The use of a DNA probe and PCR to examine the distribution of the aac(6')-Ic gene in Serratia marcescens and other gram-negative bacteria.
A nucleotide probe for the chromosomal aminoglycoside 6' acetyltransferase gene (aac(6')-Ic) of Serratia marcescens was used in non-radioactive dot-blot hybridization experiments with 186 strains belonging to ten different species of Serratia. The gene was only detected in strains of S. marcescens (all strains tested), and positive hybridization was seen irrespective of whether or not strains were kanamycin-resistant. An additional 180 strains belonging to 28 Gram-negative bacterial species other than in the genus Serratia did not hybridize with the probe. A rapid PCR test for the aac(6')-Ic gene was developed and used to confirm that the aac(6')-Ic gene is only found in S. marcescens. Southern hybridization analysis of S. marcescens chromosomal DNA demonstrated that the gene was usually located on a PvuII fragment with a putative isoleucine tRNA-2 gene, but polymorphisms with respect to the size of this fragment were observed.
Snelling
A M
AM
Department of Microbiology, University of Leeds, UK.
Hawkey
P M
PM
Heritage
J
J
Downey
P
P
Bennett
P M
PM
Holmes
B
B
D000123:Acetyltransferases / Q000235:genetics*; D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001483:Base Sequence; D015139:Blotting, Southern; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D004352:Drug Resistance, Microbial; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012706:Serratia marcescens / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D015342:DNA Probes; D004269:DNA, Bacterial; D000123:Acetyltransferases; C056532:aminoglycoside N(6')-acetyltransferase