Skip to main content
SpringerLink
Log in

Conjugal transfer and expression of streptococcal transposons in Clostridium acetobutylicum

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

The transposon-containing streptococcal plasmids pAM211, pCF10, and pINY1275 have been transferred at high frequency (10-2–10-3 per recipient, selecting for tetracycline resistance) to the Gram-positive anaerobe Clostridium acetobutylicum. Selection in the presence of two antibiotics (tetracycline and erythromycin) with the plasmids pAM 180 and pINY1275 yielded only low numbers of transconjugants (≤10-8 per recipient). Matings were done by combining liquid and filter mating procedures under anaerobic conditions. No plasmid DNA could be detected in the transconjugants selected on a minimal medium in the presence of tetracycline. DNA-DNA hybridization experiments with restricted chromosomal DNA using biotinylated pAM120::Tn916 as probe revealed the presence of homologous sequences in the transconjugants but not in Clostridium acetobutylicum wild type. The transconjugants were used as donors in mating experiments with tetracycline-sensitive Bacillus subtilis and Streptococcus lactis subspec. diacetylactis. In both cases tetracycline-resistant strains were found. Transfer frequencies in these experiments were less than 10-7 per recipient.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Allcock ER, Reid SJ, Jones TD, Woods DR (1981) Autolytic activity and an autolysis-deficient mutant of Clostridum acetobutylicum. Appl Environ Microbiol 42:929–935

    Google Scholar 

  • Anderson DG, McKay LL (1983) Simple and rapid method for isolating large plasmid DNA from lactic streptococci. Appl Environ Microbiol 46:549–552

    Google Scholar 

  • Berg CM, Berg DE (1987) Uses of transposable elements and maps of known insertions. In: Neidhardt C, Ingraham JL, Low KB, Magasanik B, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella typhimurium. Cellular and molecular biology. American Society for Microbiology, Washington, DC, pp 1071–1109

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 7:1513–1523

    Google Scholar 

  • Bowring SN, Morris JG (1985) Mutagenesis of Clostridium acetobutylicum. J Appl Bacteriol 58:577–584

    Google Scholar 

  • Burdett, V, Inamine J, Rajagopalan S (1982) Heterogeneity of tetracycline resistance determinants in Streptococcus. J Bacteriol 149:995–1004

    Google Scholar 

  • Christie PJ, Korman RZ, Zahler SA, Adist JC, Dunny GM (1987) Two conjugation systems associated with Streptococcus faecalis plasmid pCF10: Identification of a conjugative transposon that transfers between S. faecalis and Bacillus subtilis. J Bacteriol 169:2529–2536

    Google Scholar 

  • Claus D, Lack P, Neu B (1983) DSM Catalogue of strains 1983, 3. edn. Gesellschaft für Biotechnologische Forschung mbH, Braunschweig

    Google Scholar 

  • Clewell DB, Gawron-Burke C (1986) Conjugative transposons and the dissemination of antibiotic resistance in streptococci. Ann Rev Microbiol 40:635–659

    Google Scholar 

  • Clewell DB, An FY, White BA, Gawron-Burke C (1985) Streptococcus faecalis sex pheromone (cAM 373) also produced by Staphylococcus aureus and identification of a conjugative transposon (Tn918). J Bacteriol 162:1212–1220

    Google Scholar 

  • Dürre P, Kuhn A, Gottschalk G (1986) Treatment with allyl alcohol selects specifically for mutants of Clostridium acetobutylicum defective in butanol synthesis. FEMS Microbiol Lett 36: 77–81

    Google Scholar 

  • Dybvig K, Cassell GH (1987) Transposition of Gram-positive transposon Tn916 in Acholeplasma laidlawii and Mycoplasma pulmonis. Science 235:1392–1394

    Google Scholar 

  • Gawron-Burke C, Clewell DB (1984) Regeneration of insertionally inactivated streptococcal DNA fragments after excision of transposon Tn916 in Escherichia coli: Strategy for targeting and cloning of genes from Gram-positive bacteria. J Bacteriol 159:214–221

