaenhammer TR (1986). Conjugal strategy for construction of fast acid-producing bacteriophage-resistant lactic streptococci for use in dairy fermentations. Appl Environ Microbiol 52: 1001–1007.

    PubMed  CAS  Google Scholar 

  17. Sing WD, Klaenhammer TR (1993). A strategy for rotation of different bacteriophage defenses in a lactococcal single-strain starter culture system. Appl Environ Microbiol 59: 365–372.

    PubMed  CAS  Google Scholar 

  18. Steele JL, McKay LL (1989). Conjugal transfer of genetic material in Lactococci: a review. J Dairy Sci 72: 3388–3397.

    Article  Google Scholar 

  19. Terzaghi BE, Sandine WE (1975). Improved medium for lactic streptococci and their bacteriophages. Appl Microbiol 29: 807–813.

    PubMed  CAS  Google Scholar 

  20. Walsh PM, McKay LL (1981). Recombinant plasmid associated with cell aggregation and high-frequency conjugation of Streptococcus lactis ML3. J Bacteriol 146: 937–944.

    PubMed  CAS  Google Scholar 

  21. Wilkins B, Lanka E (1993). DNA processing and replication during plasmid transfer between Gram-negative bacteria. In: D Clewell (ed), Bacterial conjugation, pp 105–129. New York: Plenum Press.

    Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Microbiology, University of Minnesota, Minneapolis, USA

    David A. Mills, Trevor G. Phister & Gary M. Dunny

  2. Department of Food Science and Nutrition, University of Minnesota, St Paul, Minnesota, USA

    Kathleen A. Baldwin

  3. Department of Food Science and Nutrition, University of Minnesota, 1334 Eckles Avenue, St Paul, MN, 55108, USA

    Larry L. McKay

Authors
  1. David A. Mills
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  2. Trevor G. Phister
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  3. Kathleen A. Baldwin
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  4. Gary M. Dunny
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  5. Larry L. McKay
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Corresponding author

Correspondence to Larry L. McKay .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Molecular Genetics, University of Vermont, Stafford Building, 05405, Burlington, Vermont, USA

    Paula M. Fives-Taylor

  2. Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana, USA

    Donald J. LeBlanc

  3. Department of Oral Biology, Indiana University School of Dentistry, Indianapolis, IN, USA

    Donald J. LeBlanc

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© 1998 Springer Science+Business Media Dordrecht

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Mills, D.A., Phister, T.G., Baldwin, K.A., Dunny, G.M., McKay, L.L. (1998). Characterization of the lactococcal conjugative element pRS01 using IS946-mediated mutagenesis. In: Fives-Taylor, P.M., LeBlanc, D.J. (eds) Methods for studying the genetics, molecular biology, physiology, and pathogenesis of the streptococci. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-2258-2_8

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  • DOI: https://doi.org/10.1007/978-94-017-2258-2_8

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-90-481-5262-9

  • Online ISBN: 978-94-017-2258-2

  • eBook Packages: Springer Book Archive

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