Transformation of Pseudomonas putida and Escherichia coli with plasmid-linked drug-resistance factor DNA.
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.
D002118:Calcium / Q000494:pharmacology; D002228:Carbenicillin; D004247:DNA / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism; D004276:DNA, Satellite / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D005111:Extrachromosomal Inheritance; D007612:Kanamycin; D009355:Neomycin; D010403:Penicillin Resistance; D010957:Plasmids; D011549:Pseudomonas / Q000378:metabolism*; D011815:R Factors; D013752:Tetracycline; D014170:Transformation, Genetic / Q000187:drug effects
D004269:DNA, Bacterial; D004276:DNA, Satellite; D007612:Kanamycin; D004247:DNA; D013752:Tetracycline; D002228:Carbenicillin; D009355:Neomycin; D002118:Calcium