    Google Scholar 

  • Hächler H, Kayser FH, Berger-Bächi B (1987) Homology of a transferable tetracycline resistance determinant of Clostridium difficile with Streptococcus (Enterococcus) faecalis transposon Tn916. Antimicrob Agents Chemother 31:1033–1038

    Google Scholar 

  • Hermann M, Fayolle F, Marchal R, Podvin L, Sebald M, Vandecasteele JP (1985) Isolation and characterization of butanol-resistant mutants of Clostridium acetobutylicum. Appl Environ Microbiol 50:1238–1243

    Google Scholar 

  • Hill C, Daly C, Fitzgerald GF (1985) Conjugative transfer of the transposon Tn919 to lactic acid bacteria. FEMS Microbiol Lett 30:115–119

    Google Scholar 

  • Hill C, Daly C, Fitzgerald GF (1987) Development of high-frequency delivery system for transposon Tn919 in lactic streptococci: random insertion in Streptococcus lactis subsp. diacetylactis 18–16. Appl Environ Microbiol 53:74–78

    Google Scholar 

  • Ivins BE, Welkos SL, Knudson GB, LeBlanc DJ (1988) Transposon Tn916 mutagenesis in Bacillus anthracis. Infect Immun 56:176–181

    Google Scholar 

  • Janati-Idrissi R, Junelles AM, El Kanouni A, Petitdemange H, Gay R (1987) Sélection de mutants de Clostridium acetobutylicum défectifs dans la production d'acétone. Ann Inst Pasteur/Microbiol 138:313–323

    Google Scholar 

  • Jones DT, Westhuizen A van der, Long S, Allcock ER Reid SJ, Woods DR, (1982) Solvent production and morphological changes in Clostridium acetobutylicum. Appl Environ Microbiol 43:1434–1439

    Google Scholar 

  • Jones DT, Jones WA, Woods DR (1985) Production of recombinants after protoplast fusion in Clostridium acetobytylicum P262. J Gen Microbiol 131:1213–1216

    Google Scholar 

  • Jones DT, Woods DR (1986a) Acetone-butanol fermentation revisited. Microbiol Rev 50:484–524

    Google Scholar 

  • Jones DT, Woods DR (1986b) Gene transfer, recombination and gene cloning in Clostridium acetobutylicum. Microbiol Sci 3:19–22

    Google Scholar 

  • Jones JM, Yost SC, Pattee PA (1987) Transfer of the conjugal tetracycline resistance transposon Tn916 from Streptococcus faecalis to Staphylococcus aureus and identification of some insertion sites in the staphylococcal chromosome. J Bacteriol 169:2121–2131

    Google Scholar 

  • Junelles AM, Janati-Idrissi R, El Kanouni A, Petitdemange H, Gay R (1987) Acetone-butanol fermentation by mutants selected for resistance to acetate and butyrate halogen analogues. Biotechnol Lett 9:175–178

    Google Scholar 

  • Largier ST, Long S, Santangelo JD, Jones DT, Woods DR (1985) Immobilized Clostridium acetobutylicum P262 mutants for solvent production. Appl Environ Microbiol 50:477–481

    Google Scholar 

  • Lee CK, Dürre, P, Hippe H, Gottschalk G (1987) Screening for plasmids in the genus Clostridium. Arch Microbiol 148: 107–114

    Google Scholar 

  • Lemmel SA (1985) Mutagenesis in Clostridium acetobutylicum. Biotechnol Lett 7:711–716

    Google Scholar 

  • Lin YL, Blaschek HP (1983) Butanol production by a butanol-tolerant strain of Clostridium acetobutylicum in extruded corn broth. Appl Environ Microbiol 45:966–973

    Google Scholar 

  • Lin YL, Blaschek HP (1984) Transformation of heat-treated Clostridium acetobutylicum protoplasts with pUB110 plasmid DNA. Appl Environ Microbiol 48:737–742

    Google Scholar 

  • Long S, Jones DT, Woods DR (1984) The relationship between sporulation and solvent production in Clostridium acetobutylicum P262. Biotechnol Lett 6:529–534

    Google Scholar 

  • Lovett PS, Keggins KM (1979) Bacillus subtilis as a host for molecular cloning. Meth Enzymol 68:342–357

