Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1983 Jan;153(1):200–210. doi: 10.1128/jb.153.1.200-210.1983

Genetic transformation of Streptococcus pneumoniae by DNA cloned into the single-stranded bacteriophage f1.

F Barany, J D Boeke
PMCID: PMC217358  PMID: 6571728

Abstract

A Staphylococcus aureus plasmid derivative, pFB9, coding for erythromycin and chloramphenicol resistance was cloned into the filamentous Escherichia coli phage f1. Recombinant phage-plasmid hybrids, designated plasmids, were isolated from E. coli and purified by transformation into Streptococcus pneumoniae. Single-stranded DNA was prepared from E. coli cells infected with two different plasmids, fBB101 and fBB103. Introduction of fully or partially single-stranded DNA into Streptococcus pneumoniae was studied, using a recipient strain containing an inducible resident plasmid. Such a strain could rescue the donor DNA marker. Under these marker rescue conditions, single-stranded fBB101 DNA gave a 1% transformation frequency, whereas the double-stranded form gave about a 31% frequency. Transformation of single-stranded fBB101 DNA was inhibited by competing double-stranded DNA and vice versa, indicating that single-stranded DNA interacts with the pneumococcus via the same binding site as used by double-stranded DNA. Heteroduplexed DNA containing the marker within a 70- or 800-base single-stranded region showed only slightly greater transforming activity than pure single-stranded DNA. In the absence of marker rescue, both strands of such imperfectly heteroduplexed DNA demonstrated transforming activity. Pure single-stranded DNA demonstrated low but significant transforming activity into a plasmid-free recipient pneumococcus.

Full text

PDF
200

Images in this article

202Image
on p.202
203Image
on p.203
207Image
on p.207

Selected References

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

  1. Barany F., Boeke J. D., Tomasz A. Staphylococcal plasmids that replicate and express erythromycin resistance in both Streptococcus pneumoniae and Escherichia coli. Proc Natl Acad Sci U S A. 1982 May;79(9):2991–2995. doi: 10.1073/pnas.79.9.2991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barany F., Tomasz A. Genetic transformation of Streptococcus pneumoniae by heterologous plasmid deoxyribonucleic acid. J Bacteriol. 1980 Nov;144(2):698–709. doi: 10.1128/jb.144.2.698-709.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Behnke D. Plasmid transformation of Streptococcus sanguis (Challis) occurs by circular and linear molecules. Mol Gen Genet. 1981;182(3):490–497. doi: 10.1007/BF00293940. [DOI] [PubMed] [Google Scholar]
  4. Boeke J. D. One and two codon insertion mutants of bacteriophage f1. Mol Gen Genet. 1981;181(3):288–291. doi: 10.1007/BF00425599. [DOI] [PubMed] [Google Scholar]
  5. Boeke J. D., Vovis G. F., Zinder N. D. Insertion mutant of bacteriophage f1 sensitive to EcoRI. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2699–2702. doi: 10.1073/pnas.76.6.2699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chaustova L. P., Grinius L. L., Griniuviene B. B., Jasaitis A. A., Kadziauskas J. P., Kiausinyte R. J. Studies on energy supply for genetic processes. Involvement of membrane potential in genetic transformation of Bacillus subtilis. Eur J Biochem. 1980 Jan;103(2):349–357. doi: 10.1111/j.1432-1033.1980.tb04321.x. [DOI] [PubMed] [Google Scholar]
  7. Gryczan T. J., Grandi G., Hahn J., Grandi R., Dubnau D. Conformational alteration of mRNA structure and the posttranscriptional regulation of erythromycin-induced drug resistance. Nucleic Acids Res. 1980 Dec 20;8(24):6081–6097. doi: 10.1093/nar/8.24.6081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gryczan T., Contente S., Dubnau D. Molecular cloning of heterologous chromosomal DNA by recombination between a plasmid vector and a homologous resident plasmid in Bacillus subtilis. Mol Gen Genet. 1980 Feb;177(3):459–467. doi: 10.1007/BF00271485. [DOI] [PubMed] [Google Scholar]
  9. Horinouchi S., Weisblum B. Nucleotide sequence and functional map of pC194, a plasmid that specifies inducible chloramphenicol resistance. J Bacteriol. 1982 May;150(2):815–825. doi: 10.1128/jb.150.2.815-825.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horinouchi S., Weisblum B. Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibodies. J Bacteriol. 1982 May;150(2):804–814. doi: 10.1128/jb.150.2.804-814.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Horinouchi S., Weisblum B. Posttranscriptional modification of mRNA conformation: mechanism that regulates erythromycin-induced resistance. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7079–7083. doi: 10.1073/pnas.77.12.7079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Horinouchi S., Weisblum B. The control region for erythromycin resistance: free energy changes related to induction and mutation to constitutive expression. Mol Gen Genet. 1981;182(2):341–348. doi: 10.1007/BF00269681. [DOI] [PubMed] [Google Scholar]
  13. Horiuchi K. Origin of DNA replication of bacteriophage f1 as the signal for termination. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5226–5229. doi: 10.1073/pnas.77.9.5226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Horiuchi K., Vovis G. F., Enea V., Zinder N. D. Cleavage map of bacteriophage f1: location of the Escherichia coli B-specific modification sites. J Mol Biol. 1975 Jun 25;95(2):147–165. doi: 10.1016/0022-2836(75)90388-5. [DOI] [PubMed] [Google Scholar]
  15. LACKS S. Molecular fate of DNA in genetic transformation of Pneumococcus. J Mol Biol. 1962 Jul;5:119–131. doi: 10.1016/s0022-2836(62)80067-9. [DOI] [PubMed] [Google Scholar]
  16. Lacks S., Greenberg B., Neuberger M. Identification of a deoxyribonuclease implicated in genetic transformation of Diplococcus pneumoniae. J Bacteriol. 1975 Jul;123(1):222–232. doi: 10.1128/jb.123.1.222-232.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lacks S., Greenberg B., Neuberger M. Role of a deoxyribonuclease in the genetic transformation of Diplococcus pneumoniae. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2305–2309. doi: 10.1073/pnas.71.6.2305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lai C. J., Weisblum B. Altered methylation of ribosomal RNA in an erythromycin-resistant strain of Staphylococcus aureus. Proc Natl Acad Sci U S A. 1971 Apr;68(4):856–860. doi: 10.1073/pnas.68.4.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lyons L. B., Zinder N. D. The genetic map of the filamentous bacteriophage f1. Virology. 1972 Jul;49(1):45–60. doi: 10.1016/s0042-6822(72)80006-0. [DOI] [PubMed] [Google Scholar]
  20. Macrina F. L., Jones K. R., Welch R. A. Transformation of Streptococcus sanguis with monomeric pVA736 plasmid deoxyribonucleic acid. J Bacteriol. 1981 May;146(2):826–830. doi: 10.1128/jb.146.2.826-830.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Miao R., Guild W. R. Competent Diplococcus pneumoniae accept both single- and double-stranded deoxyribonucleic acid. J Bacteriol. 1970 Feb;101(2):361–364. doi: 10.1128/jb.101.2.361-364.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Model P., Zinder N. D. In vitro synthesis of bacteriophage f1 proteins. J Mol Biol. 1974 Feb 25;83(2):231–251. doi: 10.1016/0022-2836(74)90389-1. [DOI] [PubMed] [Google Scholar]
  24. Morrison D. A., Guild W. R. Breakage prior to entry of donor DNA in Pneumococcus transformation. Biochim Biophys Acta. 1973 Apr 11;299(4):545–556. doi: 10.1016/0005-2787(73)90226-8. [DOI] [PubMed] [Google Scholar]
  25. Moses P. B., Boeke J. D., Horiuchi K., Zinder N. D. Restructuring the bacteriophage f1 genome: expression of gene VIII in the intergenic space. Virology. 1980 Jul 30;104(2):267–278. doi: 10.1016/0042-6822(80)90332-3. [DOI] [PubMed] [Google Scholar]
  26. Mottes M., Grandi G., Sgaramella V., Canosi U., Morelli G., Trautner T. A. Different specific activities of the monomeric and oligomeric forms of plasmid DNA in transformation of B. subtilis and E. coli. Mol Gen Genet. 1979 Jul 24;174(3):281–286. doi: 10.1007/BF00267800. [DOI] [PubMed] [Google Scholar]
  27. Murray N. E., Bruce S. A., Murray K. Molecular cloning of the DNA ligase gene from bacteriophage T4. II. Amplification and preparation of the gene product. J Mol Biol. 1979 Aug 15;132(3):493–505. doi: 10.1016/0022-2836(79)90271-7. [DOI] [PubMed] [Google Scholar]
  28. Novick R. P., Iordanescu S., Surdeanu M., Edelman I. Transduction-related cointegrate formation between Staphylococcal plasmids: a new type of site-specific recombination. Plasmid. 1981 Sep;6(2):159–172. doi: 10.1016/0147-619x(81)90064-0. [DOI] [PubMed] [Google Scholar]
  29. Novick R. P., Khan S. A., Murphy E., Iordanescu S., Edelman I., Krolewski J., Rush M. Hitchhiking transposons and other mobile genetic elements and site-specific recombination systems in Staphylococcus aureus. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):67–76. doi: 10.1101/sqb.1981.045.01.013. [DOI] [PubMed] [Google Scholar]
  30. Ohsumi M., Vovis G. F., Zinder N. D. The isolation and characterization of an in vivo recombinant between the filamentous bacteriophage f1 and the plasmid pSC101. Virology. 1978 Sep;89(2):438–449. doi: 10.1016/0042-6822(78)90186-1. [DOI] [PubMed] [Google Scholar]
  31. Saunders C. W., Guild W. R. Pathway of plasmid transformation in Pneumococcus: open circular and linear molecules are active. J Bacteriol. 1981 May;146(2):517–526. doi: 10.1128/jb.146.2.517-526.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Seto H., Tomasz A. Early stages in DNA binding and uptake during genetic transformation of pneumococci. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1493–1498. doi: 10.1073/pnas.71.4.1493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stassi D. L., Lopez P., Espinosa M., Lacks S. A. Cloning of chromosomal genes in Streptococcus pneumoniae. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7028–7032. doi: 10.1073/pnas.78.11.7028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tomasz A. Model for the mechanism controlling the expression of competent state in Pneumococcus cultures. J Bacteriol. 1966 Mar;91(3):1050–1061. doi: 10.1128/jb.91.3.1050-1061.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yamamoto K. R., Alberts B. M., Benzinger R., Lawhorne L., Treiber G. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology. 1970 Mar;40(3):734–744. doi: 10.1016/0042-6822(70)90218-7. [DOI] [PubMed] [Google Scholar]
  36. Zinder N. D., Boeke J. D. The filamentous phage (Ff) as vectors for recombinant DNA--a review. Gene. 1982 Jul-Aug;19(1):1–10. doi: 10.1016/0378-1119(82)90183-4. [DOI] [PubMed] [Google Scholar]

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

RESOURCES