Construction of a gentamicin resistance gene probe for epidemiological studies.
A 7.7-kilobase BamHI fragment was cloned from the transconjugant of a clinical isolate of Escherichia coli containing a 120-kilobase multiresistance IncC plasmid. The recombinant plasmid conferred resistance to kanamycin, gentamicin, tobramycin, sulfamethoxazole, and trimethoprim. This clone was used to generate a series of subclones from which a 2.0-kilobase BamHI-HindIII probe containing a gentamicin 2''-O-adenylyltransferase [AAD(2'')] gene was obtained. This probe hybridized specifically in both colony and Southern hybridizations with the AAD(2'') gene but not with other resistance genes, including other aminoglycoside-modifying genes, or with a reference IncC plasmid lacking the AAD(2'') gene. The AAD(2'') gene may be part of a transposon, since hybridization occurred with both nonconjugative plasmids and the chromosome in some isolates.
D000617:Aminoglycosides / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D003001:Cloning, Molecular / Q000379:methods; D003428:Cross Infection / Q000382:microbiology*; D004262:DNA Restriction Enzymes; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial / Q000187:drug effects; D005839:Gentamicins / Q000494:pharmacology*; D006801:Humans; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D014176:Protein Biosynthesis
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D005839:Gentamicins; D004262:DNA Restriction Enzymes