Characterization of the Streptococcus mutans plasmid pVA318 cloned into Escherichia coli

J. B. Hansen, Y. Abiko, R. Curtiss

Research output: Contribution to journalArticle

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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 promoter, 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.

Original languageEnglish (US)
Pages (from-to)1034-1043
Number of pages10
JournalInfection and Immunity
Volume31
Issue number3
StatePublished - 1981
Externally publishedYes

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Streptococcus mutans
Tetracycline Resistance
Plasmids
Escherichia coli
Deoxyribonuclease HindIII
Periplasm
Bacteriocins
DNA Restriction Enzymes
Base Composition
Streptococcus
Organism Cloning
Proteins
Clone Cells
Molecular Weight
Temperature

ASJC Scopus subject areas

  • Immunology

Cite this

Characterization of the Streptococcus mutans plasmid pVA318 cloned into Escherichia coli. / Hansen, J. B.; Abiko, Y.; Curtiss, R.

In: Infection and Immunity, Vol. 31, No. 3, 1981, p. 1034-1043.

Research output: Contribution to journalArticle

Hansen, JB, Abiko, Y & Curtiss, R 1981, 'Characterization of the Streptococcus mutans plasmid pVA318 cloned into Escherichia coli', Infection and Immunity, vol. 31, no. 3, pp. 1034-1043.
Hansen, J. B. ; Abiko, Y. ; Curtiss, R. / Characterization of the Streptococcus mutans plasmid pVA318 cloned into Escherichia coli. In: Infection and Immunity. 1981 ; Vol. 31, No. 3. pp. 1034-1043.
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