Transformation of Pseudomonas putida and Escherichia coli with plasmid-linked drug-resistance factor DNA.
- 1 September 1975
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 72 (9), 3647-3651
- https://doi.org/10.1073/pnas.72.9.3647
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.Keywords
This publication has 10 references indexed in Scilit:
- Plasmid ColE1 as a Molecular Vehicle for Cloning and Amplification of DNAProceedings of the National Academy of Sciences, 1974
- Plasmid identification using specific endonucleasesMolecular Genetics and Genomics, 1974
- Replication and Transcription of Eukaryotic DNA in Esherichia coliProceedings of the National Academy of Sciences, 1974
- Genome Construction Between Bacterial Species In Vitro: Replication and Expression of Staphylococcus Plasmid Genes in Escherichia coliProceedings of the National Academy of Sciences, 1974
- Transposition of ampicillin resistance from RP4 to other repliconsMolecular Genetics and Genomics, 1974
- Construction of Biologically Functional Bacterial Plasmids In VitroProceedings of the National Academy of Sciences, 1973
- The chromosomal integration of a β-lactamase gene derived from the P-type R-factor RP1 inEscherichia coliGenetics Research, 1972
- A Transmissible Resistance Element from a Strain of Pseudomonas aeruginosa Containing No Detectable Extrachromosomal DNAJournal of General Microbiology, 1972
- Nonchromosomal Antibiotic Resistance in Bacteria: Genetic Transformation of Escherichia coli by R-Factor DNAProceedings of the National Academy of Sciences, 1972
- R FACTORS: BIOCHEMICAL MECHANISMS OF RESISTANCE TO AMINOGLYCOSIDE ANTIBIOTICS*Annals of the New York Academy of Sciences, 1971