1887

Abstract

An excision reporter plasmid was constructed to characterize the intracellular mobility of Tn in various Gram-positive bacteria. The reporter component of this plasmid is a chloramphenicol-resistance gene which has been insertionally inactivated with the integrative vector pAT112 containing the attachment site of Tn Tn-mediated excision of pAT112, to produce clones resistant to chloramphenicol, was detected in BM4110, L028-Str and BM120, but not in MG1363-RF or in BM124, and always depended upon the ability of the bacterial host to generate circular forms of the transposon. The results suggest that (i) the excision event, although required, is not sufficient for conjugal transfer to occur and (ii) there is no linear relationship between the donor potential of strains and either the excision frequency of pAT112 or the copy number of Tn circular intermediates per cell in these hosts. Excision of pAT112 occurred mainly during the late exponential phase of growth of and monocytogenes and this recombination event was not stimulated by heat shock, salt and alcohol stresses or by the presence of tetracycline in the medium.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-4-1253
1997-04-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/4/mic-143-4-1253.html?itemId=/content/journal/micro/10.1099/00221287-143-4-1253&mimeType=html&fmt=ahah

References

  1. Bringel, F, Van Alstine G. L., Scott J. R. 1991; A host factor absent from Lactococcus lactis subspecies lactis MG1363 is required for conjugative transposition.. Mol Microbiol 5:2983–2993
    [Google Scholar]
  2. buu-Hoi A. Y., Horodniceanu T. 1980; Conjugative transfer of multiple antibiotic resistance markers in Streptococcus pneumoniae . J Bacteriol 143:313–320
    [Google Scholar]
  3. Churchward G.,, Belin D., Nagamine Y. 1984; A pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectors.. Gene 31:165–171
    [Google Scholar]
  4. Clermont D., Horaud T. 1994; Genetic and molecular studies of a composite chromosomal element (Tn3705) containing a Tn92 6-modified structure (Tn3704) in Streptococcus anginosus F22.. Plasmid 31:40–48
    [Google Scholar]
  5. Clewell D. B.,, Flannagan S. E., Jaworski D. D. 1995; Unconstrained bacterial promiscuity : the Tn916-Tn2545 family of conjugative transposons.. Trends Microbiol 3:229–236
    [Google Scholar]
  6. Courvalin, P., Carlier C. 1986; Transposable multiple antibiotic resistance in Streptococcus pneumoniae.. Mol Gen Genet 205:291–297
    [Google Scholar]
  7. Courvalin, P., Carlier C. 1987; Tn1545: a conjugative shuttle transposon.. Mol Gen Genet 206:259–264
    [Google Scholar]
  8. David, F., de Cespedes G., Delbos F., Horaud, T. 1993; Diversity of chromosomal genetic elements and gene identification in antibiotic-resistant strains of Streptococcus pneumoniae and Streptococcus bovis.. Plasmid 29:147–153
    [Google Scholar]
  9. Doucet-Populaire F.,, Trieu-Cuot P., Dosbaa I., Andremont A., Courvalin P. 1991; Inducible transfer of conjugative transposon Tn1545 from Enterococcus faecalis to Listeria monocytogenes in the digestive tract of gnotobiotic mice.. Antimicrob Agents Chemothe 35:185–187
    [Google Scholar]
  10. Flannagan S., Clewell, D. B. 1991; Conjugative transfer of Tn916 in Enterococcus faecalis : trans activation of homologous transposons. J Bacteriol 173:7136–7141
    [Google Scholar]
  11. Flannagan S.E, Zitzow, L. A.,Su Y. A., Clewell, D. B. 1994; Nucleotide sequence of the 18-kb conjugative transposon Tn916 from Enterococcus faecalis.. Plasmid 32:350–354
    [Google Scholar]
  12. Jacobs A.E, Hobbs S.J., Clewell, D. B. 1974; Conjugal transfer of plasmidborne multiple antibiotic resistance in Streptococcus faecalis var. zymogenes.. J Bacteriol 117:360–372
    [Google Scholar]
  13. Jaworski D. D., Clewell D. B. 1994; Evidence that the coupling sequences play a frequency-determining role in conjugative transposition of Tn916 in Enterococcus faecalis.. J Bacteriol 176:3328–3335
    [Google Scholar]
  14. Jaworski D. D., Clewell D. B. 1995; A functional origin of transfer (oriT) on the conjugative transposon Tn916. J Bacteriol 177:6644–6651
    [Google Scholar]
  15. Langella P.,, Le Loir Y.,, Ehrlich S. D., Gruss A. 1993; Efficient plasmid mobilization by pIP501 in Lactococcus lactis subsp. lactis. . J Bacteriol 175:5806–5813
    [Google Scholar]
