Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1986 Jun;166(3):972–977. doi: 10.1128/jb.166.3.972-977.1986

Cloning and physical characterization of chromosomal conjugative elements in streptococci.

M N Vijayakumar, S D Priebe, G Pozzi, J M Hageman, W R Guild
PMCID: PMC215220  PMID: 3011756

Abstract

We used a directed insertion method to introduce a nonreplicating vector plasmid into the large conjugative cat-tet element found in the chromosome of Streptococcus pneumoniae BM6001 and derivatives. To direct insertion preferentially to the conjugative element, we transferred it by conjugation to Streptococcus faecalis and then used DNA from this strain as a source of restriction nuclease fragments for ligation to digests of the vector pVA891, which can replicate in Escherichia coli but not in streptococci. This ligation mix was used to transform pneumococcal cells carrying the cat-tet element, with selection for the erythromycin resistance carried by pVA891. Eight such isolates were found, and transformation and conjugation tests showed that in each case the vector had inserted into the conjugative element, as expected. DNA from these pneumococcal strains generated a variety of E. coli plasmids which provide tools for obtaining a detailed restriction map and for defining other structural features of the streptococcal conjugative element.

Full text

PDF
976

Selected References

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

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clewell D. B. Plasmids, drug resistance, and gene transfer in the genus Streptococcus. Microbiol Rev. 1981 Sep;45(3):409–436. doi: 10.1128/mr.45.3.409-436.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Franke A. E., Clewell D. B. Evidence for a chromosome-borne resistance transposon (Tn916) in Streptococcus faecalis that is capable of "conjugal" transfer in the absence of a conjugative plasmid. J Bacteriol. 1981 Jan;145(1):494–502. doi: 10.1128/jb.145.1.494-502.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gawron-Burke C., Clewell D. B. A transposon in Streptococcus faecalis with fertility properties. Nature. 1982 Nov 18;300(5889):281–284. doi: 10.1038/300281a0. [DOI] [PubMed] [Google Scholar]
  5. Guild W. R., Shoemaker N. B. Mismatch correction in pneumococcal transformation: donor length and hex-dependent marker efficiency. J Bacteriol. 1976 Jan;125(1):125–135. doi: 10.1128/jb.125.1.125-135.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  7. Horodniceanu T., Bougueleret L., Bieth G. Conjugative transfer of multiple-antibiotic resistance markers in beta-hemolytic group A, B, F, and G streptococci in the absence of extrachromosomal deoxyribonucleic acid. Plasmid. 1981 Mar;5(2):127–137. doi: 10.1016/0147-619x(81)90014-7. [DOI] [PubMed] [Google Scholar]
  8. Inamine J. M., Burdett V. Structural organization of a 67-kilobase streptococcal conjugative element mediating multiple antibiotic resistance. J Bacteriol. 1985 Feb;161(2):620–626. doi: 10.1128/jb.161.2.620-626.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Macrina F. L., Evans R. P., Tobian J. A., Hartley D. L., Clewell D. B., Jones K. R. Novel shuttle plasmid vehicles for Escherichia-Streptococcus transgeneric cloning. Gene. 1983 Nov;25(1):145–150. doi: 10.1016/0378-1119(83)90176-2. [DOI] [PubMed] [Google Scholar]
  10. Morrison D. A., Trombe M. C., Hayden M. K., Waszak G. A., Chen J. D. Isolation of transformation-deficient Streptococcus pneumoniae mutants defective in control of competence, using insertion-duplication mutagenesis with the erythromycin resistance determinant of pAM beta 1. J Bacteriol. 1984 Sep;159(3):870–876. doi: 10.1128/jb.159.3.870-876.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Méjean V., Claverys J. P., Vasseghi H., Sicard A. M. Rapid cloning of specific DNA fragments of Streptococcus pneumoniae by vector integration into the chromosome followed by endonucleolytic excision. Gene. 1981 Nov;15(2-3):289–293. doi: 10.1016/0378-1119(81)90139-6. [DOI] [PubMed] [Google Scholar]
  12. Pozzi G., Guild W. R. Modes of integration of heterologous plasmid DNA into the chromosome of Streptococcus pneumoniae. J Bacteriol. 1985 Mar;161(3):909–912. doi: 10.1128/jb.161.3.909-912.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sanger F., Air G. M., Barrell B. G., Brown N. L., Coulson A. R., Fiddes C. A., Hutchison C. A., Slocombe P. M., Smith M. Nucleotide sequence of bacteriophage phi X174 DNA. Nature. 1977 Feb 24;265(5596):687–695. doi: 10.1038/265687a0. [DOI] [PubMed] [Google Scholar]
  14. Sanger F., Coulson A. R., Hong G. F., Hill D. F., Petersen G. B. Nucleotide sequence of bacteriophage lambda DNA. J Mol Biol. 1982 Dec 25;162(4):729–773. doi: 10.1016/0022-2836(82)90546-0. [DOI] [PubMed] [Google Scholar]
  15. Shoemaker N. B., Guild W. R. Destruction of low efficiency markers is a slow process occurring at a heteroduplex stage of transformation. Mol Gen Genet. 1974;128(4):283–290. doi: 10.1007/BF00268516. [DOI] [PubMed] [Google Scholar]
  16. Shoemaker N. B., Smith M. D., Guild W. R. DNase-resistant transfer of chromosomal cat and tet insertions by filter mating in Pneumococcus. Plasmid. 1980 Jan;3(1):80–87. doi: 10.1016/s0147-619x(80)90036-0. [DOI] [PubMed] [Google Scholar]
  17. Shoemaker N. B., Smith M. D., Guild W. R. Organization and transfer of heterologous chloramphenicol and tetracycline resistance genes in pneumococcus. J Bacteriol. 1979 Aug;139(2):432–441. doi: 10.1128/jb.139.2.432-441.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith M. D., Guild W. R. Improved method for conjugative transfer by filter mating of Streptococcus pneumoniae. J Bacteriol. 1980 Oct;144(1):457–459. doi: 10.1128/jb.144.1.457-459.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Smith M. D., Hazum S., Guild W. R. Homology among tet determinants in conjugative elements of streptococci. J Bacteriol. 1981 Oct;148(1):232–240. doi: 10.1128/jb.148.1.232-240.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vijayakumar M. N., Priebe S. D., Guild W. R. Structure of a conjugative element in Streptococcus pneumoniae. J Bacteriol. 1986 Jun;166(3):978–984. doi: 10.1128/jb.166.3.978-984.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES