Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1975 Sep;72(9):3647–3651. doi: 10.1073/pnas.72.9.3647

Transformation of Pseudomonas putida and Escherichia coli with plasmid-linked drug-resistance factor DNA.

A M Chakrabarty, J R Mylroie, D A Friello, J G Vacca
PMCID: PMC433053  PMID: 1103151

Abstract

Conditions optimal for the transformation of Pseudomonas putida and E. coli with a drug-resistance factor (RP 1) DNA, which specifies resistance to carbenicillin, tetracycline, kanamycin, and neomycin, are described. The transformants retain all the fertility, incompatibility, and drug-resistance characteristics present in the parent. Covalently-closed circular molecules of almost identical contour lengths have been isolated from the parent and the transformants. The frequency of transformation is drastically reduced by treatment of RP 1 DNA with DNase and by denaturation or sonication. Shearing of RP 1 DNA in vitro and their subsequent introduction in P. putida cells, by transformation, produces transformants that exhibit a wide range of drug-resistant phenotypes, including those which are resistant to neomycin but sensitive to kanamycin. Isolation of such neomycin-resistant but kanamycin-sensitive transformants indicates that there might be two separate mechanisms specified by RP 1 for resistance to the two antibiotics.

Full text

PDF
3647

Images in this article

3650Image
on p.3650

Selected References

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

  1. Chakrabarty A. M. Genetic basis of the biodegradation of salicylate in Pseudomonas. J Bacteriol. 1972 Nov;112(2):815–823. doi: 10.1128/jb.112.2.815-823.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chang A. C., Cohen S. N. Genome construction between bacterial species in vitro: replication and expression of Staphylococcus plasmid genes in Escherichia coli. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1030–1034. doi: 10.1073/pnas.71.4.1030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cohen S. N., Chang A. C., Boyer H. W., Helling R. B. Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240–3244. doi: 10.1073/pnas.70.11.3240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davies J., Brzezinska M., Benveniste R. The problems of drug-resistant pathogenic bacteria. R factors: biochemical mechanisms of resistance to aminoglycoside antibiotics. Ann N Y Acad Sci. 1971 Jun 11;182:226–233. doi: 10.1111/j.1749-6632.1971.tb30659.x. [DOI] [PubMed] [Google Scholar]
  6. Grinsted J., Saunders J. R., Ingram L. C., Sykes R. B., Richmond M. H. Properties of a R factor which originated in Pseudomonas aeruginosa 1822. J Bacteriol. 1972 May;110(2):529–537. doi: 10.1128/jb.110.2.529-537.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hedges R. W., Jacob A. E. Transposition of ampicillin resistance from RP4 to other replicons. Mol Gen Genet. 1974;132(1):31–40. doi: 10.1007/BF00268228. [DOI] [PubMed] [Google Scholar]
  8. Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ingram L., Sykes R. B., Grinsted J., Saunders J. R., Richmond M. H. A transmissible resistance element from a strain of Pseudomonas aeruginosa containing no detectable extrachromosomal DNA. J Gen Microbiol. 1972 Sep;72(2):269–279. doi: 10.1099/00221287-72-2-269. [DOI] [PubMed] [Google Scholar]
  10. Lindberg M., Novick R. P. Plasmid-specific transformation in Staphylococcus aureus. J Bacteriol. 1973 Jul;115(1):139–145. doi: 10.1128/jb.115.1.139-145.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Morrow J. F., Cohen S. N., Chang A. C., Boyer H. W., Goodman H. M., Helling R. B. Replication and transcription of eukaryotic DNA in Escherichia coli. Proc Natl Acad Sci U S A. 1974 May;71(5):1743–1747. doi: 10.1073/pnas.71.5.1743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Olsen R. H., Shipley P. Host range and properties of the Pseudomonas aeruginosa R factor R1822. J Bacteriol. 1973 Feb;113(2):772–780. doi: 10.1128/jb.113.2.772-780.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Richmond M. H., Sykes R. B. The chromosomal integration of a -lactamase gene derived from the P-type R-factor RP1 in Escherichia coli. Genet Res. 1972 Oct;20(2):231–237. doi: 10.1017/s0016672300013732. [DOI] [PubMed] [Google Scholar]
  14. Shipley P. L., Olsen R. H. Isolation of a nontransmissible antibiotic resistance plasmid by transductional shortening of R factor RP1. J Bacteriol. 1975 Jul;123(1):20–27. doi: 10.1128/jb.123.1.20-27.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Thompson R., Hughes S. G., Broda P. Plasmid identification using specific endonucleases. Mol Gen Genet. 1974;133(2):141–149. doi: 10.1007/BF00264835. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES