Skip to main content
Log in

Isolation and purification of plasmids from Bacteriodes fragilis using rubidium trichloroacetate density gradient centrifugation

  • Short Communication
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

A rapid and easy final purification method is described for the isolation of plasmids from B. fragilis. Using RbTCA density gradient centrifugation in an airfuge ultracentrifuge ccc plasmid DNA can be separated from RNA, residual chromosomal DNA, linear and oc plasmid DNA. Pure ccc plasmid DNA is obtained from cultures of between 1 ml and 2 l in less than one day.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

References

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    Google Scholar 

  • Burke RL, Bauer WR (1977) The properies of native and denatured DNA in buoyant rubidium trichloroacetate at neutral pH. Nucleic Acids Res 4:1891–1909

    Google Scholar 

  • Burke RL, Bauer WR (1978) The helix-coil transition of closed and nicked DNAs in aqueous neutral trichloroacetate solutions. Nucleic Acids Res 5:4819–4836

    Google Scholar 

  • Burke RL, Bauer WR (1980) The early melting of closed duplex DNA: analysis by banding in buoyant neutral rubidium trichloroacetate. Nucleic Acids Res 8:1145–1165

    Google Scholar 

  • Clewell DB, Helinski DR (1969) Supercoiled circular DNA-protein complex in Escherichia coli: Purification and induced conversion to an open circular DNA form. Biochemistry 62:1157–1166

    Google Scholar 

  • Colman A, Byers MJ, Primose SB, Lyons A (1978) Rapid purification of plasmid DNAs by hydroxyapatite chromatography. Eur J Biochem 91:303–310

    Google Scholar 

  • Cornelis P, Digneffe C, Willemot K, Colson C (1981) Purification of Escherichia coli amplificable plasmids by highsalt sepharose chromatography. Plasmid 5:221–223

    Google Scholar 

  • Guerry P, LeBlanc DJ, Falkow S (1973) General method for the isolation of plasmid deoxyribonucleic acid. J. Bacteriol 116:1064–1066

    Google Scholar 

  • Guiney DG, Davies CE (1975) Isolation of plasmid deoxyribonucleic acid from two strains of bacteroides. J Bacteriol 124:503–510

    Google Scholar 

  • Guiney DG, Davies CE (1978) Identification of a conjugative R plasmid in Bacteroides ochraceus capable of transfer to Escherichia coli. Nature 274:181–182

    Google Scholar 

  • Hansen JB, Olsen RH (1978) Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMGl and pMG5. J Bacteriol 135:227–238

    Google Scholar 

  • Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197

    Google Scholar 

  • Kado CI, Liu ST (1981) Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 145:1365–1373

    Google Scholar 

  • Klein RD, Selsing E, Wells RD (1980) A rapid microscale technique for isolation of recombinant plasmid DNA suitable for restriction enzyme analysis. Plasmid 3:88–91

    Google Scholar 

  • Mays TD, Smith CJ, Welch RA, Delfini C, Macrina L (1982) Novel antibiotic resistance transfer in Bacteroides. Antimicrob Agents Chemother 21:110–118

    Google Scholar 

  • Meyers JA, Sanchez D, Elwell LP, Falkow S (1976) Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J Bacteriol 127:1529–1537

    Google Scholar 

  • Pakroppa W, Goebel W, Müller W (1975) Isolation of closedcircular duplex DNA by chromatography on hydroxyapatite in the presence of ethidium bromide. Anal Biochem 67:372–383

    Google Scholar 

  • Palchaudhuri S, Bawdon RE (1980) Plasmid mediated clindamycin resistance in Bacterioides fragilis. In: Stuttgard C, Rozee KR (eds) Plasmids and transposons. Academic Press, New York, London, pp 129–136

    Google Scholar 

  • Privitera G, Botta G, Sebald M (1981) Macrolide, lincosamide, streptogramin and tetracycline transferable resistance in Bacteroides fragilis group. J Antimicrob Chemother 8:Suppl D 87–94

    Google Scholar 

  • Ranhand FM (1982) A simple method for the qualitative demonstration of large and small plasmid deoxyribonuceic acids in gram-positive and gram-negative bacteria. Anal Biochem 122:426–432

    Google Scholar 

  • Rashtchian A, Dubes GR, Booth SJ (1982) Tetracycline-inducible transfer of tetracycline resistance in Bacteroides fragilis in the absence of detectable plasmid DNA. J Bacteriol 150:141–147

    Google Scholar 

  • Rashtchian A, Booth SJ (1981) Stability in Escherichia coli of an antibiotic resistance plasmid from Bacteroides fragilis. J Bacteriol 146:121–127

    Google Scholar 

  • Rotimi VO, Duerden BI, Hafiz S (1981) Transferable plasmidmediated antibiotic resistance in Bacteroides. J Med Microbiol 14:359–370

    Google Scholar 

  • Sharp PA, Hsu M-T, Ohtsubo E, Davidson N (1972) Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. J Mol Biol 71:471–497

    Google Scholar 

  • Stiffler PW, Keller R, Traub T (1974) Isolation and characterisation of several cryptic plasmids from clinical isolates of Bacteroids fragilis. J Infect Dis 130:544–548

    Google Scholar 

  • Stougaard P, Molin S (1981) Vertical dye-buoyant density gradients for rapid analysis and preparation of plasmid DNA. Anal Biochem 118:191–193

    Google Scholar 

  • Talley FP, Snydman DR, Gorbach S, Malamy M (1979) Plasmid mediated, transferable resistance to clindamycin and erythromycin in Bacteroides fragilis. J Infect Dis 139:83–88

    Google Scholar 

  • Tinnel WH, Macrina FL (1976) Extrachromosomal elements in a variety of strains representing the Bacteroides fragilis group of organisms. Infect Immun 14:955–964

    Google Scholar 

  • Wallace BL, Bradley JE, Rogolsky M (1981) Plasmid analyses in clinical isolates of Bacteroides fragilis and other Bacterioides species. J Clin Microbiol 14:383–388

    Google Scholar 

  • Welch R, Jones K, Macrina FL (1979) Transferable lincosamidemacrolide resistance in Bacteroides. Plasmid 2:261–268

    Google Scholar 

  • Welch RA, Macrina FL (1981) Physical characterisation of Bacteroides fragilis R plasmid pBF4, J Bacteriol 145:867–872

    Google Scholar 

  • Welch RA, Macrina FL (1980) Deletion analysis of a transferable Bacteroides R plasmid. In: Shuttard C, Rozee KR (eds) Plasmids and transposons. Academic Press, New-York, London, pp 137–144

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by E. Bautz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zöllner, E.J., Eichel-Streiber, C.v. & Schwindling, FP. Isolation and purification of plasmids from Bacteriodes fragilis using rubidium trichloroacetate density gradient centrifugation. Molec Gen Genet 189, 169–171 (1983). https://doi.org/10.1007/BF00326072

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00326072

Keywords

Navigation