Development of a DNA probe for the structural gene of the 2"-O-adenyltransferase aminoglycoside-modifying enzyme.
Analysis of aminoglycoside-resistant Enterobacteriaceae isolated from patients at the Seattle Veterans Administration Medical Center indicated that a single 68-kilobase R factor was responsible for the epidemic spread of low-level resistance to gentamicin, kanamycin, and tobramycin. An examination, by means of the phosphocellulose paper binding assay, of resistant strains carrying this R factor resulted in the identification of a 2"-O-adenyltransferase [ANT(2")]-modifying enzyme. This enzyme was later detected in strains containing 150-kilobase plasmids. For more convenient monitoring of the dissemination of the ANT(2") gene among clinical isolates at the medical center, a DNA probe was developed by cloning of the ANT(2") structural gene from the 68-kilobase factor into pBR322. A 310-base pair Ava I restriction fragment isolated from the interior of the cloned ANT(2") gene was radiolabeled and used in Southern hybridization gels as a probe for plasmids isolated from aminoglycoside-resistant organisms. The probe proved to be highly specific and was more sensitive than enzymologic techniques for detection of the ANT(2") gene in clinical isolates with complex aminoglycoside resistance phenotypes.
D000617:Aminoglycosides / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004274:DNA, Recombinant; D004352:Drug Resistance, Microbial; D004755:Enterobacteriaceae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D009693:Nucleic Acid Hybridization; D009713:Nucleotidyltransferases / Q000235:genetics*; D011815:R Factors
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D009713:Nucleotidyltransferases; C020565:gentamicin 2''-nucleotidyltransferase