Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1976 Sep;127(3):1529–1537. doi: 10.1128/jb.127.3.1529-1537.1976

Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid.

J A Meyers, D Sanchez, L P Elwell, S Falkow
PMCID: PMC232949  PMID: 821935

Abstract

Agarose gel electrophoresis may be employed effectively for the detection and preliminary characterization of plasmid deoxyribonucleic acid (DNA) present in clinical isolates and laboratory strains of gram-negative microorganisms. The method is sensitive and does not require radioisotopes or ultracentrifugation. The estimation of plasmid mass from the extent of DNA migration in gels compares favorably with results obtained by electron microscopy of plasmid DNA purified by equilibrium density centrifugation. The method has proved to be a useful tool for survey work and the epidemiological investigation of plasmid dissemination, as well as an important adjunct to the genetic analysis of plasmids.

Full text

PDF
1532

Images in this article

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. Boyer H. W. Restriction and modification of DNA: enzymes and substrates. Introductory remarks. Fed Proc. 1974 May;33(5):1125–1127. [PubMed] [Google Scholar]
  3. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crosa J. H., Luttropp L. K., Falkow S. Nature of R-factor replication in the presence of chloramphenicol. Proc Natl Acad Sci U S A. 1975 Feb;72(2):654–658. doi: 10.1073/pnas.72.2.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Elwell L. P., De Graaff J., Seibert D., Falkow S. Plasmid-linked ampicillin resistance in haempohilus influenza type b. Infect Immun. 1975 Aug;12(2):404–410. doi: 10.1128/iai.12.2.404-410.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Falkow S., Guerry P., Hedges R. W., Datta N. Polynucleotide sequence relationships among plasmids of the I compatibility complex. J Gen Microbiol. 1974 Nov;85(1):65–76. doi: 10.1099/00221287-85-1-65. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Gyles C., So M., Falkow S. The enterotoxin plasmids of Escherichia coli. J Infect Dis. 1974 Jul;130(1):40–49. doi: 10.1093/infdis/130.1.40. [DOI] [PubMed] [Google Scholar]
  9. Heffron F., Rubens C., Falkow S. Translocation of a plasmid DNA sequence which mediates ampicillin resistance: molecular nature and specificity of insertion. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3623–3627. doi: 10.1073/pnas.72.9.3623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Helinski D. R., Clewell D. B. Circular DNA. Annu Rev Biochem. 1971;40:899–942. doi: 10.1146/annurev.bi.40.070171.004343. [DOI] [PubMed] [Google Scholar]
  11. Humphreys G. O., Willshaw G. A., Anderson E. S. A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta. 1975 Apr 2;383(4):457–463. doi: 10.1016/0005-2787(75)90318-4. [DOI] [PubMed] [Google Scholar]
  12. Mayer L. W., Holmes K. K., Falkow S. Characterization of plasmid deoxyribonucleic acid from Neisseria gonorrhoeae. Infect Immun. 1974 Oct;10(4):712–717. doi: 10.1128/iai.10.4.712-717.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES