Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1981 Mar;31(3):1034–1043. doi: 10.1128/iai.31.3.1034-1043.1981

Characterization of the Streptococcus mutans plasmid pva318 cloned into Escherichia coli.

J B Hansen, Y Abiko, R Curtiss 3rd
PMCID: PMC351422  PMID: 6262244

Abstract

We further characterized the cryptic plasmid pVA318 of Streptococcus mutans. It had a contour length of 5.64 +/- 0.26 kilobases and a guanine-plus-cytosine content of 32 to 34 mol %. Upon cloning the pVA318 plasmid into the vector pBR322 in Escherichia coli, we made the following observations. The expression of tetracycline resistance by HindIII-cloned chimeras, where the insert was in the tetracycline resistance promotor, depended on the orientation of the pVA318 insert. Both HindIII-cloned chimeras segregated from polA(Ts) cells at a nonpermissive temperature. Chimeric molecules cloned with PstI initially showed much instability; the reason for this is unknown, although stable variants were obtained. Both HindIII-cloned variants and a PstI-cloned chimera produced a pVA318-specific protein of approximately 20,000 molecular weight in E. coli minicells. The biological function of this protein is not known; it had no bacteriocin activity against S. mutans or group A Streptococcus indicator strains, and it did not appear in the E. coli periplasm. We constructed a map of pVA318 for restriction endonucleases HindIII, HpaI, PstI, and HaeIII. A previously reported BamHI site in pVA318 did not appear in the pVA318 portion of any of our chimeric clones.

Full text

PDF
1034

Images in this article

1039Image
on p.1039
1040Image
on p.1040
1040Image
on p.1040

Selected References

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

  1. Aaij C., Borst P. The gel electrophoresis of DNA. Biochim Biophys Acta. 1972 May 10;269(2):192–200. doi: 10.1016/0005-2787(72)90426-1. [DOI] [PubMed] [Google Scholar]
  2. Behnke D., Ferretti J. J. Molecular cloning of an erythromycin resistance determinant in streptococci. J Bacteriol. 1980 Nov;144(2):806–813. doi: 10.1128/jb.144.2.806-813.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  5. Dunny G. M., Birch N., Hascall G., Clewell D. B. Isolation and characterization of plasmid deoxyribonucleic acid from Streptococcus mutans. J Bacteriol. 1973 Jun;114(3):1362–1364. doi: 10.1128/jb.114.3.1362-1364.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Eckhardt T. A rapid method for the identification of plasmid desoxyribonucleic acid in bacteria. Plasmid. 1978 Sep;1(4):584–588. doi: 10.1016/0147-619x(78)90016-1. [DOI] [PubMed] [Google Scholar]
  7. Fling M. E., Elwell L. P. Protein expression in Escherichia coli minicells containing recombinant plasmids specifying trimethoprim-resistant dihydrofolate reductases. J Bacteriol. 1980 Feb;141(2):779–785. doi: 10.1128/jb.141.2.779-785.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frazer A. C., Curtiss R., 3rd Production, properties and utility of bacterial minicells. Curr Top Microbiol Immunol. 1975;69:1–84. doi: 10.1007/978-3-642-50112-8_1. [DOI] [PubMed] [Google Scholar]
  9. Freifelder D., Dewitt R. Thermal hydrolysis as a means of opening supercoiled circles of DNA. Gene. 1977 Jul;1(5-6):385–387. doi: 10.1016/0378-1119(77)90042-7. [DOI] [PubMed] [Google Scholar]
  10. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guerry P., LeBlanc D. J., Falkow S. General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol. 1973 Nov;116(2):1064–1066. doi: 10.1128/jb.116.2.1064-1066.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hansen J. B., Olsen R. H. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J Bacteriol. 1978 Jul;135(1):227–238. doi: 10.1128/jb.135.1.227-238.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hazelbauer G. L., Harayama S. Mutants in transmission of chemotactic signals from two independent receptors of E. coli. Cell. 1979 Mar;16(3):617–625. doi: 10.1016/0092-8674(79)90035-7. [DOI] [PubMed] [Google Scholar]
  14. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Levy S. B. Very stable prokaryotic messenger RNA in chromosomeless Escherichia coli minicells. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2900–2904. doi: 10.1073/pnas.72.8.2900. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Macrina F. L., Kopecko D. J., Jones K. R., Ayers D. J., McCowen S. M. A multiple plasmid-containing Escherichia coli strain: convenient source of size reference plasmid molecules. Plasmid. 1978 Jun;1(3):417–420. doi: 10.1016/0147-619x(78)90056-2. [DOI] [PubMed] [Google Scholar]
  20. Macrina F. L., Reider J. L., Virgili S. S., Kopecko D. J. Survey of the extrachromosomal gene pool of Streptococcus mutans. Infect Immun. 1977 Jul;17(1):215–226. doi: 10.1128/iai.17.1.215-226.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Macrina F. L., Scott C. L. Evidence for a disseminated plasmid in Streptococcus mutans. Infect Immun. 1978 Apr;20(1):296–302. doi: 10.1128/iai.20.1.296-302.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Macrina F. L., Virgili S. S., Scott C. L. Extrachromosomal gene systems in Streptococcus mutans. Adv Exp Med Biol. 1978;107:859–868. doi: 10.1007/978-1-4684-3369-2_96. [DOI] [PubMed] [Google Scholar]
  23. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nelson T., Brutlag D. Addition of homopolymers to the 3'-ends of duplex DNA with terminal transferase. Methods Enzymol. 1979;68:41–50. doi: 10.1016/0076-6879(79)68005-9. [DOI] [PubMed] [Google Scholar]
  25. Ohtsubo H., Ohtsubo E. Nucleotide sequence of an insertion element, IS1. Proc Natl Acad Sci U S A. 1978 Feb;75(2):615–619. doi: 10.1073/pnas.75.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  27. Smith D. I., Blattner F. R., Davies J. The isolation and partial characterization of a new restriction endonuclease from Providencia stuartii. Nucleic Acids Res. 1976 Feb;3(2):343–353. doi: 10.1093/nar/3.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  29. VINOGRAD J., HEARST J. E. Equilibrium sedimentation of macromolecules and viruses in a density gradient. Fortschr Chem Org Naturst. 1962;20:373–422. [PubMed] [Google Scholar]
  30. Woodward R. S., Lebowitz J. A revised equation relating DNA buoyant density to guanine plus cytosine content. J Biochem Biophys Methods. 1980 May;2(5):307–309. doi: 10.1016/0165-022x(80)90055-x. [DOI] [PubMed] [Google Scholar]
  31. Zasloff M., Ginder G. D., Felsenfeld G. A new method for the purification and identification of covalently closed circular DNA molcules. Nucleic Acids Res. 1978 Apr;5(4):1139–1152. doi: 10.1093/nar/5.4.1139. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES