1887

Abstract

Summary: Rapid methods for the analysis of degradative plasmids in are described. Bacterial lysates prepared with alkaline sodium dodecyl sulphate were vigorously stirred in the presence of antifoam agent. Samples were subjected to agarose gel electrophoresis to detect plasmid DNA. After a short period of centrifugation on a sucrose gradient, the lysates yielded plasmid DNA of sufficient purity for restriction endonuclease digestion without further treatment. The methods have proved most useful in the extraction of previously undetected plasmid DNA, some of which appears to be unstable.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-124-2-433
1981-06-01
2021-05-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/124/2/mic-124-2-433.html?itemId=/content/journal/micro/10.1099/00221287-124-2-433&mimeType=html&fmt=ahah

References

  1. Barnes W.M. 1977; Plasmid detection and sizing in single colony lysates. Science 195:393–394
    [Google Scholar]
  2. Baxter-Gabbard K.L. 1972; A simple method for the large-scale preparation of sucrose gradients. FEBS Letters 20:117–119
    [Google Scholar]
  3. Bayley S.A., Duggleby C.J., Worsey M.J., Williams P.A., Hardy K.G., Broda P. 1977; Two modes of loss of the TOL function from Pseudomonas putida mt-2. Molecular and General Genetics 154:203–204
    [Google Scholar]
  4. Benson S., Shapiro J. 1978; TOL is a broad-host-range plasmid. Journal of Bacteriology 135:278–280
    [Google Scholar]
  5. Currier T.C., Nester E.W. 1976; Isolation of covalently closed circular DNA of high molecular weight from bacteria. Analytical Biochemistry 76:431–441
    [Google Scholar]
  6. Downing R., Broda P. 1979; A cleavage map of the TOL plasmid of Pseudomonas putida mt-2. Molecular and General Genetics 177:189–191
    [Google Scholar]
  7. Duggleby C.J., Bayley S.A., Worsey M.J., Williams P.A., Broda P. 1977; Molecular sizes and relationships of TOL plasmids in Pseudomonas. . Journal of Bacteriology 130:1274–1280
    [Google Scholar]
  8. Franklin F.C.H., Williams P.A. 1980; Construction of a partial diploid for the degradative pathway encoded by the TOL plasmid (pWWO) from Pseudomonas putida mt-2: evidence for the positive nature of the regulation by the xylR gene. Molecular and General Genetics 111:321–328
    [Google Scholar]
  9. Freifelder D. 1971; Isolation of extrachromosomal DNA from bacteria. Methods in Enzymology 21:153–163
    [Google Scholar]
  10. Greene P.J., Betlach M.C., Boyer H.W., Goodman H.M. 1974; The EcoRI restriction endonuclease. Methods in Molecular Biology 7:87–111
    [Google Scholar]
  11. Hansen J.B., Olsen R.H. 1978; Isolation of large bacterial plasmids and characterisation of the P2 incompatibility group plasmids pMGl and pMG5 . Journal of Bacteriology 135:227–238
    [Google Scholar]
  12. Hopper D.J., Kemp P.D. 1980; Regulation of enzymes of the 3,5-xylenol-degradative pathway in Pseudomonas putida: evidence for a plasmid. Journal of Bacteriology 142:21–26
    [Google Scholar]
  13. Humphreys G.O., Willshaw G.A., Anderson E.S. 1975; A simple method for the preparation of large quantities of pure plasmid DNA. Biochimica et biophysica acta 383:457–463
    [Google Scholar]
  14. Nakazawa T., Yokota T. 1973; Conjugal transfer of benzoate pathway genes in Pseudomonas arvilla mt 2. apanese Journal of Bacteriology 28:46
    [Google Scholar]
  15. Nakazawa T., Inouye S., Nakazawa A. 1980; Physical and functional mapping of RP4-TOL plasmid recombinants: analysis of insertion and deletion mutants. Journal of Bacteriology 144:222–231
    [Google Scholar]
  16. Palchaudhuri S., Chakrabarty A. 1976; Isolation of plasmid deoxyribonucleic acid from Pseudomonas putida. . Journal of Bacteriology 126:410–416
    [Google Scholar]
  17. Pickup R.W., Wheatcroft R., Williams P.A. 1980; Characterization and analysis of the degradative plasmid DNA in regulatory mutants of Pseudomonas putida MT20. Society for General Microbiology Quarterly 8:43–44
    [Google Scholar]
  18. Richards W.C., Takai S. 1979; A microfuge tube technique for disrupting microquantities of fungal spores. Analytical Biochemistry 100:240–243
    [Google Scholar]
  19. Williams P.A., Murray K. 1974; Metabolism of benzoate and the methylbenzoates by Pseudomonas putida (arvilla) mt-2: evidence for the existence of a TOL plasmid. Journal of Bacteriology 120:4l6–423
    [Google Scholar]
  20. Williams P.A., Worsey M.J. 1976; Ubiquity of plasmids in coding for toluene and xylene metabolism in soil bacteria: evidence for the existence of new TOL plasmids. Journal of Bacteriology 125:818–828
    [Google Scholar]
  21. Worsey M.J., Williams P.A. 1975; Metabolism of toluene and xylenes by Pseudomonas putida {arvilla) mt-2: evidence for a new function of the TOL plasmid. Journal of Bacteriology 124:7–13
    [Google Scholar]
  22. Worsey M.J., Williams P.A. 1977; Characterisation of a spontaneously occurring mutant of the TOL 20 plasmid in Pseudomonas putida MT20: possible regulatory implications. Journal of Bacteriology 130:1149–1158
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-124-2-433
Loading
/content/journal/micro/10.1099/00221287-124-2-433
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error