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3:208–218

    Google Scholar 

  • Meinecke B, Bahl H, Gottschalk G (1984) Selection of an asporogenous strain of Clostridium acetobutylicum in continuous culture under phosphate limitation. Appl Environ Microbiol 48:1064–1065

    Google Scholar 

  • Neve H, Geis A, Teuber M (1987) Conjugation, a common plasmid transfer mechanism in lactic acid streptococci of dairy starter cultures. Syst Appl Microbiol 9:151–157

    Google Scholar 

  • O'Brien RW, Morris JG (1971a) Oxygen and the growth and metabolism of Clostridium acetobutylicum. J Gen Microbiol 68:307–318

    Google Scholar 

  • O'Brien RW, Morris JG (1971b) The ferredoxin-dependent reduction of chloramphenicol by Clostridium acetobutylicum. J Gen Microbiol 67:265–271

    Google Scholar 

  • Oultram JD, Young M (1985) Conjugal transfer of plasmid pAMβ1 from Streptococcus lactis and Bacillus subtilis to Clostridium acetobutylicum. FEMS Microbiol Lett 27:129–134

    Google Scholar 

  • Oultram JD, Davies A, Young M (1987) Conjugal transfer of a small plasmid from Bacillus subtilis to Clostridium acetobutylicum by cointegrate formation with plasmid pAMβ1. FEMS Microbiol Lett 42:113–119

    Google Scholar 

  • Reid SJ, Allcock ER, Jones DT, Woods DR (1983) Transformation of Clostridium acetobutylicum protoplasts with bacteriophage DNA. Appl Environ Microbiol 45:305–307

    Google Scholar 

  • Reysset G, Sebald M (1985) Conjugal transfer of plasmid antibiotic resistance from streptococci to Clostridium acetobutylicum. Ann Inst Pasteur/Microbiol 136B:275–282

    Google Scholar 

  • Roberts MC, Kenny GE (1987) Conjugal transfer of transposon Tn916 from Streptococcus faecalis to Mycoplasma hominis. J Bacteriol 169:3836–3839

    Google Scholar 

  • Rogers P (1984) Genetics and biochemistry of Clostridium relevant to development of fermentation processes. Adv Appl Microbiol 31:1–60

    Google Scholar 

  • Rogers P, Palosaari N (1987) Clostridium acetobutylicum mutants that produce butyraldehyde and altered quantities of solvents. Appl Environ Microbiol 53:2761–2766

    Google Scholar 

  • Senghas E, Jones JM, Yamamoto M, Gawron-Burke C, Clewell DB (1988) Genetic organization of the bacterial conjugative transposon Tn916. J Bacteriol 170:245–249

    Google Scholar 

  • Snedecor BR, Gomez RF (1983) Prospects for the use of genetic engineering to produce industrial chemicals with strict anaerobes. In: Wise DL (ed) Organic chemicals from biomass. Benjamin/Cummins Publ. Co., Inc., Menlo Park, pp 93–108

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Truffaut N, Sebald M (1983) Plasmid detection and isolation in strains of Clostridium acetobutylicum and related species. Mol Gen Genet 189:178–180

    Google Scholar 

  • Volk WA, Bizzini B, Jones KR, Macrina FL (1988) Inter- and intrageneric transfer of Tn916 between Streptococcus faecalis and Clostridium tetani. Plasmid 19:255–259

    Google Scholar 

  • Youngman PJ, Perkins JB, Losick R (1983) Genetic transposition and insertional mutagenesis in Bacillus subtilis with Streptococcus faecalis transposon Tn917. Proc Natl Acad Sci USA 80:2305–2309

    Google Scholar 

  • Yu PL, Pearce LE (1986) Conjugal transfer of streptococcal antibiotic resistance plasmids into Clostridium acetobutylicum. Biotechnol Lett 8:469–474

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Mikrobiologie, Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Germany

    Joachim Bertram & Peter Dürre

Authors
  1. Joachim Bertram
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter Dürre
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bertram, J., Dürre, P. Conjugal transfer and expression of streptococcal transposons in Clostridium acetobutylicum . Arch. Microbiol. 151, 551–557 (1989). https://doi.org/10.1007/BF00454874

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00454874

Key words

Search

Navigation