Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1984 Feb;157(2):445–453. doi: 10.1128/jb.157.2.445-453.1984

Physical and genetic analyses of streptococcal plasmid pAM beta 1 and cloning of its replication region.

D J Leblanc, L N Lee
PMCID: PMC215268  PMID: 6319361

Abstract

Plasmid pAM beta 1, originally isolated from Streptococcus faecalis DS5, mediates resistance to the MLS (macrolide, lincosamide, and streptogramin B alpha) group of antibiotics. A restriction endonuclease map of the 26.5-kilobase (kb) pAM beta 1 molecule was constructed by using the enzymes AvaI, HpaII, EcoRI, PvuII, Kpn1, BstEII, HpaI, HhaI, and HindIII. A comparison of this map to those of four independently isolated deletion derivatives of pAM beta 1 located the MLS resistance determinant within a 2-kb DNA segment and at least one conjugative function within an 8-kb region. The 5.0-kb EcoRI-B fragment from pAM beta 1 was ligated onto the 4.0-kb Escherichia coli plasmid vector, pACKC1, and used to transform E. coli HB101. The 9.0-kb chimeric plasmid was then used to transform Streptococcus sanguis Challis with concurrent expression of the E. coli kanamycin resistance determinant. The 5.0-kb EcoRI-B fragment from pAM beta 1 was subsequently used as a vector to clone a streptomycin resistance determinant from a strain of Streptococcus mutans containing no detectable plasmid DNA. Subcloning experiments, using a HindIII partial digest of pAM beta 1 DNA, narrowed the replication region of this plasmid to a 2.95-kb fragment.