  16. Le Bouguenec C.,, Horaud T.,, Bieth, G., Colimon R., Dauguet C. 1984; Translocation of antibiotic resistance markers of a plasmid-free Streptococcus pyogenes (group A) strain into different streptococcal hemolysin plasmids.. Mol Gen Genet 194:377–387
    [Google Scholar]
  17. Le Bouguenec .,, de Cespedes G.,, Horaud, T.,. 1990; Presence of chromosomal elements resembling the composite structure Tn3701 in streptococci.. J Bacteriol 172:727–734
    [Google Scholar]
  18. Manganelli, R.,, Romano L.,, Ricci, S.,, Zazzi, M., Pozzi, G. 1995; Dosage of Tn916 circular intermediates in Enterococcus faecalis. . Plasmid 34:48–57
    [Google Scholar]
  19. Poyart C., Trieu-Cuot, P. 1994; Heterogeneric conjugal transfer of the pheromone-responsive plasmid pIP964 (IncHlyI) of Enterococcus faecalis in the apparent absence of pheromone induction.. FEMS Microbiol Lett 122:173–180
    [Google Scholar]
  20. Poyart C., Celli, J., Trieu-Cuot P. 1995; Conjugative transposition of Tn916-related elements from Enterococcus faecalis to Escherichia coli and Pseudomonas fluorescens.. Antimicrob Agents Chemother 39:500–506
    [Google Scholar]
  21. Poyart-Salmeron C., Trieu-Cuot P., Carlier C., Courvalin P. 1989; Molecular characterization of two proteins involved in the excision of the conjugative transposon Tn1545 : homologies with other site-specific recombinases.. EMBO J 8:2425–2433
    [Google Scholar]
  22. Poyart-Salmeron C., Trieu-Cuot P., Carlier C., Courvalin P. 1990; The integration-excision system of the streptococcal transposon Tn1545 is structurally and functionally related to those of lambdoid phages. Mol Microbiol 4:1513–1521
    [Google Scholar]
  23. Provvedi R.,, Manganelli R., Pozzi G. 1996; Characterization of conjugative transposon Tn5251 of Streptococcus pneumoniae.. FEMS Microbiol Lett 135:231–236
    [Google Scholar]
  24. Rice L. B.,, Marshall, S. H., Carias L. L. 1992; Tn5381, a conjugative transposon identifiable as a circular form in Enterococcus faecalis.. J Bacteriol 174:7308–7315
    [Google Scholar]
  25. Rice L. B.,, Carias L. L., Marshall, S. H. 1995; Tn5384, a composite enterococcal mobile element conferring resistance to erythromycin and gentamicin whose ends are directly repeated copies of IS256.. Antimicrob Agents Chemother 39:1147–1153
    [Google Scholar]
  26. Sambrook J.,, Fritsch E. F., Maniatis, T. 1989; Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
    [Google Scholar]
  27. Scott J. R., Churchward, G. G. 1995; Conjugative transposition.. Annu Rev Microbiol 49:367–397
    [Google Scholar]
  28. Showsh S. A., Andrew, R. E. J. 1992; Tetracycline enhances Tn916-mediated conjugal transfer.. Plasmid 28:213–224
    [Google Scholar]
  29. Simon R.,, Priefer U., Puhler A. 1983; A broad host range mobilization system for in vivo genetic engineering : transposon mutagenesis in Gram-negative bacteria.. Biotechnology 1:784–794
    [Google Scholar]
  30. Storrs M. J., Poyart-Salmeron C.,, Trieu-Cuot P., Courvalin P. 1991; Conjugative transposition of Tn916 requires the excisive and integrative activities of the transposon-encoded integrase Int- Tn.. J Bacteriol 173:4347–4352
    [Google Scholar]
  31. Torres O. R.,, Korman R. Z.,, Zahler S. A., Dunny G. M. 1991; The conjugative transposon Tn925 : enhancement of conjugal transfer by tetracycline in Enterococcus faecalis and mobilization of chromosomal genes in Bacillus subtilis and E. faecalis. . Mol Gen Genet 225:395–400
    [Google Scholar]
  32. Trieu-Cuot, P.,, Carlier, C., Courvalin, P. 1988; Conjugative plasmid transfer from Enterococcus faecalis to Escherichia coli. . J Bacteriol 170:4388–4391
    [Google Scholar]
  33. Trieu-Cuot, P.,, Carlier, C., Poyart-Salmeron C., Courvalin, P. 1991; An integrative vector exploiting the transposition properties of Tn1545 for insertional mutagenesis and cloning of genes from Gram-positive bacteria.. Gene 106:21–27
    [Google Scholar]
  34. Trieu-Cuot, P.,, de Cespedes G., Horaud, T. 1992; Nucleotide sequence of the chloramphenicol resistance determinant of the streptococcal plasmid pIP501.. Plasmid 28:272–276
    [Google Scholar]
  35. Zhang, H.,, Scholl, R., Browse, J., Somerville, C. 1988; Doublestranded DNA sequencing as a choice for DNA sequencing.. Nucleic Acids Res 16:120
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-4-1253
Loading
/content/journal/micro/10.1099/00221287-143-4-1253
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error