Full text

PDF
446

Images in this article

448Image
on p.448
449Image
on p.449
450Image
on p.450

Selected References

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

  1. Behnke D., Gilmore M. S. Location of antibiotic resistance determinants, copy control, and replication functions on the double-selective streptococcal cloning vector pGB301. Mol Gen Genet. 1981;184(1):115–120. doi: 10.1007/BF00271206. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  4. Burdett V., Inamine J., Rajagopalan S. Heterogeneity of tetracycline resistance determinants in Streptococcus. J Bacteriol. 1982 Mar;149(3):995–1004. doi: 10.1128/jb.149.3.995-1004.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chang A. C., Cohen S. N. Genome construction between bacterial species in vitro: replication and expression of Staphylococcus plasmid genes in Escherichia coli. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1030–1034. doi: 10.1073/pnas.71.4.1030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Clewell D. B., Yagi Y., Dunny G. M., Schultz S. K. Characterization of three plasmid deoxyribonucleic acid molecules in a strain of Streptococcus faecalis: identification of a plasmid determining erythromycin resistance. J Bacteriol. 1974 Jan;117(1):283–289. doi: 10.1128/jb.117.1.283-289.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Courvalin P., Fiandt M. Aminoglycoside-modifying enzymes of Staphylococcus aureus; expression in Escherichia coli. Gene. 1980 May;9(3-4):247–269. doi: 10.1016/0378-1119(90)90326-m. [DOI] [PubMed] [Google Scholar]
  10. El-Solh N., Bouanchaud D. H., Horodniceanu T., Roussel A., Chabbert Y. A. Molecular studies and possible relatedness between R plasmids from groups B and D streptococci. Antimicrob Agents Chemother. 1978 Jul;14(1):19–23. doi: 10.1128/aac.14.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Engel H. W., Soedirman N., Rost J. A., van Leeuwen W. J., van Embden J. D. Transferability of macrolide, lincomycin, and streptogramin resistances between group A, B, and D streptococci, Streptococcus pneumoniae, and Staphylococcus aureus. J Bacteriol. 1980 May;142(2):407–413. doi: 10.1128/jb.142.2.407-413.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gibson E. M., Chace N. M., London S. B., London J. Transfer of plasmid-mediated antibiotic resistance from streptococci to lactobacilli. J Bacteriol. 1979 Jan;137(1):614–619. doi: 10.1128/jb.137.1.614-619.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goldfarb D. S., Doi R. H., Rodriguez R. L. Expression of Tn9-derived chloramphenicol resistance in Bacillus subtilis. Nature. 1981 Sep 24;293(5830):309–311. doi: 10.1038/293309a0. [DOI] [PubMed] [Google Scholar]
  14. Hershfield V. Plasmids mediating multiple drug resistance in group B streptococcus: transferability and molecular properties. Plasmid. 1979 Jan;2(1):137–149. doi: 10.1016/0147-619x(79)90012-x. [DOI] [PubMed] [Google Scholar]
  15. Jagusztyn-Krynicka E. K., Smorawinska M., Curtiss R., 3rd Expression of Streptococcus mutans aspartate-semialdehyde dehydrogenase gene cloned into plasmid pBR322. J Gen Microbiol. 1982 May;128(5):1135–1145. doi: 10.1099/00221287-128-5-1135. [DOI] [PubMed] [Google Scholar]
  16. LeBlanc D. J., Hassell F. P. Transformation of Streptococcus sanguis Challis by plasmid deoxyribonucleic acid from Streptococcus faecalis. J Bacteriol. 1976 Oct;128(1):347–355. doi: 10.1128/jb.128.1.347-355.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. LeBlanc D. J., Hawley R. J., Lee L. N., St Martin E. J. "Conjugal" transfer of plasmid DNA among oral streptococci. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3484–3487. doi: 10.1073/pnas.75.7.3484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. LeBlanc D. J., Lee L. N. Characterization of two tetracycline resistance determinants in Streptococcus faecalis JH1. J Bacteriol. 1982 May;150(2):835–843. doi: 10.1128/jb.150.2.835-843.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Leblanc D. J., Cohen L., Jensen L. Transformation of group F streptococci by plasmid DNA. J Gen Microbiol. 1978 May;106(1):49–54. doi: 10.1099/00221287-106-1-49. [DOI] [PubMed] [Google Scholar]
  20. Leblanc D. J., Lee L. N. Rapid screening procedure for detection of plasmids in streptococci. J Bacteriol. 1979 Dec;140(3):1112–1115. doi: 10.1128/jb.140.3.1112-1115.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Macrina F. L., Jones K. R., Wood P. H. Chimeric streptococcal plasmids and their use as molecular cloning vehicles in Streptococcus sanguis (Challis). J Bacteriol. 1980 Sep;143(3):1425–1435. doi: 10.1128/jb.143.3.1425-1435.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Macrina F. L., Keeler C. L., Jr, Jones K. R., Wood P. H. Molecular characterization of unique deletion mutants of the streptococcal plasmid, pAM beta 1. Plasmid. 1980 Jul;4(1):8–16. doi: 10.1016/0147-619x(80)90079-7. [DOI] [PubMed] [Google Scholar]
  23. Malke H., Burman L. G., Holm S. E. Molecular cloning in streptococci: physical mapping of the vehicle plasmid pSM10 and demonstration of intergroup DNA transfer. Mol Gen Genet. 1981;181(2):259–267. doi: 10.1007/BF00268435. [DOI] [PubMed] [Google Scholar]
  24. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  25. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rubin E. M., Wilson G. A., Young F. E. Expression of thymidylate synthetase activity in Bacillus subtilis upon integration of a cloned gene from Escherichia coli. Gene. 1980 Aug;10(3):227–235. doi: 10.1016/0378-1119(80)90052-9. [DOI] [PubMed] [Google Scholar]
  27. Schottel J. L., Bibb M. J., Cohen S. N. Cloning and expression in streptomyces lividans of antibiotic resistance genes derived from Escherichia coli. J Bacteriol. 1981 Apr;146(1):360–368. doi: 10.1128/jb.146.1.360-368.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  29. Thayer R. E. An improved method for detecting foreign DNA in plasmids of Escherichia coli. Anal Biochem. 1979 Sep 15;98(1):60–63. doi: 10.1016/0003-2697(79)90705-x. [DOI] [PubMed] [Google Scholar]
  30. Vogelstein B., Gillespie D. Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci U S A. 1979 Feb;76(2):615–619. doi: 10.1073/pnas.76.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yag Y., Franke A. E., Clewell D. B. Plasmid-determined resistance to erythromycin: comparison of strains of streptococcus faecalis and streptococcus pyogenes with regard to plasmid hmology and resistance inducibility. Antimicrob Agents Chemother. 1975 Jun;7(6):871–873. doi: 10.1128/aac.7.6.871. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES